Publications

Cells face diverse mechanical stimuli that vary with cell type, state, and pathological conditions. Mechanobiology investigates how cells sense and respond to these forces. While most work has focused on the cell surface and nucleus as primary mechanosensors, how intracellular organelles adapt to extracellular mechanical forces remains largely unknown. Here, we show that extracellular mechanical signals influence the secretory function of the Golgi apparatus. By subjecting adherent cells to mechanical challenges-cell spreading on different ligands, altered substrate stiffness, or equibiaxial strain-we reveal that extracellular forces modulate Golgi-to-cell surface carrier biogenesis, thereby regulating exocytosis. Together with changes in Golgi membrane tension, we identify molecular determinants of the mechanotransduction pathway, including microtubule acetylation, diacylglycerol production, and protein kinase D activity. In turn, inhibition of Golgi export suppresses this mechanoresponse and causes impaired cell spreading. These findings uncover a bidirectional mechanotransduction axis in which extracellular mechanics tune Golgi secretory output, providing a framework for investigating organelle-based mechanoadaptation in physiology and disease.

JTD Keywords: Actin dynamics, Exocytosis, Focal adhesions, Membrane tension, Microtubules, Nucleus, Protein-kinase-d, Secretory protein, Small-molecule inhibitor, Transport

A major challenge in cardiac research is the limited translatability of drug screening and toxicity assays due to the use of in vitro models that poorly mimic the native cardiac environment. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) offer a promising route forward, but conventional 2D culture on rigid substrates hinders their functional maturation and predictive accuracy. This study addresses this problem by investigating the effect of hybrid fibronectin-based hydrogels with tunable stiffness on the mechanical and electrical properties of hiPSC-CMs. We engineered hydrogels with stiffness mimicking the lowest range of neonatal heart tissue stiffness (2-4 kPa) and compared hiPSC-CM behavior on these substrates to that on standard fibronectin-coated glass. Our results demonstrate that hydrogel culture promotes more uniform and stable cardiomyocyte contractions, as evidenced by increased single peak percentages and altered contraction duration. Electrophysiological analysis revealed that hydrogel stiffness influences action potential duration and signal amplitude. Furthermore, hiPSC-CMs on hydrogels exhibited enhanced cell-matrix and cell-cell adhesion, indicating improved structural and functional connectivity. Drug testing with known cardioactive compounds, including isoproterenol and nifedipine, revealed distinct differences in drug responses between hydrogel and glass cultures, suggesting that hydrogels provide a more physiologically relevant platform for assessing drug effects. This work highlights the potential of engineered hydrogel substrates to enhance the functional maturity and predictive accuracy of hiPSC-CMs for cardiac research and drug development.

JTD Keywords: Action potentials, Beat, Cell adhesion, Cell differentiation, Cells, Cells, cultured, Contraction, Fibronectins, Guinea-pig, Humans, Hydrogels, Induced pluripotent stem cells, Ipsc-cardiomyocytes, Ipsc‐cardiomyocytes, Isoproterenol, Maturation, Mechanical properties, Myocytes, cardiac, Nifedipine, Phenotype, Platform, Sensitive dye di-4-anepps

INTRODUCTION Impaired cerebrospinal fluid (CSF) -mediated clearance has been implicated in Alzheimer's disease (AD), but how choroid plexus (ChP) remodeling relates to these processes in vivo remains incompletely understood.METHODS In a large ADNI cohort, we integrated structural magnetic resonance imaging (MRI), amyloid/tau positron emission tomography (PET), and diffusion tensor imaging (DTI) -derived analysis along the perivascular space (ALPS) to characterize ChP changes and their associations with ventricular measures, glymphatic marker, CSF biomarkers, and cognition across diagnostic stages. Our experiments in APP/PS1 mice complemented human analyses.RESULTS ChP volume increased with disease stage, while PET signal decreased. Larger ChP volume was associated with ventricular enlargement, a lower ALPS index, higher cortical amyloid/tau burden, and worse cognitive performance. Glymphatic impairment partially mediated ChP effects on cognition and pathology. APP/PS1 mice recapitulated the key ChP and glymphatic MRI phenotypes observed in humans, alongside altered expression and localization of key proteins.DISCUSSION ChP remodeling linked to impaired CSF-mediated clearance, highlighting its role as an early diagnostic and therapeutic target.

JTD Keywords: Alzheimer disease, Cerebrospinal-fluid, Choroid plexus, Glymphatic system, Magnetic resonance imaging, Mouse, Positron emission tomography, Positron-emission-tomography, Psychoradiology, Tau, Volume

The development of effective antibody therapeutics has been hampered by a lack of methods to measure drug delivery and activity within tumors at single-cell resolution. Here we introduce single-cell spatial pharmacobiology (SSP), an experimental and analytical framework that integrates in situ imaging of a systemically infused, fluorescently labeled therapeutic antibody with high-plex spatial proteomics to quantify antibody distribution, target engagement and tumor microenvironment (TME) architecture. We applied SSP to tumor tissues from participants with head and neck squamous cell carcinoma and pancreatic ductal adenocarcinoma who received the antibody panitumumab-IRDye800 in phase 1 trials. SSP identified pronounced spatial heterogeneity in single-cell drug delivery and target engagement, shaped by conserved stromal barriers, including periostin-rich extracellular matrix assemblies and fibroblast-activation-protein-positive cancer-associated fibroblast neighborhoods, which were associated with reduced antibody delivery in both tumor types. SSP measures drug-target-TME interactions in human tumors and can support studies of resistance mechanisms, dosing strategies and discovery of spatial biomarkers for precision oncology.

JTD Keywords: Cancer, Diffusion, Fibroblasts, Gemcitabine, Head, Inhibitor, Monoclonal-antibodies, Multiplex, Panitumumab-irdye800cw, Transport

Traction force microscopy (TFM) quantifies cellular forces at the cell-extracellular matrix interface, yet elastic formulations neglect viscous dissipation and can misinterpret cellular forces on viscoelastic substrates. We introduce a semi-analytical 2D viscoelastic TFM (veTFM) that generalizes the Boussinesq framework of elastic TFM to Generalized Maxwell (GMX) substrates with one or two components. By combining Fourier and Laplace transforms, veTFM quantifies time-resolved tractions in finite-thickness substrates and resolves stress-free reference and substrate pre-stress. We derive criteria for when elastic regimes remain valid in this framework. This positions veTFM as a scalable extension of standard 2D TFM (eTFM) to viscoelastic substrates, identifying when eTFM remains sufficient, which elastic limit applies, and when full viscoelastic quantification is required. Applied to beating cardiomyocytes, epithelial cells, and dermal fibroblasts cultured on linear-polyacrylamide and alginate viscoelastic hydrogels, veTFM shows that the elastic or viscoelastic regime engaged by the cell depends on timescale matching between the loading rate and the substrate's relaxation times. Notably, for the Generalized Maxwell substrates analyzed here, viscoelastic traction magnitudes scale with the substrate's total dissipation rather than individual relaxation times, with total dissipation setting traction magnitude and timescale matching determining whether the cell engages the substrate in an elastic or viscoelastic regime.

JTD Keywords: Animals, Biomaterials, Cell, Cell-material interactions, Cell–material interactions, Elasticity, Epithelial cells, Extracellular matrix, Extracellular-matrix viscoelasticity, Fibroblasts, Focal adhesions, Guide, Humans, Hydrogels, Microscopy, atomic force, Myocytes, cardiac, Numerical inversion, Semi-analytical methods, Semi‐analytical methods, Stress relaxation, Viscoelastic hydrogels, Viscoelastic traction force microscopy, Viscosity

Non-small cell lung cancer (NSCLC) exhibits disparate responses to anti-angiogenic therapies between its two major histologic subtypes, lung adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC), suggesting a histotype-dependent angiogenesis regulation. Tumor-associated fibroblasts (TAFs) exhibit an activated/myofibroblast-like phenotype in NSCLC, and are emerging as major regulators of tumor progression; yet, their role in controlling angiogenesis in NSCLC remains undefined. Here we analyzed angiogenesis/hypoxia markers in NSCLC, and combined transcriptomics (bulk RNA-seq, scRNA-seq), angiogenesis arrays, genetic perturbations and functional in vitro and in vivo assays to dissect the histotype-dependent production of pro-angiogenic factors in TAFs. We observed greater angiogenesis and reduced necrosis/hypoxia in LUAD compared to LUSC across multiple patient cohorts. The LUAD-TAF secretome was primed for angiogenesis through SMAD3-dependent overproduction of key regulators, particularly TIMP-1 and VEGF-A. We also uncovered a novel function for TIMP-1 in promoting endothelial cell hyperbranching over basal VEGF signaling. Conversely, LUSC-TAFs displayed diminished angiogenic effects despite upregulating HIF-1 alpha and a hypoxia-associated transcriptional signature, owing to their lower SMAD3 and compensatory increase in SMAD2. Our study unveils the critical influence of TAFs in shaping the distinct angiogenic landscapes in LUAD and LUSC through the opposing SMAD2/3 regulation of TIMP-1, VEGF-A and hypoxia signaling. These results also highlight the therapeutic potential of targeting stromal SMAD3/TIMP-1 in LUAD or microenvironmental stressors such as hypoxia and acidosis in LUSC. In addition, these findings provide a biological framework for understanding the histotype-dependent patterns of dissemination, immune evasion, and response to anti-angiogenic therapies in NSCLC.

JTD Keywords: Activation, Adenocarcinoma, Cells, Expression, Fibroblasts, Growth, Metastasis, Nintedanib, Prognosis, Tissue inhibitor

Biomaterial-associated infections pose a significant challenge, impairing tissue integration and frequently leading to implant failure and revision surgeries. Upon implantation, host cells and bacteria compete for colonizing the implant, in a process known as the "race for the surface," which is critical for the long-term survival of the implant. However, conventional biomaterials commonly fail to simultaneously promote cellular integration and prevent infections. To address this issue, we developed a protease-degradable PEG hydrogel functionalized with the RGD integrin-binding motif to enhance cell adhesion and the antimicrobial peptide hLf1-11 (LF) to provide antibacterial activity. This hydrogel was crosslinked using a protease-sensitive peptide (VPM), enabling enzymatic degradation, and dynamic adaptation to the cellular microenvironment (PEG-RGDLF). Non-bioactive but degradable (PEG-50) and neither bioactive nor degradable (PEG-0) hydrogels were included as controls. PEG-RGDLF hydrogels showed an adequate internal structure, with well-defined porosity, swelling capacity, and protease-mediated degradation rate. PEG-RGDLF improved the adhesion, spreading, proliferation, and ALP activity of human bone marrow mesenchymal stem cells (hBMSCs) and reduced the viability and adhesion of Grampositive and Gram-negative bacteria, significantly affecting their morphology. Furthermore, co-culture models were established under two clinically relevant scenarios: "pre-infection" and "post-infection". In both settings, PEG-RGDLF hydrogels supported enhanced cellular responses, with hBMSCs displaying an elongated morphology and improved adhesion. In summary, by integrating cell-instructive and antibacterial properties with a controlled degradation mechanism, this multifunctional hydrogel presents a robust platform for implant-based therapies, actively promoting tissue regeneration while preventing infection, thus addressing the persistent challenge of implant-associated infections in regenerative medicine and clinical applications.

JTD Keywords: Antibacterial peptide, Bacterial adhesion, Bone, Cell adhesion, Cells, Lf, Multifunctional hydrogel, Peg hydrogels, Peptide, Protease-degradable hydrogels, Rgd, Surface, Titanium

The accumulation of microplastics (& micro;Pls) in ecosystems has become a serious environmental concern. Despite increasing awareness, the development of efficient technologies for their removal and remediation remains a major challenge. Herein, we investigate the photocatalytic degradation of polypropylene (PP) & micro;Pls using nanocomposite catalysts composed of permanently polarized hydroxyapatite microparticles (pp-HAp) and rutile TiO2 nanoparticles (TiO2 NPs). pp-HAp, a macroporous bioceramic, is known for its ability to convert gases (CO2 or/and N2) into small value-added chemical products. While pp-HAp serves both as the substrate and thermocatalytic part of the composite, TiO2 NPs are incorporated as the photocatalytic component. A series of XTiO2/ pp-HAp photocatalyts, where X represents the weight percentage of TiO2 (X = 2.5, 5, 10 and 20), were prepared applying a thermal stimulated polarization treatment. The resulting materials show no formation of undesirable calcium titanate. After thorough chemical and physical characterization, the photocatalytic performance of the XTiO2/pp-HAp in the degradation of PP-& micro;Pls was evaluated. Under 1 sun irradiation, the process is selective toward ethanol, which indicates the cleavage of consecutive C-C bonds along the PP chains, with the highest yield obtained for 2.5TiO2/pp-HAp. When the irradiation intensity decreases (0.5 sun), ethanol and isopropanol are produced, suggesting a reduction in the efficiency of the C-C cleavage. In all cases, the nanocomposite catalysts exhibit significantly higher yields than TiO2 NPs or pp-HAp individually, which indicates cooperative electronic effects between both components. Regarding selectivity, in addition to the solar irradiation, the TiO2: pp-HAp ratio also determines which product is obtained. Within the framework of circular economy, our strategy yields added-value chemical products from PP waste using only sunlight and an environmentally friendly catalyst.

JTD Keywords: Anatase, Heterogeneous catalysis, Microplastics, Microplastics removal, Nanocomposites, Photocatalysts, Polypropylene photodegradation, Water treatment plants

Cells sense mechanical forces and convert them into biochemical signals - a phenomenon called mechanotransduction, which regulates fundamental processes in health and disease. Cells can also be engineered to have exogenous, artificial mechanotransduction mechanisms; such systems implement a synthetic form of mechanotransduction, which could be applied to revert malignant phenotypes, modulate immunotherapies or to develop diagnostic tools. Here we review the status of the nascent field of synthetic mechanotransduction. First, we summarize the types of molecular mechanism known to be involved in endogenous mechanotransduction. Then, we explain how, by taking inspiration from endogenous systems, synthetic mechanotransduction systems have been developed. In its simpler form, these systems involve the expression of synthetic genes responding to natural force-sensing (mechanosensitive) proteins. In a more advanced form, they also include synthetic mechanosensitive proteins. Additionally, other systems have been developed to control force generation itself, resulting in control of tissue shape and function. Finally, we review general considerations for the design of synthetic mechanotransduction systems, and future challenges and opportunities.

JTD Keywords: Activation, Cell-adhesion, Fluid shear-stress, Force transmission, Integrin, Mechanical-stress, Membrane tension, Notch, Nuclear-envelope, Optogenetic control

Growth factor mimetics offer great potential for osteogenic biomaterials; yet, their use remains limited, likely due to an incomplete understanding of the effects of the microenvironment on their activity. The extracellular matrices (ECMs) where growth factors are presented in vivo are viscoelastic environments, where dynamic receptor-ligand interactions drive cellular responses. Here, supported lipid bilayers of varying viscosity are used as 2D dynamic ECM models, where the bone morphogenetic 2 (BMP-2) mimetic DWIVA is presented to mesenchymal stem cells alongside the adhesive peptide RGD. DWIVA is demonstrated to have no impact on mechanotransductive processes, including actin organisation, focal adhesion formation and YAP localisation, which are exclusively controlled by viscosity via RGD. Interestingly, DWIVA promotes osteogenic markers' expression only on a viscous bilayer, through a process that involves non-canonical BMP-2 pathways; on a mobile bilayer or on a static control, it lacks osteogenic activity. Crucially, osteogenesis is accompanied by a translocation of BMP receptor 1a to the cell edge, where it colocalises with focal adhesions. Our ECM models hence reveal that both a viscosity-enabled threshold of cell-generated forces and a dynamic environment are necessary to harness the osteogenic potential of DWIVA, uncovering key microenvironment properties for the design of DWIVA-based biomaterials.

JTD Keywords: Activated protein-kinase, Adsorption, Bmp-2, Bone, Cells, Differentiation, Dwiva, Dynamics, Lipid-bilayers, Mesenchymal stem cell, Microscopy, Osteogenesis, Receptor, Viscosity

Constitutive integrin endocytosis and recycling control cell movement and morphology. In contrast, the role of newly synthesized integrins delivered via the biosynthetic pathway has been largely overlooked. We used the retention using selective hooks system to monitor the localization of new integrins exiting the endoplasmic reticulum in space and time. We discovered that new integrin delivery to the plasma membrane is polarized and enhances cell protrusion and focal adhesion growth in an extracellular matrix-ligand-dependent manner. Motor-clutch modeling explained the increased adhesion as higher integrin availability driving recruitment of additional receptors. Unexpectedly, live-cell imaging revealed a small subset of fast-emerging integrin vesicles rapidly transported to the cell surface to facilitate localized spreading. This unconventional secretion depended on cell adhesion and correlated with increased surface levels of immature, high-mannose glycosylated integrin, indicating bypass of the canonical Golgi-dependent secretory pathway. Thus, spatial plasma membrane-targeting of new integrins rapidly alters adhesion receptor availability, providing cells with added plasticity to respond to their environment.

JTD Keywords: Adhesion dynamics, Alpha-5-beta-1 integrin, Endocytosis, Essential roles, Migration, Molecular-dynamics, Talin, Trafficking, Transmission, Unconventional secretion

This study investigates a novel strategy combining ultrashort pulsed-direct laser interference patterning (USPDLIP) and chemical etching to create hierarchically micro- and nanorough topographies on zirconia with improved cell-instructive and antibacterial properties. Linear (L3) and grid (G3) micropatterns of 3 mu m periodicity were fabricated via USP-DLIP, and subsequently treated with hydrofluoric acid to introduce an additional homogeneous nanotopography across the patterns. The individual and combined effects of micropatterning and etching on biological responses were evaluated using human mesenchymal stem cells (hMSCs) and two bacterial strains (Pseudomonas aeruginosa and Staphylococcus aureus) in mono- and co-culture settings. Micropatterns primarily guided cell morphology, alignment, and migration, while the introduced nanotopography enhanced focal adhesion formation and modulated cell-surface interactions. Antibacterial effects were found to be topography- and species-specific: micropatterns restricted P. aeruginosa colonization through a bacterial confinement mechanism, whereas etching-induced nanotopography effectively reduced S. aureus adhesion by limiting contact points. In co-culture assays, hMSC survival depended on a complex interplay between cell spreading and bacterial retention, dictated by surface features. Notably, chemical etching improved the antibacterial potential of the patterns against S. aureus, but it did not result in a synergistic improvement on cellular responses. Indeed, among all tested surfaces, the non-etched linear pattern (L3) consistently exhibited the most favorable outcomes -enhancing hMSC adhesion, migration, and osteogenic differentiation and mineralization, while reducing bacterial colonization and supporting cell survival under infection-like co-culture conditions.

JTD Keywords: Antibacterial, Bacterial adhesion, Biofilm formation, Chemical etching, Dental implants, Differentiation, Energy, Growth, Laser patterning, Migration, Nanotopography, Osseointegration, Osteointegration, Titanium, Topography, Zirconia

Highlights What are the main findings? center dot The smartphone-based portable monitoring system enabled detection of previously undiagnosed sleep apnea among post-stroke patients undergoing rehabilitation. center dot Greater sleep-disordered respiratory events and nocturnal hypoxemia were associated with worse baseline disability and lower rehabilitation metrics. What are the implications of the main findings? center dot The portable system was easy to use, facilitating sleep apnea detection after stroke and supporting broader implementation in rehabilitation settings. center dot Routine screening for sleep-disordered breathing at admission may enable earlier diagnosis and management in patients with substantial hypoxemia/event burden that could slow functional recovery. Sleep-disordered breathing (SDB) is common after stroke and may negatively influence recovery, yet it is frequently underdiagnosed. Portable respiratory monitoring devices could facilitate early SDB screening in these patients. We estimated the prevalence of sleep apnea (SA) using a smartphone-based monitoring system in post-stroke patients and examined associations between respiratory indices, stroke severity and disability (NIHSS, mRS), and rehabilitation outcomes (motor and cognitive Functional Independence Measure; FIM). Consecutive patients admitted to inpatient rehabilitation within three months after a stroke underwent an overnight assessment with a smartphone-based respiratory monitoring device, which estimated the apnea-hypopnea index (AHI), mean and minimum SpO(2), time with SpO(2) < 94% and = 3% and >= 4%). Of the 104 screened patients, 59 were recruited, while 56 had valid recordings. Most patients (89%) had previously undiagnosed SA: 11% mild (AHI >= 5 and = 15 and = 30). Greater event burden and nocturnal hypoxemia were associated with older age, worse baseline disability (mRS), lower admission motor FIMs, and poorer rehabilitation metrics. Smartphone-based portable monitoring is an accessible, easy-to-use approach that may enable earlier identification of SA, particularly in individuals with substantial hypoxemia or respiratory event burden.

JTD Keywords: Apnea, Disturbance, Functional outcomes, Impact, Nocturnal hypoxemia, Portable device, Reliability, Risk, Sleep apnea, Sleep-disordered breathing, Stroke

Additive manufacturing enhances the catalyst performance via hierarchical design. To address environmental and resource concerns, this work aims to fabricate directly recycled 3D-printed monoliths using Direct-Ink Writing (DIW) from 100 % recovered cobalt-zirconia powders. Virgin cobalt-zirconia monoliths were firstly fabricated by DIW of 3.0-7.0 wt% Co-enriched hydrogel-based ceramic inks, followed by calcination at 600 degrees C in a single thermal treatment. After testing the catalytic performance of monoliths in ethanol steam reforming, 3Dprinted cobalt-zirconia monoliths were fragmented and subjected to subsequent milling and sieving steps to recover composite cobalt-zirconia powders with the appropriate properties for reuse in DIW. The recovered powders, inks and monoliths were microstructurally, rheologically and catalytically characterized, and then compared to catalysts constituted by virgin materials. The rheology properties of inks for the recycled and virgin monoliths presented an appropriate printability. Furthermore, the catalytic performance of recycled monoliths was close to that exhibited by virgin catalysts. This study demonstrates the feasibility of directly recycling fully 3D-printed catalysts, potentially reducing the environmental impact with a circular production model to enhance sustainability in the catalyst industry.

JTD Keywords: Additive manufacturing, Catalyst ethanol steam reforming, Cobalt catalysts, Direct-ink writing, Efficient, Heterogeneous catalyst, Hydrogen production, Ionically conductive supports, Monoliths, Nanoparticles, Oxidation, Performance, Recycled ceramics, Regeneration, Solid oxide fuel, Zirconia

In this work, a plasma-assisted and solvent-free synthesis of gold (Au) and gold oxide (Au2O3) nanoparticles (NPs) is explored. Accordingly, only the precursor (i.e., HAuCl4 powder) is exposed to the N-2:O-2 plasma gas mixtures, selected to promote its reduction to Au (HAuCl4-to-Au-0) and/or favor its further oxidation to Au2O3. As a result, the production of highly crystalline spherical Au ultra-small NPs (USNPs) of 2.7 +/- 0.7 nm and Au2O3 edged pyramidal-like NPs of 18 +/- 5 nm is obtained in very short times (i.e., 10 s(-1) min), showing excellent stability without the need to add any ligands. To investigate the potential features of the synthesized particles, they are loaded into alginate hydrogels and tested as multi-detection sensor platforms of H2O2 and NADH in front of several interferents. Although both samples present excellent electrochemical sensing activity (as compared to Au NPs synthesized through the Turkevich method), Au2O3 NPs stand out for its high analytical sensibility toward H2O2 (153.1 and 16.8 mu A mm(-1) cm(-2) for single and multiplexing detection, simultaneously); and capacity to measure NADH in real human urine and low detection limits produced by Escherichia coli (7.5 x 10(3) CFU mL(-1)). Herein, a green, easy, and fast method is proposed to produce stable Au USNPs and Au2O3 NPs, showing great potential for the catalytic and biomedical fields.

JTD Keywords: Au usnps, Au2o3 nps, Multiplexing detection, Nadh, Plasma technologies, Sensing biological environments, Xps

Nanostructured surfaces are increasingly used for cell applications due to their enhanced interactions with numerous cell types; yet, their effects on tissues remain unexplored. To address this limitation, we designed vertical silicon nanopillar (Si-NP) arrays with high density, high aspect ratio and submicrometer diameter, as an optimized geometry based on previous cell-nanostructure studies. Using state-of-the-art in vitro and ex vivo assays, we examined adhesion and biocompatibility of biological samples of different origin and level of complexity -human epithelial-like cell lines, Drosophila imaginal discs and patient-derived lung cancer biopsies-laid on Si-NP arrays or unpatterned flat Si surfaces. Our results demonstrated that Si-NP arrays significantly improved cell and tissue adhesion while preventing oxidative damage and early apoptosis. Consistently, focused ion beam-scanning electron microscopy imaging of cells and tissues showed extended horizontal protrusions and limited vertical wrapping around Si-NP, revealing enhanced cell-NP interactions without cell/tissue penetration. In contrast, flat Si surfaces showed poor adhesion, increased apoptosis, and failed to support tumor biopsy attachment. Interaction with Si-NP arrays upregulated reactive oxygen species (ROS), yet mitochondria-associated ROS remained unchanged, and consequently apoptosis was not induced, indicating that the increased ROS arose from non-mitochondrial compartments and did not compromise viability. Notably, Si-NP arrays matched or outperformed biological responses on tissue culture plastic and Transwell-based assays, which are common in vitro and ex vivo substrates, respectively. These findings provide the first demonstration of the biological suitability of Si-NP arrays for tissue applications in research and clinical translation.

JTD Keywords: Activation, Cancer, Cell-adhesion

Percutaneous medical devices - such as prosthetic limbs, dental implants, and subgingival dental restorations commonly fail due to their susceptibility to infection, posing serious morbidity and mortality and a significant economic burden. Current solutions for subgingival dental restoration failure - such as antimicrobial and degradation-resistant materials - remain largely experimental and have yet to achieve widespread clinical translation. Thus, as a next-generation strategy for improving subgingival restoration longevity, we draw inspiration from the tooth, a long-lasting percutaneous organ with robust soft tissue attachment enabled by cellmatrix adhesive hemidesmosomes (HDs). Promoting HD formation around subgingival restorations - mimicking the natural attachment seen in teeth - may extend dental restoration lifespan by fostering a stable mucosal barrier that blocks bacterial access. A family of scalable biologics-free HD instructive light-curable biosealants (HILBs) was engineered for point-of-care coating for subgingival restorations to trigger and guide beneficial pericellular extracellular matrix structural changes at the HILB surface. Material-based strategies for medical device integration with the human body have traditionally focused on controlling surface properties to provoke desired cellular and tissue responses at the bio/non-bio interface. However, this approach overlooks the evolving pericellular matrix that quickly enrobes cells and overrides engineered surface cues. The control of cell-secreted nascent matrix through HILB surface polarity markedly upregulated HD formation. This bioinstruction is dependent on nascent glycoprotein laminin332 secretion and recognized by HD integrins, which demonstrates the potential of biomaterial surface design to guide secreted pericellular matrix with implications for facilitating tissue repair and enhancing subgingival restoration outcomes.

JTD Keywords: Binding, Bioinspiration, Bioinstructive polar surface, Cells, Expression, Hemidesmosomes, Human keratinocytes, Ionomer, Keratinocytes adhesion, Laminin-5 gamma-2 chain, Light-curable biomaterial, Metalloproteinases, Proteins, Self-etch adhesives, Surface-energy

Aiming to transition towards sustainable design processes, plasma doping methods have been investigated as ultra-fast and solvent-free alternatives to chemical doping strategies. Despite their advantages, the current state-of-the-art plasma-doped materials present low doping percentages. Consequently, their acceptance as a replacement to conventional methods is still disfavoured. In this work, we propose a change in the paradigm by presenting a new approach termed high-performance plasma doping (HPPD) capable of intensively doping material lattices. To do so, HPPD exploits the higher number of available sites in vacancy-engineered materials for introducing dopants through non-thermal plasma (NTP) treatment. For this purpose, hydroxyapatite (HAp) is presented as a representative case example of successful HPPD. Thus, HAp disks with OH- lattice vacancies are prepared and treated for short times with NTP. All the HPPD samples are oxygen-doped successfully, displaying conductivity enhancement of up to one order of magnitude. In addition, doping for the entire material bulk is achieved, reaching a doping replacement efficiency of 50%. The proposed mechanism, based on oxygen diffusion through the OH- HAp columns, is corroborated through density functional theory (DFT) calculations. Results reveal the key role of lattice vacancies as charge imbalances, exercising an electronic pull on reactive gas species. Further assessment of the HPPD HAp is done through catalytic CO2 conversion reactions. Thus, the synthesis of C1-C3 products (including ethanol and formic acid, among others) from CO2 under mild conditions (150 degrees C and 6 bar of CO2) is achieved, realizing a total yield of 537.85 +/- 3.40 mu mol gc-1. Finally, the implications of HPPD and its extension towards other materials are discussed and highlighted by performing a state-of-the-art comparison.

JTD Keywords: Co2, Congo red, Conversion, Nanoparticles, Oxide, Photocatalytic degradation, Sites, Surface, Tio2

Transcutaneous devices such as dental implants frequently fail due to infections at their interfaces with epithelial tissues. These infections are facilitated by the lack of integration between the devices and the surrounding soft tissues. This study aims to improve epithelial integration through surface modification of a transcutaneous implant material (polyetheretherketone (PEEK)). The modification involved covalent bonding of collagen via two distinct methods: (1) nonselective binding through any primary amines present on collagen using carbodiimide-based coupling and (2) site-specific binding to the free amine on the N-terminus of collagen molecules. The second approach preserves active sites responsible for interacting with integrins, crucial for epithelial cell adhesion, located near the C terminus. Both conjugation methods resulted in similar amounts of immobilized collagen; yet, surfaces with 2-PCA-based collagen conjugation exhibited 4 times more free amines. This indicates that fewer amines were used for conjugation in these samples, confirming that 2-PCA selectively binds collagen only through the N-terminus amines. Collagen-conjugated surfaces significantly enhanced HaCaT epithelial cell viability and adhesion compared to unmodified PEEK. Furthermore, 2-PCA-based conjugation resulted in a 2-fold increase in beta 4 subunit gene expression of integrin alpha 6 beta 4 (a key epithelial cell adhesion marker), higher integrin beta 4 immunofluorescence (IF) intensity, and over a 30% improvement in cell retention following mechanical detachment, compared to nonselective conjugation. These findings suggest that selective collagen conjugation on PEEK surfaces increases the accessibility of collagen domains responsible for binding with integrin receptors, which in turn improves epithelial cell attachment, offering a promising strategy for reducing infections and enhancing the longevity of transcutaneous devices.

JTD Keywords: Cell-adhesion, Conjugation, Domains, Fibroblasts, Fibronectin, Integrin, Matrix, Protein, Selective protein conjugation, Soft tissue integration, Strategies, Titanium surfaces, Transcutaneous implants

Biological sex significantly influences the prevalence, incidence, and severity of numerous human diseases, yet it remains an underappreciated variable in biomedical research. Although sexually dimorphic genes contribute to sex-specific traits and disease manifestations, their regulation under metabolic stress is poorly understood. To explore sex-specific metabolic adaptations, we analyzed responses to high-fat diet (HFD)-induced obesity in male and female mice, focusing on the regulation of sex-biased genes. Despite similar adiposity, HFD-fed males displayed more severe metabolic impairments than females, highlighting divergent metabolic outcomes. To investigate the basis for these sex-specific differences, we performed whole transcriptomic profiling of liver and white adipose tissue (WAT) at early (2 wk) and late (12 wk) stages of HFD exposure. Our analysis revealed marked sex-specific gene expression changes across multiple categories, particularly pronounced in male WAT after prolonged HFD feeding. Strikingly, genes exhibiting sexual dimorphism under normal conditions were preferentially modulated in both sexes, comprising up to 46% of all HFD-regulated genes. This led to a substantial loss of sex-biased gene expression in both liver and WAT after HFD exposure, correlating with metabolic dysfunction. Male-biased genes associated with cilia function and estrogen response were among the most affected, showing significant downregulation in male WAT under HFD. Our findings provide a novel perspective on how obesity disrupts sex-specific gene expression in key metabolic tissues, particularly targeting sex-biased genes. By revealing that a considerable proportion of sex-biased genes exhibit HFD-regulated modulation, our study highlights the critical role of these genes in maintaining metabolic health. NEW & NOTEWORTHY Biological sex shapes metabolic tissue physiology, largely through sex-biased gene regulation. Our comprehensive transcriptomic analysis reveals that sex-biased genes in liver and white adipose tissue undergo the most significant regulation during obesity-driven metabolic dysfunction, resulting in a loss of their bias. This disruption highlights a previously unrecognized role of sex-biased genes in maintaining metabolic health in both males and females.

JTD Keywords: Er-alpha, Estrogen-receptor, Female, Gender-differences, High-fat-diet, Insulin resistance, Insulin-resistance, Mitochondrial-function, Obesity, Oxidative-metabolism, Sex differences, Sex-differences, Transcriptomics, Type 2 diabetes, White adipose-tissue

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) and Long Covid-19 (LC-19) are complex conditions with no diagnostic markers or consensus on disease progression. Despite extensive research, no in vitro model exists to study skeletal muscle wasting, peripheral weakness, or potential therapies. We developed 3D in vitro skeletal muscle tissues to map muscle adaptations to patient sera over time. Short exposures (48 H) to patient sera led to a significant reduction in muscle contractile strength. Transcriptomic analysis revealed the upregulation of protein translation, glycolytic enzymes, disturbances in calcium homeostasis, hypertrophy, and mitochondrial hyperfusion. Structural analyses confirmed myotube hypertrophy and elevated mitochondrial oxygen consumption In ME/CFS. While muscles initially adapted by increasing glycolysis, prolonged exposure (96-144 H) caused muscle fragility and weakness, with mitochondria fragmenting into a toroidal conformation. We propose that skeletal muscle tissue in ME/CFS and LC-19 progresses through a hypermetabolic state, leading to severe muscular and mitochondrial deterioration. This is the first study to suggest such transient metabolic adaptation.

JTD Keywords: Chronic fatigue syndrome, Long covid-19, Metabolic adaptatio, Skeletal muscle

OBJECTIVES: Invasive mechanical ventilation (IMV) is crucial for acute respiratory distress syndrome (ARDS) management, but mortality remains high. While spontaneous breathing is key to weaning, excessive respiratory effort may injure the lung and diaphragm. Most existing data on respiratory effort during IMV are based on brief periods of observation, potentially underestimating the burden of inappropriate efforts. This study aims to characterize the evolution of respiratory effort over time in ARDS patients and its relation to survival. We hypothesized that nonsurvivors would spend a greater proportion of time in the high-effort range during the active breathing phase compared with survivors. DESIGN, SETTING, AND PATIENTS: In this prospective cohort study, we continuously recorded airway pressure, flow, esophageal, and gastric pressures in ARDS patients on mechanical ventilation during 7 days after the onset of spontaneous breathing. We analyzed physiologic respiratory effort variables, focusing on the proportion of time spent within defined effort ranges, and compared these data between ICU survivors and nonsurvivors. Statistical analysis was conducted using variance weighted methods to account for variability in the number of respiratory cycles analyzed per patient. This study is registered at ClinicalTrials.gov under identifier NCT06490523. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A total of 1,485,405 respiratory cycles were analyzed from 26 ARDS patients (19 survivors, seven nonsurvivors). Nonsurvivors spent significantly more time in high effort (12% vs. 3%; p = 0.006). In contrast, survivors spent more time in the moderate-effort range (50% vs. 5%; p < 0.001). The time spend with high dynamic transpulmonary driving pressure (> 25 cm H2O) was also significantly different between groups (32% survivors vs. 74% nonsurvivors; p = 0.001). CONCLUSIONS: Patients who die of ARDS are more likely to be exposed to high respiratory effort for prolonged periods of time compared with survivors.

JTD Keywords: Acute respiratory distress syndrome, Adult patients, Epidemiology, Esophageal, Esophageal pressure, Evolution, Lung injury, Mechanical ventilation, Mortality, Pressure support ventilation, Pulmonary, Respiratory effort, Transpulmonary pressure

3D-printing has emerged as a leading technology for fabricating personalized scaffolds for bone regeneration. Among the 3D-printing technologies, vat photopolymerization (VP) stands out for its high precision and versatility. It enables the creation of complex, patient-specific scaffolds with advanced pore architectures that enhance mechanical stability and promote cell growth, key factors for effective bone regeneration. This review provides an overview of the advances made in vat photopolymerization printing of calcium phosphates, covering both the fabrication of full ceramic bodies and polymer-calcium phosphate composites. The review examines key aspects of the fabrication process, including slurry composition, architectural design, and printing accuracy, highlighting their impact on the mechanical and biological performance of 3D-printed scaffolds. The need to tailor porosity, pore size, and geometric design to achieve both mechanical integrity and biological functionality is emphasized by a review of data published in the recent literature. This review demonstrates that advanced geometries like Triply Periodic Minimal Surfaces and nature-inspired designs, achievable with exceptional precision by this technology, enhance mechanical and osteogenic performance. In summary, VP's versatility, driven by the diversity of material options, consolidation methods, and precision opens new horizons for scaffold-based bone regeneration.

JTD Keywords: 3d printing, Additive manufacturing, Bioceramic scaffolds, Bone regeneratio, Ceramic scaffolds, Composite scaffolds, Fabrication, Ha scaffolds, Hydroxyapatite, Hydroxyapatite scaffolds, Mechanical-properties, Regeneratio, Scaffold, Stereolithography, Titanium surfaces, Vat polymerization

Cationic coatings on titanium surfaces are a promising approach for dental and biomedical implants due to their low-cost antimicrobial effect and no need for antibiotics. These coatings are applied on hydroxylated (-OH) surfaces using silanes, such as 3-aminopropyltriethoxysilane (APTES). However, it is unclear whether the concentration of this organofunctional compound affects surface charge or potential toxicity. This study investigated how different concentrations of APTES in cationic coatings on titanium samples influence electrostatic behavior and interactions with bacteria and mesenchymal stem cells (MSCs). Titanium discs served as controls (Ti group) and were first treated by plasma electrolytic oxidation (PEO) to generate -OH groups (PEO group). Subsequently, APTES was applied at 83.8, 167.6, and 251.4 mM, forming PEO+APTES0.3, PEO+APTES0.6, and PEO+APTES0.9 groups, respectively. Surfaces were characterized by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), contact angle, Zeta potential, and profilometry. Microbiological assays assessed initial bacterial adhesion (1 h) and biofilm formation (24 h) using Staphylococcus aureus and Escherichia coli. Cell metabolism was assessed on days 1, 3, and 8, while cell viability was assessed on days 1 and 3 using mesenchymal stem cells. PEO-treated surfaces showed porous morphology, and silanization increased roughness and shifted surfaces toward hydrophobicity. Amines and surface charge changes were confirmed by XPS and Zeta potential. Increasing APTES concentration did not proportionally increase cation number. Crystalline hydroxyapatite oxides were identified following the electrochemical process. SEM, EDS, and FTIR confirmed treatment stability after 28 days of immersion, while tribological tests indicated improved performance for PEO-treated groups. Cationic coatings reduced bacterial adhesion by up to 65%, decreased biofilm Log10 values, and increased dead bacteria proportion. Biocompatibility was confirmed by metabolism and cell viability tests, with the group with lower APTES concentration showing the best performance on day 8, with an 80% higher cell metabolism than day 1. On the other hand, higher concentrations of APTES resulted in reduced cell metabolism. These findings indicate, for the first time, that APTES concentration does not affect electrostatic properties but that lower concentrations are required for cytocompatible cationic coatings.

JTD Keywords: Adhesion, Biofilms, Cationic coating, Cell proliferation, Coatings, Electrostatic interactions, Implant surfaces, In-vitro, Nanoparticles, Osteoblasts, Oxidation, Self-assembled monolayers, Thin-films, Titanium, Wettability

Electron transfer (ET) between redox proteins is an essential process in the respiratory and photosynthetic transport chains. While intra-protein ET is well characterized, the experimental methods to investigate inter-protein ET are limited by the presence of the solvent and by the transient nature of the protein-protein interaction and ET event, which are averaged in protein ensembles. Wiring precisely oriented redox protein partners to the nanoscale electrodes of an electrochemical scanning tunneling microscope allows recording the time- and distance-dependence of the current flowing between them. These methods have revealed that the current flowing between individual protein pairs extends beyond tunneling distances and that it is electrochemically gated. However, the corresponding mechanism and the identity of the charge carriers in aqueous solution remain to be elucidated. To determine the species involved in long-distance charge transport between the redox partner proteins Cc and Cc 1 of the respiratory chain, recordings are performed as a function of pH, in heavy water solutions, and in degassed solutions. It is observed that the spatial span and electrochemical gating of long-distance currents are reduced at high pH, in heavy water, and at low oxygen concentration, showing that the currents are assisted by superoxide anions and by protons.

JTD Keywords: Catalysis, Coupled electron-transfer, Dynamics, Electrochemical stm, Gouy-chapman conduit, Grotthuss (grothuss) proton hopping conduction, Interface, Kinetic isotope effect kie, Mechanism, Mitochondria, Pathways, Ph, Proteins, Proton coupled electron transfer pcet, Reactive oxygen species ros, Reductase, Superoxide radical anion sox, Tyrosine phosphorylation

The electrochemical response of alpha/beta-peptides with (L-Ala-beta-Fpg)n sequence, where beta-Fpg refers to syn 3-amino-2-(2-fluorophenyl)-3-phenylpropanoic acid, has been investigated examining the effects of the peptide length (n = 1-3), the stereochemistry of the beta 2,3-diaryl-amino acid and their self-assembly. alpha/beta-Peptides have been deposited by drop-casting on a conducting polymer (CP) film, which is previously electropolymerized on a stainless steel conducting substrate. The current-potential response of the CP coated by the different studied peptides suggests that, for alpha/beta-peptides, the role played by the electron transport through intermolecular stacking of aromatic side groups prevails over peptide length and stereochemistry. In order to prove such a hypothesis, the experimental conditions used to achieve an ordered self-assembly are optimized for one of the alpha/beta-peptides. The achieved self-assembled structures, which consist of well-defined long microfibers, considerably improve the electrochemical response of the CP. Finally, the prepared alpha/beta-peptide-based electrodes are used to electrochemically detect the oxidation of nicotinamide adenine dinucleotide (NADH). The analytical parameters are better for electrodes with well-defined peptide microfibers than for uncoated CP, corroborating the importance of pi-pi stacking interactions in the response of alpha/beta-peptides.

JTD Keywords: Acid, Alpha/beta-peptides, Beta-peptides, Bioelectronics, Design, Electron-transfer, Foldamers, Nadh, Nadh detection, Nanomaterials, Self-assembly, Stacking interactions

Non-small cell lung cancer (NSCLC) is the most common lung cancer type and one of the deadliest neoplasias worldwide. NSCLC is histologically classified into adenocarcinoma, squamous cell carcinoma, and other less frequent subtypes. Both subtypes and other solid tumors are increasingly regarded as abnormal organs, highlighting the critical role of the desmoplastic tumor stroma rich in cancer-associated fibroblasts (CAFs) in driving tumor progression and therapeutic resistance. This tumor stroma resembles a chronic fibrotic wound and is largely formed by activated/myofibroblast-like alpha-SMA+ CAFs (myCAFs), which are strongly associated with immunosuppression and poor prognosis. Despite the dominance of the myCAF phenotype, we reported a decade ago phenotypic alterations in NSCLC with a strong dependence on the histologic subtype. Subsequent studies using functional assays, single-cell techniques, and in vivo models have refined these initial observations, enhancing our understanding of the biology of both normal fibroblasts/myofibroblasts and CAFs in NSCLC and other cancer types, including their origins, subclassification, and physiopathologic functions. Notably, increasing evidence supports that CAFs can exhibit tumor-restraining or tumor-promoting effects, and current therapeutic efforts aim to shift the balance towards tumor-restraining phenotypes. Here, we review major advances in our understanding of tumor stromagenesis and CAF heterogeneity in both primary tumors and metastasis, including emerging consensus, with a special focus on NSCLC and its frequent dissemination to the brain. We also highlight the critical role of smoking through epigenetic reprogramming of the TGF-beta/SMAD3 pathway. These advances are beginning to delineate how CAF heterogeneity depends on the stage and histologic subtype in NSCLC.

JTD Keywords: Accumulation, Activation, Cancer-associated fibroblasts, Carcinoma-associated-fibroblasts, Desmoplasia, Differentiation, Generation, Localization, Metastasis, Microenvironment, Myofibroblast, Myofibroblasts, Non-small cell lung cancer, Progression, Stroma, Stromagenesis

Medical devices such as vascular grafts, stents, and catheters are crucial for patient treatment but often suffer suboptimal integration with host tissues due to the nature of their surfaces. The materials commonly used, including metals and synthetic polymers, frequently lead to undesired immune responses and device failure. In this context, coating their surfaces with designer proteins has arisen as a promising strategy to improve the device's biointegration. Here, we present a bioinspired method for coating biomaterial surfaces with protein-engineered polymers designed to mimic tailored functions from the native extracellular matrix (ECM). Combining mussel-inspired catechol chemistry with bioorthogonal click chemistry, we developed a modular grafting method for the surface functionalization of metallic and polymeric implants using a bifunctional peptide containing azide and DOPA (3,4-dihydroxyphenylalanine) groups. This simple dip-coating process enabled the fabrication of bioactive elastin-like coatings with precise peptide presentation. The results reveal enhanced bioactivity and cytocompatibility, as evidenced by improved endothelial cell adhesion, proliferation, and heparin-binding capacity on coated surfaces. The versatility and effectiveness of this bioorthogonal coating method suggest significant potential for creating implant surfaces tailored to diverse clinical applications.

JTD Keywords: Adhesion, Binding, Biofunctional coatings, Cell response, Click chemistry, Coatings, Dopa, Elastin-like recombinamers, Fibronectin, Peptides, Polymers, Protein adsorption, Surface functionalization, Surfaces, Titanium

Unjamming transitions from a solid-like to a fluid-like state are a gateway to breast epithelial cancer invasion. However, the mechanical interplay between phase transitions and dimension transitions, in particular wetting, remains elusive, despite being critical for understanding the onset of metastatic dissemination. This study shows that unjamming, mediated by the RAB5A GTPase, alters carcinoma spheroid fluidity, rigidity, and rewires adhesion mechanics to drive supracellular active wetting as a new mode of tumor expansion. Spheroid fluidification enhances the selective expression of integrin subunits and increases focal adhesion dynamics, inducing a fluid-like spreading behavior on specific matrix ligands. Notably, nanoscale regulation of integrin clustering can select for distinct phase transitions at the collective scale upon wetting. In this framework, fluidized spheroids polarize into cohesive "supracells", and maintain a stiff peripheral actin bundle as measured by nanomechanical mapping. Furthermore, a combination of Brillouin microscopy and 2.5D traction force analysis reveals a mechanical switch within the spheroid core, characterized by significant cell softening and a reduction in compressive forces exerted on the substrate, thereby mimicking the wetting of a liquid droplet. These findings establish unjamming-driven active wetting as a key mechanism to comprehend the molecular and biophysical underpinnings of solid tumor invasion.

JTD Keywords: Adhesion, Biophysics, Dynamics, Epithelia, Integrin, Mechanobiology, Mechanotransduction, Mesenchymal transitions, Paxillin, Rab5a-mediated breast carcinoma fluidification, Snai, Softening, Spheroid wetting, Supracellular motilit

The movement of cells and microorganisms in response to chemical gradients, chemotaxis, is fundamental to the evolution of myriad biological processes. In this work, we demonstrate that even the simplest cell-like structures are capable of chemotactic navigation. By encapsulating enzymes within lipid vesicles that incorporate a minimal number of membrane pores, we reveal that a solitary vesicle can actively propel itself toward an enzyme substrate gradient. Specifically, vesicles loaded with either glucose oxidase or urease and embedded with corresponding transmembrane proteins were tracked within a microfluidic device under a controlled substrate gradient. Our findings establish that a system comprising only an encapsulated enzyme and a single transmembrane pore is sufficient to initiate chemotaxis. This proof-of-concept model underscores the minimalistic yet powerful nature of cellular navigation mechanisms, providing a previously unknown perspective on the origins and evolution of chemotactic behavior in biological systems.

JTD Keywords: Alpha-hemolysin, Artificial cells, Colloid transport, Gradients, Migration, Model, Phospholipid-vesicles, Propulsion

When exposed to the oral environment, dental implants, like natural surfaces, become substrates for microbial adhesion and accumulation, often leading to implant-related infections-one of the main causes of implant failure. These failures impose significant costs on patients, clinicians, and healthcare systems. Despite extensive research, there is no consensus on the most effective protocol for managing peri-implantitis. Biomedical engineering has aimed to address this challenge by developing biocompatible implants with surface properties designed to enhance biological responses and reduce polymicrobial accumulation. Due to the complexity of interactions between implants and biological systems, no single material property can drive these processes. Instead, a combination of physical, chemical, and mechanical properties is required to ensure a safe and effective response. Antimicrobial coatings are developed either by incorporating antimicrobial agents onto surfaces or modifying the material's physicochemical properties. These coatings utilize a range of compounds for contact-killing or as drug-delivery systems. While biomaterials science has advanced rapidly in enhancing implant surfaces, these bioengineering techniques have progressed more rapidly than our understanding of the pathogenesis of implant infections. To bridge this gap, biomedical engineering must address emerging knowledge about implant infections, focusing on controlling microbial accumulation while simultaneously managing inflammatory responses to support tissue healing. This review critically evaluates current evidence on implant infection pathogenesis, antimicrobial coating technologies, and systematically assesses their in vivo (animal and human evidence) efficacy to guide future advancements in implant infection mitigation.

JTD Keywords: Antimicrobial, Bacterial adhesion, Biofilm formation, Biomaterials, Coating, Dental implant, Drug-deliver, Free-energy, In-vivo antibacterial, Infectio, Infection, Peri-implantitis, Pure titanium, Quality-of-life, Resistant staphylococcus-aureus, Titanium implant

Surface topography at the nanoscale plays a crucial role in modulating the biological properties of dental implants. However, the understanding of how the nanoroughness of zirconia affects cell and bacteria responses remains unclear. In this study, chemical etching of 3Y-TZP was explored to develop a nanotopography capable of favoring eukaryotic cell behavior while simultaneously inhibiting bacterial adhesion. Three topographies of different roughness were created by varying the etching time with hydrofluoric acid (i.e., HF15, HF30, and HF60). The etched surfaces exhibited a nanorough topography with randomly distributed nanopits, and surface roughness increased at longer etching times. Mesenchymal stem cell adhesion, spreading, proliferation and mineralization were enhanced on the etched surfaces, compared to flat controls. The roughest surface (HF60) also inhibited S. aureus adhesion and caused significant damage to P. aeruginosa. This study highlights the potential of chemical etching to produce nanorough zirconia with improved biological outcomes.

JTD Keywords: Attachment, Bacteri, Bacterial adhesion, Biomaterials, Cell response, Chemical etching, Dental implants, Dental zirconia, Integration, Osseointegration, Osteogenic differentiation, Parameter, Roughness, Surface-topography, Titanium implants, Zirconia

Bacterial colonization on biomaterials is a major issue, leading to approximately 20% of implant failures due to infection and biofilm formation. To address this, peptide-based hydrogels incorporating tailored bioactive peptides have emerged as promising candidates for applications in tissue engineering and infection control. Here, we have designed peptide sequences that incorporate i) a self-assembling unit (SaU) and ii) bioactive motifs, including the well-known arginine-glycine-aspartate (RGD) sequence to promote cell adhesion or an antimicrobial peptide derived from lactoferrin (LF) to exhibit antibacterial properties. To aid in the gelation, these peptides were combined with hyaluronic acid (HA), rendering peptide-based hydrogels without the need for additional external assembly triggers, simplifying their application in biomedical contexts. This protocol allowed for a spontaneous formation of a 3D fibrillar network, with structural and physicochemical properties suitable for tissue engineering. The biological evaluation revealed the ability of RGD-based hydrogels to increase the adhesion and spreading of osteoblastic cells compared to controls, while the LF-based hydrogels significantly reduced the viability and attachment of both Gram-positive and Gram-negative strains, clearly affecting bacterial morphology. This report demonstrates the feasibility of this technology to produce hydrogels incorporating distinct biological cues, highlighting their potential as versatile biomaterials to address diverse biomedical challenges.

JTD Keywords: Alginate, Antimicrobial peptide, Bacterial adhesion, Biomaterial, Cell adhesion, Circular-dichroism, Design, Hlf1-11 peptide, Human lactoferrin, Hyaluronic-acid, Lf, Molecules, Peptide-based hydrogels, Rgd, Self-assembling peptide, Titanium

Gradients are widespread in nature, including within the human body, making the study of nanomotors' collective dynamics in gradients crucial to advancing biomedical applications and deepening the understanding of natural active matters. However, the comprehensive understanding of nanomotors' collective dynamics under gradients remains underexplored, particularly. This study employs urease-based nanomotors (UrNMs) as a model system to explore their collective dynamics within a urea gradient, revealing three fundamental principles that govern their behavior: density-driven convection, UrNMs' response to the urea gradient, and a coupling effect between UrNMs and their environment. Initially, migration is dominated by convection-induced motion arising from the steep gradient. As convection gradually diminishes, UrNMs' positive response to the urea gradient becomes the dominant factor governing their migration. Notably, the coupling effect between nanomotors and the gel, plays a crucial role in the migration process. This coupling effect arises from hydrogen bonding between product anions and the gel, which generates ionic gradients. The dominant influence of electric force is validated by pH-controlled experiments. These insights advance the fundamental understanding of gradient-responsive nanomotor behavior and offer inspiration for the design of intelligent, environment-sensitive active systems.

JTD Keywords: Chemotaxis, Collective behavior, Gradient, Nanomotor

Skin is a barrier that protects us against physical, chemical and biological agents. However, any damage to the skin can disrupt this barrier and therefore compromise its function leading to sometimes catastrophic consequences like sepsis. Thus, methods to detect early signs of infection are necessary. In this work, we have developed a straightforward method for producing 2D nanomembranes with regularly spaced 1D metallic nanostructures integrating sensing capabilities to pH and NADH (nicotinamide adenine dinucleotide), which are critical analytes revealing infection. To achieve this, we have successfully fabricated a bilayered nanomembrane combining a pH-responsive polyaniline (PANI) layer and a nanoperforated poly(lactic acid) (PLA) layer containing gold nanowires (Au NWs) as NADH sensing element. SEM, FTIR, Raman and AFM techniques revealed the formation of the bilayered PANI/PLA nanomembrane and the successful incorporation of the Au NWs inside the nanoperforations. The resulting bilayered nanomembrane showed significant flexibility and conformability onto different substrates due to the softness of the polymers and the ultrathin thickness with stiffness values similar to human skin. These nanomembranes also exhibited remarkable electrochemical sensing performance towards pH and NADH detection. Thus, the nanomembrane displayed linearity with good sensitivity (47 mVpH-1) in the critical pH range 4-10 and fast response time (10 s). On the other hand, PANI/PLA-Au nanomembranes also allowed the quantitative sensing of NADH with a limit of detection of 0.39 mM and a sensitivity of 1 mu A cm-2 mM-1 in the concentration range 0-5 mM.

JTD Keywords: Bilayered nanomembranes, Biomimetic membranes, Free-standing films, Gold nanowires, Nad, Nanoparticles, Pani, Ph and nadh sensor, Pla, Polyaniline

Antifouling coatings are vital to enhance the performance of medical devices, aiming to mitigate bodily reactions by shielding their surface. Despite significant advancements in antifouling coatings, like those based on zwitterionic monomers and hydroxyl-functionalized (meth)acrylamides, limitations like decreased antifouling properties after functionalization and complement system activation hinder their application in blood. Here, a novel class of ultrathin surface-attached hydrogels is presented, consisting of hydrophilic non-charged green solvent-based monomers and preventing protein adsorption while offering on-demand degradability. Unlike the best antifouling brushes, the coatings are easily applicable, unaffected by charges, and free of complement system-activating groups. The hydrogels are formed using copolymers of N,N-dimethyl lactamide acrylate (DMLA) and benzophenone acrylate (BPA). Moreover, 5,6-benzo-2-methylene-1,3-dioxepane (BMDO) is incorporated to introduce hydrolyzable ester. The coating of state-of-the-art devices is demonstrated with X-ray photoelectron spectroscopy (XPS), analyze surface energy components, and confirm their antifouling properties with surface plasmon resonance (SPR). The coatings are non-cytotoxic toward MRC-5 fibroblasts, exhibit repellency against methicillin-resistant Staphylococcus aureus (MRSA), and effectively prevent thrombus formation on devices in blood. This work establishes a versatile platform for next-generation coatings in medical and industrial applications, matching the antifouling efficiency of the most advanced solutions and offering regeneration of substrates by erasing the coating.

JTD Keywords: Adhesion, Antifouling, Biocompatibility, Blood-plasma, Coating, Coatings, Contact, Copolymers, Degradability, Energy components, Green solvent, Hemocompatibilit, Hydrogel, Polyethylene oxide, Polymer brushes, Protein adsorption, Thi

Neurodegenerative diseases such as Alzheimer's disease and amyotrophic lateral sclerosis are characterized by progressive neuronal loss and the accumulation of misfolded proteins including amyloid proteins. Current therapeutic options include the use of antibodies for these proteins, but novel chemical strategies need to be developed. The disaccharide trehalose has been widely reported to prevent misfolding and aggregation of proteins, and we therefore investigated the conjugation of this moiety to biocompatible peptide amphiphiles (TPAs) known to undergo supramolecular polymerization. Using X-ray scattering, circular dichroism, and infrared spectroscopy, we found that trehalose conjugation destabilized the internal beta-sheet structures within the TPA supramolecular polymers as evidenced by a lower thermal transition. Thioflavin T fluorescence showed that these metastable TPA nanofibers suppressed A42 aggregation. Interestingly, we found that the suppression involved supramolecular copolymerization of TPA polymers with A beta 42, which effectively trapped the peptides within the filamentous structures. In vitro assays with human induced pluripotent stem cell-derived neurons demonstrated that these TPAs significantly improved neuron survival compared to other conditions. Our study highlights the potential of properly tuned supramolecular polymerizations of monomers to safely remove amyloidogenic proteins in neurodegeneration.

JTD Keywords: Aggregation, Beta, Disease, Model, Molecule, Motor-neurons, Nanoparticles, Nanostructures, Promote, Trehalose

Additive manufacturing technologies are revolutionizing the fabrication of ceramic catalysts through hierarchical design to enhance catalytic performance and simultaneously improving the efficiency of the manufacturing process by decreasing the initial investment and production steps. This work proposes a fabrication process of cobalt-zirconia monoliths based on Direct-Ink Writing of Co-enriched hydrogel-based ceramic inks, and the debinding and sintering at 600 degrees C in a single thermal treatment. The effect of Co precursor amount (3.0 -7.0 wt% Co) on the rheological properties of inks and the catalytic performance in ethanol steam reforming is investigated. The results reveal the successful incorporation of Co into rectilinear monoliths with 50% infill, obtaining strongly Co-rich surfaces. The remarkable catalytic performance of the 5.0 wt% Co monolith at 300-600 degrees C confirms the feasibility of this novel single-step approach, reaching an appropriate balance between catalytic activity and printability. This outcome may represent a push towards the fabrication of fully 3D-printed monolithic catalysts.

JTD Keywords: Additive manufacturing, Catalyst ethanol steam reforming, Cleanup, Co, Combustion, Direct-ink writing, Hydrogen productio, Ionically conductive supports, Nanoparticles, Oxidation, Rama, Reactors, Sulfur, Zirconia

The bone marrow is a complex tissue with distinct cellular and mechanical heterogeneity, serving as the primary site for haematopoiesis. Under certain conditions, such as on the onset of mutations to healthy cells or alterations to the environment, the bone marrow can also become the origin of haematological malignancies, often characterized by uncontrolled self-renewal of hematopoietic stem cells, overproduction of immature progeny and remodelling of the tissue microenvironment. This remodelling alters the composition and mechanics of the extracellular matrix (ECM), facilitating the proliferation and metastasis of leukaemic cells. The elastic and dissipative properties of the ECM are hallmarks of both health and disease progression in different tissues. However, studying the mechanical properties of bone marrow is difficult owing to inaccessibility in situ. Advanced three-dimensional bioengineered models offer a way to recapitulate the mechanical properties of the bone marrow, but it remains challenging to incorporate the elastic and viscous components. Understanding the physiological and disease-specific mechanical ECM signatures is crucial for advancing bone marrow research and for developing therapeutics. In this Review, we explore the structure-function relationship of the bone marrow, emphasizing its complex mechanical behaviour, and discuss the bioengineered models that recapitulate the mechanical properties in the healthy and diseased bone marrow niche, stressing the importance of replicating ECM physiological and pathological mechanical signatures in the future.

JTD Keywords: Acute myeloid-leukemia, Extracellular-matrix stiffness, Fibrosis, Hematopoietic stem-cell, Lysyl-oxidase, Mesenchymal stromal cells, Microenvironment, N-cadherin, Progenitor cells, Suppor

Cells are sensitive to the physical properties of their microenvironment and transduce them into biochemical cues that trigger gene expression and alter cell behavior. Numerous proteins, including integrins, are involved in these mechanotransductive events. Here, a novel role for the boron transporter NaBC1 is identified as a mechanotransducer. It is demonstrated that soluble boron ions activate NaBC1 to enhance cell adhesion and intracellular tension in C2C12 myoblasts seeded on fibronectin-functionalized polyacrylamide (PAAm) hydrogels. Retrograde actin flow and traction forces exerted by these cells are significantly increased in vitro in response to both increased boron concentration and hydrogel stiffness. These effects are fibronectin and NaBC1-mediated as they are abrogated in hydrogels coated with laminin-111 in place of fibronectin and in esiRNA NaBC1-silenced cells. These findings thus demonstrate that NaBC1 controls boron homeostasis and also functions as a mechanosensor.

JTD Keywords: Activation, Animals, Beta-1-integrin, Biomaterials, Boron, Cell adhesion, Cell line, Cell-matrix, Differentiation, Fibronectins, Focal adhesion kinase, Growth, Hydrogels, Integrins, Mechanobiology, Mechanotransduction, Mechanotransduction, cellular, Mediated adhesion, Mice, Muscle cells, Myoblasts, Nabc1, Skeletal-muscle, Stiffnes, Tissue engineerin, Tissue engineering

Drug targeting can be achieved by coupling drugs or their carriers to affinity molecules, mostly antibodies (Abs), which recognise specific protein targets. However, most proteins are not expressed in an exclusive configuration but as various isoforms. Hence, selected targeting molecules may fail to target with enough efficiency in clinical trials, which is overlooked. We illustrate this by targeting intercellular adhesion molecule 1 (ICAM-1), a cell-surface protein overexpressed in many pathologies. Most ICAM-1 targeting studies used Ab R6.5, which binds ICAM-1 domain 2 (D2). Yet, literature and our data show that D2 is frequently absent among ICAM-1 isoforms. We thus produced a battery of five new Abs (B4, B6, B11, C12 and G2) and tested their ability to recognise both full-length and -D2 ICAM-1. In solution, all Abs recognised both ICAM-1 forms (from 5.3 x 1011 to 4.2 x 1012 sum intensity/well). Coating them on nanocarriers (NCs) rendered G2 specific against -D2 ICAM-1 (4.2 x 106 NCs/well) while other Abs kept their dual recognition (from 6.4 x 106 to 2.2 x 107 NCs/well). All Abs induced NC intracellular uptake in respective cells (from 42% to 85%) and displayed good cross-species reactivity (from 4.4 x 1011 to 2.6 x 1012 sum intensity/well). These Abs represent valuable tools to target ICAM-1 and illustrate a new targeting paradigm that may improve classical strategies.

JTD Keywords: Adhesion, Antibody-targeted nanocarriers, Cross-species reactivit, Design, Domai, Endothelial delivery, Enlimomab, Icam-1, Icam-1 isoforms, Intercellular adhesion molecule 1, Nanocarriers, Nanoparticles, New recombinant antibodies, Pecam-1, Targeting and intracellular trafficking

The possibility of using light to image and manipulate neuronal activity, at the heart of Neurophotonics, has provided new irreplaceable tools to study brain function. In particular, the combination of multiphoton microscopy and optogenetics allows researchers to interact with neuronal circuits with single-cell resolution in living brain tissues. However, significant optical challenges remain to empower new discoveries in Neuroscience. This Review focuses on three critical areas for future development: (1) expanding imaging and optogenetic stimulation to larger fields of view and faster acquisition speeds, while maintaining single-cell resolution and minimizing photodamage; (2) enabling access to deeper brain regions to study currently inaccessible neuronal circuits; and (3) developing optical techniques for studying natural behaviors in freely moving animals. For each of these challenges, we review the current state-of-the-art and suggest future directions with the potential to transform the field.

JTD Keywords: 2-photon excitation, Adaptive optics, All-optical brain studies, All-optical electrophysiology, Calcium and voltage imaging, Field-of-view, High-speed, In-vivo, Large-scale, Multiphoton microscopy, Neural activity, Neurophotonics, Optogenetic photostimulation, Optogeneticphotostimulation, Primary visual-cortex, Voltage indicator, Wavefront shaping, Wavefrontshaping

Arborescent (dendrigraft) polymers are high-molecular-weight dendritic macromolecules with a regular, multilevel branched topology and a high density of functional end groups in their periphery. Their well-defined architecture, devoid of cross-links or loops, imparts a particle-macromolecule duality that becomes particularly pronounced at interfaces. However, the underlying mechanisms governing their interfacial behavior remain largely unexplored. Here, we elucidate how the unique topology dictates the interfacial organization of water-soluble arborescent polymers. Using an iterative grafting-from approach via single-electron transfer living radical polymerization, we synthesized narrowly dispersed polymers with controlled branching and ultra-high molecular weight of 6.2 x 106 g mol-1. These polymers transition from spherical rigid particles in solution, to highly flexible, two-dimensional conformations upon interfacial adsorption. At solid interfaces, increasing segment density shifts surface morphologies from quasi-2D discs to fried-egg-like structures, as observed by atomic force microscopy and corroborated by dissipative particle dynamics simulations. At liquid-liquid interfaces, the absence of substrate constraints facilitates complete spreading into uniform 2D discs, driven by the energy gain due to polymer-segment adsorption. Furthermore, we uncover that macromolecular crowding and topological constraints inherent to the arborescent architecture dictate the response to compression of the adsorbed polymer layer, contrasting sharply with the behavior of conventional flexible linear or star polymers. The combination of high interfacial activity, spatially adaptable end groups, and extreme molecular flexibility will enable arborescent polymers to adapt to complex interfaces, acting as versatile platforms for multivalent and superselective interactions. These properties open new avenues for designing multivalent nanocarriers and adaptive interfacial materials with cooperative binding effects.

JTD Keywords: Angle neutron-scattering, Architectur, Graft, Microgels, Polymers, Polystyrene-graft-poly(2-vinylpyridine) copolymers, Polystyrenes, Radical polymerization, Set, Unimolecular micelles

Recent studies have revealed that cardiac tissue regeneration is promoted by administering an initial dose of exogenous lactate and locally maintaining an abundant concentration of this compound for a prolonged period (i.e., around 10-14 days) through sustained release. The aim of this study is to develop a scaffold based on poly(lactic acid) (PLA) for achieving a sustained daily release of lactate from the first day to the end of the recommended period. First, a five-layered electroresponsive scaffold has been engineered using three PLA layers (first, third, and fifth), each composed of electrospun microfibers (MFs), separated by spin coated lactate (second) and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) (fourth) intermediate layers. The hydrophobicity of the outer PLA layers (first and fifth) has been used to maintain the release of lactate from the intermediate second layer over 3 days, while the conducting fourth PEDOT:PSS layer has ensured a complete lactate release by electrostimulation. After that, in a second step, the same scaffold has been re-engineered to maintain the sustained release not only for a short period (3 days) but also for a prolonged period (>10 days). For this purpose, the PLA MFs of the intermediate third layer have been substituted by plasma-treated proteinase K-containing PLA MFs, obtained by electrospinning a PLA:enzyme mixture. The activity of the enzyme, which decomposes the ester bonds of PLA, combined with the effect of the plasma on the PLA structure, results in a prolonged sustained release that, in addition, can be modulated.

JTD Keywords: 4-ethylenedioxythiophene), Cardiac tissue regeneratio, Conducting polymer, Drug-release, Electroresponsivescaffolds, Electrospinning, Energy, Enzymatic degradation, Hydrogel, Nanofibers, Poly(3, Poly(lactic acid), Raman-spectroscopy

Silica-based nanoparticles have been extensively investigated as advanced bioimaging probes and smart nanocarriers for the development of nanomedicine because of their unique properties. The deep understanding of nanoparticle-biological system (nanobiology) interactions is an important determinant for the success of nanomedicine. In this Review, we focus on the assessment of significant nanobio interactions along with the intracellular fates of silica-based nanoparticles by advanced microscopy and link this information to their recent biomedical applications. Initially, the fundamental concepts of the dynamic interactions between nanoparticles and proteins followed by cells and key factors that influence these nanobio interactions are briefly introduced. Furthermore, the basic principles of advanced imaging modalities for the analysis of nanobio interactions in this study are described. Next, the utilization of advanced imaging for the characterization of protein coronas and monitoring the cellular internalization and intracellular trafficking/fate of individual nanoparticles are comprehensively summarized. Finally, recent biomedical applications of nanoparticles in bioimaging and drug delivery are discussed.

JTD Keywords: Adsorption, Cellular uptake, Desig, Drug-delivery, Mesoporous organosilica nanoparticles, Nanocapsules, Protein corona, Size, Superresolution microscopy, Surface-properties

Bacteria often carry multiple genes encoding anti-phage defense systems, clustered in defense islands and phage satellites. Various unrelated anti-phage defense systems target phage-encoded homologous recombinases (HRs) through unclear mechanisms. Here, we show that the phage satellite SaPI2, which does not encode orthodox anti-phage defense systems, provides antiviral immunity mediated by Stl2, the SaPI2-encoded transcriptional repressor. Stl2 targets and inhibits phage-encoded HRs, including Sak and Sak4, two HRs from the Rad52-like and Rad51-like superfamilies. Remarkably, apo Stl2 forms a collar of dimers oligomerizing as closed rings and as filaments, mimicking the quaternary structure of its targets. Stl2 decorates both Sak rings and Sak4 filaments. The oligomerization of Stl2 as a collar of dimers is necessary for its inhibitory activity both in vitro and in vivo. Our results shed light on the mechanisms underlying antiviral immunity against phages carrying divergent HRs.

JTD Keywords: Bacteri, Crystal-structure, Escherichia-coli reca, Gene, Inhibition, Mechanism, Pathogenicity island interference, Protein, Rad52, Sos-response

Extracellular matrix (ECM) viscoelasticity has emerged as a potent regulator of physiological and pathological processes, including cancer progression. Spatial confinement within the ECM is also known to influence cell behavior in these contexts. However, the interplay between matrix viscoelasticity and spatial confinement in driving epithelial cell mechanotransduction is not well understood, as it relies on experiments employing purely elastic hydrogels. This work presents an innovative approach to fabricate and micropattern viscoelastic polyacrylamide hydrogels with independently tuneable Young's modulus and stress relaxation, specifically designed to mimic the mechanical properties observed during breast tumor progression, transitioning from a soft dissipative tissue to a stiff elastic one. Using this platform, this work demonstrates that matrix viscoelasticity differentially modulates breast epithelial cell spreading, adhesion, YAP nuclear import and cell migration, depending on the initial stiffness of the matrix. Furthermore, by imposing spatial confinement through micropatterning, this work demonstrates that confinement alters cellular responses to viscoelasticity, including cell spreading, mechanotransduction and migration. These findings establish ECM viscoelasticity as a key regulator of epithelial cell mechanoresponse and highlight the critical role of spatial confinement in soft, dissipative ECMs, which was a previously unexplored aspect.

JTD Keywords: Cell adhesion, Cell movement, Confinement, Dynamics, Elastic modulus, Elasticity, Epithelial cells, Extracellular matrix, Extracellular-matrix, Force transmission, Humans, Hydrogels, Mechanics, Mechanotransduction, cellular, Micropatterning, Migration, Morphology, Motilit, Polyacrylamide hydrogels, Stiffness, Substrate, Viscoelasticit, Viscoelasticity, Viscosity

Intercellular adhesion molecule-1 (ICAM-1) is a membrane protein whose expression is enhanced at pathological sites, supporting drug delivery using nanocarriers (NCs). Any of its five extracellular domains (D1 to D5) can be targeted, yet most NC studies have used antibody (Ab) R6.5, which targets domain D2. While this provided efficient NC targeting and intracellular transport, literature indicates the absence of D2 in about 50 % of ICAM-1 isoforms expressed in mouse models. In this study, we verified the presence of ICAM-1 isoforms lacking D2 in human cells at both mRNA and protein levels, supporting the need to test Abs targeting other ICAM-1 domains. We developed a new cell model specifically lacking ICAM-1 D2 and compared R6.5 to Abs targeting D1 (Ab 15.2), D3D4 (Ab G-5), and D5 (Ab H-4). Abs G-5 and H-4 showed best targeting results, for which they were coated on model polymeric NCs. Compared to non-specific IgG NCs, both anti-ICAM-1 formulations targeted recombinant cells expressing human ICAM-1 lacking D2 and also primary cells naturally expressing the whole ICAM-1 isoform pattern observed. Both formulations were efficiently internalized by cells and trafficked to lysosomes, as previously observed for ICAM-1-targeting systems. Furthermore, NCs coated with either one of these two Abs showed good cross-species reactivity, being amenable for future pre-clinical testing. Therefore, Abs G-5 or H-4 are good options to provide ICAM-1 targeting without missing ICAM-1 isoforms lacking D2, present in human.

JTD Keywords: Adhesion molecule-1 icam-1, Anti-icam-1 antibody, Antibody-targeted nanocarriers, Design, Different receptor epitopes, Domai, Endothelial delivery, Enlimomab, Icam-1 extracellular domains, Icam-1 isoforms, Identification, Intercellular adhesion molecule 1, Monoclonal-antibodies, Nanoparticles, Targeting and endocytosi, Transport

Multicellularity is one of the major evolutionary transitions, and its rise provided the ingredients for the emergence of a biosphere inhabited by complex organisms. Over the last decades, the potential for bioengineering multicellular systems has been instrumental in interrogating nature and exploring novel paths to regeneration, disease, cognition, and behaviour. Here, we provide a list of open problems that encapsulate many of the ongoing and future challenges in the field and suggest conceptual approaches that may facilitate progress.

JTD Keywords: Biology, Complexity, Differential adhesion, Evolution, Gene network model, Morphogenesis, Origin, Pattern-formation, Principles, Self-organization

The discovery that sponges (Porifera) can fully regenerate from aggregates of dissociated cells launched them as one of the earliest experimental models to study the evolution of cell adhesion and allorecognition in animals. This process depends on an extracellular glycoprotein complex called the Aggregation Factor (AF), which is composed of proteins thought to be unique to sponges. We used quantitative proteomics to identify additional AF components and interacting proteins in the classical model, Clathria prolifera, and compared them to proteins involved in cell interactions in Bilateria. Our results confirm MAFp3/p4 proteins as the primary components of the AF but implicate related proteins with calx-beta and wreath domains as additional components. Using AlphaFold, we unveiled close structural similarities of AF components to protein domains in other animals, previously masked by the mutational decay of sequence similarity. The wreath domain, believed to be unique to the AF, was predicted to contain a central beta- sandwich of the same organization as the vWFD domain (also found in extracellular, gel- forming glycoproteins in other animals). Additionally, many copurified proteins share a conserved C- terminus, containing divergent immunoglobulin (Ig) and Fn3 domains predicted to serve as an AF-interaction interface. One of these proteins, MAF- associated protein 1, resembles Ig superfamily cell adhesion molecules and we hypothesize that it may function to link the AF to the surface of cells. Our results highlight the existence of an ancient toolkit of conserved protein domains regulating cell-cell and cell-extracellular matrix protein interactions in all animals, and likely reflect a common origin of cell adhesion and allorecognition.

JTD Keywords: Adhesion, Allorecognitio, Binding, Calcium, Carbohydrate-carbohydrate interactions, Cell-cell adhesion, Evolution, Marine sponge, Microciona-prolifera, Molecule, Porifera, Protein, Proteomics, Recepto, Recognition

The diverse roles of the low-density lipoprotein receptor family (LDLR) have been associated with many processes critical to maintaining central nervous system (CNS) health and contributing to neurological diseases or cancer. In this review, we provide a comprehensive understanding of the LDLR's involvement in common brain tumors, specifically high-grade gliomas, emphasizing the receptors' critical role in the pathophysiology and progression of these tumors due to LDLR's high expression. We delve into LDLR's role in regulating cellular uptake and transport through the brain barrier. Additionally, we highlight LDLR's role in activating several signaling pathways related to tumor proliferation, migration, and invasion, engaging readers with an in-depth understanding of the molecular mechanisms at play. By synthesizing current research findings, this review underscores the significance of LDLR during tumorigenesis and explores its potential as a therapeutic target for high-grade gliomas. The collective insights presented here contribute to a deeper appreciation of LDLR's multifaceted roles and implications for physiological and pathological states, opening new avenues for tumor treatment. The role of LDLR family receptors in mediating the transport of LDL across the blood-brain barrier (BBB), facilitating processes such as survival, proliferation, and invasion in high-grade gliomas. Nanoparticles targeting LDLR can be used for drug delivery, potentially inducing cell death and reducing tumor proliferation and survival in high-grade glioma cells. image

JTD Keywords: Animals, Blood-brain-barrier, Brain neoplasms, Cancer, Delivery, Doxorubicin, Expression, Glioblastomas, Glioma, Humans, Low-density lipoprotein receptors, Low‐density lipoprotein receptors, Migratio, Nanoparticles, Neoplasm grading, Proliferation, Receptors, ldl, Signal transduction, Targeted therapie, Targeted therapies, Targeting therapy, Tumor microenvironment

The mechanics of the extracellular matrix (ECM) determine cell activity and fate through mechanoresponsive proteins including Yes-associated protein 1 (YAP). Rigidity and viscous relaxation have emerged as the main mechanical properties of the ECM steering cell behavior. However, how cells integrate coexisting ECM rigidity and viscosity cues remains poorly understood, particularly in the high-stiffness regime. Here, we have exploited engineered stiff viscoelastic protein hydrogels to show that, contrary to current models of cell-ECM interaction, substrate viscous energy dissipation attenuates mechanosensing even when cells are exposed to higher effective rigidity. This unexpected behavior is however readily captured by a pull-and-hold model of molecular clutch-based cell mechanosensing, which also recapitulates opposite cellular response at low rigidities. Consistent with predictions of the pull-and-hold model, we find that myosin inhibition can boost mechanosensing on cells cultured on dissipative matrices. Together, our work provides general mechanistic understanding on how cells respond to the viscoelastic properties of the ECM.

JTD Keywords: Adhesion, Animals, Defines, Dynamics, Elasticity, Extracellular matrix, Force, Humans, Hydrogels, Mechanotransduction, Mechanotransduction, cellular, Stability, Tali, Viscosity, Yap/taz

Electrolyte-gated organic transistors (EGOTs) leveraging organic semiconductors' electronic and ionic transport characteristics are the key enablers for many biosensing and bioelectronic applications that can selectively sense, record, and monitor different biological and biochemical processes at the nanoscale and translate them into macroscopic electrical signals. Understanding such transduction mechanisms requires multiscale characterization tools to comprehensively probe local electrical properties and link them with device behavior across various bias points. Here, an automated scanning dielectric microscopy toolbox is demonstrated that performs operando in-liquid scanning dielectric microscopy measurements on functional EGOTs and carries out extensive data analysis to unravel the evolution of local electrical properties in minute detail. This paper emphasizes critical experimental considerations permitting standardized, accurate, and reproducible data acquisition. The developed approach is validated with EGOTs based on blends of organic small molecule semiconductor and insulating polymer that work as accumulation-mode field-effect transistors. Furthermore, the degradation of local electrical characteristics at high gate voltages is probed, which is apparently driven by the destruction of local crystalline order due to undesirable electrochemical swelling of the organic semiconducting material near the source electrode edge. The developed approach paves the way for systematic probing of EGOT-based technologies for targeted optimization and fundamental understanding. This study presents automated scanning dielectric microscopy toolbox for comprehensive nanoscale electrical measurements of operating electrolyte-gated organic transistors (EGOTs). It highlights critical experimental practices for accurate data acquisition and easily reproducible analysis. The approach is validated using EGOTs based on blends of organic semiconductor and insulating polymer, revealing degradation at high gate voltages due to electrochemical swelling in minute details. image

JTD Keywords: Automation, Electrolyte-gated organic transistors, Nanoscale, Operando scanning dielectric microscopy, Transistor degradatio

Cardiac fibrosis occurs following insults to the myocardium and is characterized by the abnormal accumulation of non-compliant extracellular matrix (ECM), which compromises cardiomyocyte contractile activity and eventually leads to heart failure. This phenomenon is driven by the activation of cardiac fibroblasts (cFbs) to myofibroblasts and results in changes in ECM biochemical, structural and mechanical properties. The lack of predictive in vitro models of heart fibrosis has so far hampered the search for innovative treatments, as most of the cellular-based in vitro reductionist models do not take into account the leading role of ECM cues in driving the progression of the pathology. Here, we devised a single-step decellularization protocol to obtain and thoroughly characterize the biochemical and micro-mechanical properties of the ECM secreted by activated cFbs differentiated from human induced pluripotent stem cells (iPSCs). We activated iPSC-derived cFbs to the myofibroblast phenotype by tuning basic fibroblast growth factor (bFGF) and transforming growth factor beta 1 (TGF-beta 1) signalling and confirmed that activated cells acquired key features of myofibroblast phenotype, like SMAD2/3 nuclear shuttling, the formation of aligned alpha-smooth muscle actin (alpha- SMA)-rich stress fibres and increased focal adhesions (FAs) assembly. Next, we used Mass Spectrometry, nanoindentation, scanning electron and confocal microscopy to unveil the characteristic composition and the visco-elastic properties of the abundant, collagen-rich ECM deposited by cardiac myofibroblasts in vitro. Finally, we demonstrated that the fibrotic ECM activates mechanosensitive pathways in iPSC-derived cardiomyocytes, impacting on their shape, sarcomere assembly, phenotype, and calcium handling properties. We thus propose human bio-inspired decellularized matrices as animal-free, isogenic cardiomyocyte culture substrates recapitulating key pathophysiological changes occurring at the cellular level during cardiac fibrosis.

JTD Keywords: Adhesio, Cardiac fibrosis modelling, Decellularized extracellular matrix, Differentiation, Expression, Fibroblast activation, Fibronectin, Heart, Induced pluripotent stem cells, Ipsc-derived-cardiac fibroblasts, Ipsc-derived-cardiomyocyte, Myocardial-infarction, Neonatal cardiomyocytes, Smooth muscle actin, Substrate stiffness

Objective:To investigate the spermidine pathway capability to predict patients at risk for tumor recurrence following colorectal cancer (CRC) surgery.Background:Recurrence rates after CRC surgery remain at about 20% despite an optimal technique and adjuvant therapy when necessary. Identification of risk biomarkers of recurrence is an unmet need. The spermidine pathway is indispensable for cell proliferation and differentiation, and is suggested to accelerate tumor spread.Methods:This was a prospective cohort study of patients undergoing CRC surgery from 2015 to 2018. Plasma samples were collected before surgery and on postoperative day 4, and the spermidine pathway was assessed through mass spectrometry. Oncological outcomes were registered.Results:A total of 146 patients were included and 24 (16.4%) developed tumor recurrence. Higher levels of preoperative spermidine pathway components (spermidine, spermine, spermidine synthase enzyme, and spermine/arginine balance) were positively associated with recurrence. Surgery promoted a decrease in these pathway elements. The greater the decline was, the lower the risk of recurrence. Preoperative spermidine over the cut-off of 0.198 mu M displayed a 4.69-fold higher risk of recurrence. The spermine synthase enzyme behaved in the opposite direction.Conclusions:The spermidine pathway is associated with tumor recurrence following CRC surgery and, after confirmation in larger cohorts, could be translated as a risk biomarker of recurrence into clinical practice.

JTD Keywords: Adjuvant chemotherapy, Colon-cancer, Colorectal cancer, Energ, Metabolomics, Polyamine levels, Postoperative intraabdominal infection, Spermidine pathway, Stage, Tumor recurrenc

Biomaterial-associated infections (BAIs) pose significant challenges in modern medical technologies, being a major postoperative complication and leading cause of implant failure. These infections significantly risk patient health, resulting in prolonged hospitalization, increased morbidity and mortality rates, and elevated treatment expenses. This comprehensive review examines the mechanisms driving bacterial adhesion and biofilm formation on biomaterial surfaces, offering an in-depth analysis of current antimicrobial strategies for preventing BAIs. We explore antimicrobial-eluting biomaterials, contact-killing surfaces, and antifouling coatings, emphasizing the application of antifouling polymer brushes on medical devices. Recent advancements in multifunctional antimicrobial biomaterials, which integrate multiple mechanisms for superior protection against BAIs, are also discussed. By evaluating the advantages and limitations of these strategies, this review aims to guide the design and development of highly efficient and biocompatible antimicrobial biomaterials. We highlight potential design routes that facilitate the transition from laboratory research to clinical applications. Additionally, we provide insights into the potential of synthetic biology as a novel approach to combat antimicrobial resistance. This review aspires to inspire future research and innovation, ultimately improving patient outcomes and advancing medical device technology.

JTD Keywords: Animals, Anti-bacterial agents, Antifouling coatings, Antimicrobial peptide, Antimicrobial-eluting coatings, Antimicrobial‐eluting coatings, Bacterial adhesion, Bacterial biofilm formation, Biocompatible materials, Biofilms, Biomaterial-associated infections, Biomaterial‐associated infections, Contact killing coatings, Humans, In-vitr, Infused porous surfaces, Metal-oxide surfaces, Multifunctional antimicrobial coating, Multifunctional antimicrobial coatings, Poly(l-lysine)-g-poly(ethylene glycol) layers, Polymer brushes, Prosthesis-related infections, Silicone oil, Superhydrophobic surfaces, Urinary catheters

Hyperpolarized nuclear spins in molecules exhibit high magnetization that is unachievable by classical polarization techniques, making them widely used as sensors in physics, chemistry, and medicine. The state of a hyperpolarized material, however, is typically only studied indirectly and with partial destruction of magnetization, due to the nature of conventional detection by resonant-pickup NMR spectroscopy or imaging. Here, we establish atomic magnetometers with sub-pT sensitivity as an alternative modality to detect in real time the complex dynamics of hyperpolarized materials without disturbing or interrupting the magnetogenesis process. As an example of dynamics that are impossible to detect in real time by conventional means, we examine parahydrogen-induced H-1 and C-13 magnetization during adiabatic eigenbasis transformations at mu mu T-field avoided crossings. Continuous but nondestructive magnetometry reveals previously unseen spin dynamics, fidelity limits, and magnetization backaction effects. As a second example, we apply magnetometry to observe the chemical-exchange-driven C-13 hyperpolarization of [1-C-13]-pyruvate-the most important spin tracer for clinical metabolic imaging. The approach can be readily combined with other high-sensitivity magnetometers and is applicable to a broader range of general observation scenarios involving production, transport, and systems interaction of hyperpolarized compounds.

JTD Keywords: Adiabaticity, Dnp, Field, Hyperpolarization, Nmr, Nuclear spins, Optical magnetometry, Order, Para-hydrogen, Parahydrogen, Polarimetry, Polarization transfer

Mesenchymal stem cells (MSCs) interact with their surroundings via integrins, which link to the actin cytoskeleton and translate physical cues into biochemical signals through mechanotransduction. N-cadherins enable cell-cell communication and are also linked to the cytoskeleton. This crosstalk between integrins and cadherins modulates MSC mechanotransduction and fate. Here we show the role of this crosstalk in the mechanosensing of viscosity using supported lipid bilayers as substrates of varying viscosity. We functionalize these lipid bilayers with adhesion peptides for integrins (RGD) and N-cadherins (HAVDI), to demonstrate that integrins and cadherins compete for the actin cytoskeleton, leading to an altered MSC mechanosensing response. This response is characterised by a weaker integrin adhesion to the environment when cadherin ligation occurs. We model this competition via a modified molecular clutch model, which drives the integrin/cadherin crosstalk in response to surface viscosity, ultimately controlling MSC lineage commitment. The crosstalk between cell-cell and cell-matrix adhesions regulates stem cell fate. Here, the authors reveal a critical role for matrix viscosity in controlling this crosstalk, which they explain via a modified molecular clutch model.

JTD Keywords: Actin cytoskeleton, Adhesion, Animals, Arginyl-glycyl-aspartic acid, Cadherins, Cell adhesion, Cell communication, Fibronectin, Force transmission, Humans, Hydrogel, Integrins, Lipid bilayers, Matrix, Mechanotransduction, Mechanotransduction, cellular, Mesenchymal stem cells, Mobility, Oligopeptides, Osteogenic differentiation, Substrate stiffness, Vinculin, Viscosity

Herein, a facile approach toward converting a three-dimensional polyurethane sponge (PUS), employed in endoluminal vacuum-assisted closure (endo-VAC) therapies, in a theranostic material able to detect and to inhibit bacteria growth, has been reported. The endo-VAC PUS presented sensitivity to Gram-positive and Gramnegative bacterial species thanks to its functionalization with gold and silver antibacterial nanoparticles (NPs). PUS with chitosan-stabilized Au-NPs achieved 5.26 f 0.17 logs and 2.78 f 0.34 logs of reduction of bacteria growth, whereas the sponges functionalized with phenolated lignin Ag-NPs offered slightly inferior values (4.77 f 0.36 logs and 2.03 f 0.37 logs, respectively), against Escherichia coli and Staphylococcus aureus pathogens, respectively, after the application of photothermal ablation. The in vitro antimicrobial studies were contrasted with the in-situ monitoring of bacteria localization and inactivation with excitation lasers of 532 and 785 nm wavelengths, respectively, in the Raman equipment. The novel theranostic nano-enabled antimicrobial PU sponges offer unprecedented possibilities for the improvement of the endo-VAC treatments and extrapolation of the methodology to other plastic-based implants to combat antimicrobial resistances.

JTD Keywords: Adhesiv, Gold nanoparticles, Lignin, Molecular-mechanism, Polyurethane, Silver nanoparticles, Surface activation, Surface enhanced raman scatterin

Endothelium, the lining in this blood vessel, orchestrates three main critical functions such as protecting blood components, modulating of hemostasis by secreting various inhibitors, and directing clot digestion (fibrinolysis) by activating tissue plasminogen activator. No other surface can perform these tasks; thus, the contact of blood and blood-contacting medical devices inevitably leads to the activation of coagulation, often causing device failure, and thromboembolic complications. This perspective, first, discusses the biological mechanisms of activation of coagulation and highlights the efforts of advanced coatings to recapitulate one characteristic of endothelium, hereafter single functions of endothelium and noting necessity of the synergistic integration of its three main functions. Subsequently, it is emphasized that to overcome the challenges of blood compatibility an endothelium-mimicking system is needed, proposing a synergy of bottom-up synthetic biology, particularly synthetic cells, with passive- and bioactive surface coatings. Such integration holds promise for developing advanced biomaterials capable of recapitulating endothelial functions, thereby enhancing the hemocompatibility and performance of blood-contacting medical devices. The activation of coagulation on the surface of blood-contacting medical devices often leads to thromboembolic complications. A concept for the next generation of hemocompatbile surfaces inspired by endothelium is proposed. This concept not only contribute to the fundamental understanding of hemocompatibility but also offer practical implications for the design and development of biomedical devices with enhanced biocompatibility and functionality. image

JTD Keywords: Animals, Antifouling coatings, Antifouling polymer brushes, Biocompatible materials, Blood coagulation, Coagulation-factor-xii, Coated materials, biocompatible, Endothelium, Endothelium, vascular, Endothelium-inspired, Endothelium‐inspired, Hemocompatibility, Hemocompatible surface coatings, Hemostasis, Heparin-induced thrombocytopenia, Humans, Nitric-oxide release, Of-the-art, Peptide macrocycle inhibitor, Plasma contact system, Protein-adsorption, Self-assembled monolayers, Surface modificatio, Synthetic endotheliu, Synthetic endothelium

Porous zirconia scaffolds manufactured using polymer-infiltrated ceramic network (PICN) and additive manufacturing technologies are emerging as promising alternatives to traditional ceramic materials in dental restorations. However, incomplete osseointegration and bacterial infections still represent challenges for the long-term performance of this new composite material. To address this, the present study aims to investigate the effect of peptide biofunctionalization on the biological performance of infiltrated zirconia scaffold surfaces. The samples used in the work consisted of a 3D-printed zirconia scaffold infiltrated with a dimethacrylate copolymer. Surface biofunctionalization was achieved using a synthetic platform containing the cell-adhesive sequence RGD and the antibacteria LF1-11 peptide (RGD-LF). The attachment of the molecule was characterized through fluorescence confocal laser scanning microscopy and X-ray photoelectron spectroscopy. The biological performance of the samples was evaluated in terms of human mesenchymal stem cell adhesion and early attachment of S. aureus. The physicochemical characterization verified the successful anchoring of the biomolecule to the surface, leading to a peptide density of 288 pmol/cm2. 2 . The biological assays confirmed the potential of RGD-LF to improve cell adhesion and spreading. In this sense, the average cell area increased fourfold in the biofunctionalized surface. Regarding bacterial adhesion, it was demonstrated that RGD-LF significantly inhibited it, reducing early adhesion by half compared to the untreated surface. Overall, this study provides valuable insights into the biofunctionalization of polymer-infiltrated 3D scaffolds for the development of cell-instructive and antibacterial surfaces tailored for dental applications.

JTD Keywords: Antibacteria, Biocompatibility, Cell adhesion, Composite, Dental ceramics, Direct ink writing, Fabrication, Infiltrated zirconia scaffolds, Mechanical-properties, Peptide biofunctionalization, Picn material, Strength, Surface, Topograph, Wear behavior

The cellular prion protein (PrP (c)) plays many roles in the developing and adult brain. In addition, PrP (c) binds to several amyloids in oligomeric and prefibrillar forms and may act as a putative receptor of abnormal misfolded protein species. The role of PrP (c) in tau seeding and spreading is not known. In the present study, we have inoculated well-characterized sarkosyl-insoluble fractions of sporadic Alzheimer's disease (sAD) into the brain of adult wild-type mice (Prnp(+/+)), Prnp(0/0) (ZH3 strain) mice, and mice over-expressing the secreted form of PrP (c) lacking their GPI anchor (Tg44 strain). Phospho-tau (ptau) seeding and spreading involving neurons and oligodendrocytes were observed three and six months after inoculation. 3Rtau and 4Rtau deposits from the host tau, as revealed by inoculating Mapt(0/0) mice and by using specific anti-mouse and anti-human tau antibodies suggest modulation of exon 10 splicing of the host mouse Mapt gene elicited by exogenous sAD-tau. However, no tau seeding and spreading differences were observed among Prnp genotypes. Our results show that PrP (c) does not affect tau seeding and spreading in vivo.

JTD Keywords: Alpha-synuclein, Alzheimer disease, Alzheimer's disease, Alzheimer’s disease, Amyloid-beta oligomers, Animals, Brain, Disease models, animal, Expression, Humans, Impairmen, Mapt, Mice, Mice, transgenic, Neurons, Paired helical filaments, Pathological tau, Prion proteins, Prnp, Prnp protein, mouse, Propagation, Prp (c), Prpc, Prpc proteins, Sarcosine, Sarkosyl, Seeding, Spreadin, Spreading, Synaptic plasticity, Tau, Tau proteins, Tauopathies

This study investigates the synthesis and optimization of nanobots (NBs) loaded with pDNA using the layer-by-layer (LBL) method and explores the impact of their collective motion on the transfection efficiency. NBs consist of biocompatible and biodegradable poly(lactic-co-glycolic acid) (PLGA) nanoparticles and are powered by the urease enzyme, enabling autonomous movement and collective swarming behavior. In vitro experiments were conducted to validate the delivery efficiency of fluorescently labeled NBs, using two-dimensional (2D) and three-dimensional (3D) cell models: murine urothelial carcinoma cell line (MB49) and spheroids from human urothelial bladder cancer cells (RT4). Swarms of pDNA-loaded NBs showed enhancements of 2.2- to 2.6-fold in delivery efficiency and 6.8- to 8.1-fold in material delivered compared to inhibited particles (inhibited enzyme) and the absence of fuel in a 2D cell culture. Additionally, efficient intracellular delivery of pDNA was demonstrated in both cell models by quantifying and visualizing the expression of eGFP. Swarms of NBs exhibited a >5-fold enhancement in transfection efficiency compared to the absence of fuel in a 2D culture, even surpassing the Lipofectamine 3000 commercial transfection agent (cationic lipid-mediated transfection). Swarms also demonstrated up to a 3.2-fold enhancement in the amount of material delivered in 3D spheroids compared to the absence of fuel. The successful transfection of 2D and 3D cell cultures using swarms of LBL PLGA NBs holds great potential for nucleic acid delivery in the context of bladder treatments.

JTD Keywords: Animals, Barrier, Cell line, tumor, Dna, Drug delivery, Drug-delivery, Enzyme catalysis, Gene delivery, Gene transfer techniques, Humans, Lactic acid, Mice, Nanobots, Nanoparticles, Pdna, Plasmids, Polyglycolic acid, Polylactic acid-polyglycolic acid copolymer, Swarming, Transfectio, Transfection, Urease, Urinary bladder neoplasms

Hydrogels with cell adhesive moieties stand out as promising materials to enhance tissue healing and regeneration. Nonetheless, bacterial infections of the implants represent an unmet major concern. In the present work, we developed an alginate hydrogel modified with a multifunctional peptide containing the RGD cell adhesive motif in combination with an antibacterial peptide derived from the 1-11 region of lactoferrin (LF). The RGD-LF branched peptide was successfully anchored to the alginate backbone by carbodiimide chemistry, as demonstrated by 1H NMR and fluorescence measurements. The functionalized hydrogel presented desirable physicochemical properties (porosity, swelling and rheological behavior) to develop biomaterials for tissue engineering. The viability of mesenchymal stem cells (MSCs) on the peptide-functionalized hydrogels was excellent, with values higher than 85 % at day 1, and higher than 95 % after 14 days in culture. Moreover, the biological characterization demonstrated the ability of the hydrogels to significantly enhance ALP activity of MSCs as well as to decrease bacterial colonization of both Gram-positive and Gram-negative models. Such results prove the potential of the functionalized hydrogels as novel biomaterials for tissue engineering, simultaneously displaying cell adhesive activity and the capacity to prevent bacterial contamination, a dual bioactivity commonly not found for these types of hydrogels. In this work we report on the functionalization of an alginate hydrogel with a tailor-made multifunctional peptide containing the cell adhesive RGD motif and the LF1-11 antibacterial peptide. Such novel multifunctional biomaterial ensures the viability of human mesenchymal stem cells, enhances ALP activity and decreases bacterial infections of both Gram-positive and Gram-negative models. image

JTD Keywords: Alginate hydrogel, Alginates, Anti-bacterial agents, Antimicrobial peptid, Antimicrobial peptide, Antimicrobial peptides, Arginyl-glycyl-aspartic acid, Biocompatible materials, Biofunctionalization, Bone, Cell adhesion, Cell survival, Composite hydrogels, Cross-linking, Hlf1-11 peptide, Human lactoferrin, Humans, Hydrogels, Immobilization, Mesenchymal stem cells, Multifunctional peptide, Oligopeptides, Peptides, Physical-properties, Scaffolds, Surfac, Tissue engineering

The precise mechanisms underlying the cellular response to static electric cues remain unclear, limiting the design and development of biomaterials that utilize this parameter to enhance specific biological behaviours. To gather information on this matter we have explored the interaction of collagen type-I, the most abundant mammalian extracellular protein, with poly(vinylidene fluoride) (PVDF), an electroactive polymer with great potential for tissue engineering applications. Our results reveal significant differences in collagen affinity, conformation, and interaction strength depending on the electric charge of the PVDF surface, which subsequently affects the behaviour of mesenchymal stem cells seeded on them. These findings highlight the importance of surface charge in the establishment of the material-protein interface and ultimately in the biological response to the material. The development of new tissue engineering strategies relies heavily on the understanding of how biomaterials interact with biological tissues. Although several factors drive this process and their driving principles have been identified, the relevance and mechanism by which the surface potential influences cell behaviour is still unknown. In our study, we investigate the interaction between collagen, the most abundant component of the extracellular matrix, and poly(vinylidene fluoride) with varying surface charges. Our findings reveal substantial variations in the binding forces, structure and adhesion of collagen on the different surfaces, which collectively explain the differential cellular responses. By exposing these differences, our research fills a critical knowledge gap and paves the way for innovations in material design for advanced tissue regeneration strategies. (c) 2024 Acta Materialia Inc. Published by Elsevier Ltd. All rights are reserved, including those for text and data mining, AI training, and similar technologies.

JTD Keywords: Adhesion, Atomic-force microscope, Biomaterials, Collagen, Collagen fibril, Electroactive material, Energ, Nanofibers, Osteogenic differentiation, Polyvinylidene fluoride, Pvdf, Stimuli, Surface charge, Surface coating, Systems

Several pathogenic Escherichia coli strains cause diarrhea. Enteroaggregative E. coli (EAEC) strains are one of the diarrheagenic pathotypes. EAEC cells form a "stacked-brick" arrangement over the intestinal epithelial cells. EAEC isolates express, among other virulence determinants, the AggR transcriptional activator and the aggregative adherence fimbriae (AAF). Overexpression of the aggR gene results in increased expression of virulence factors such as the aff genes, as well as several genes involved in specific metabolic pathways such as fatty acid degradation (fad) and arginine degradation (ast). To support the hypothesis that induction of the expression of some of these pathways may play a role in EAEC virulence, in this study we used a murine infection model to evaluate the impact of the expression of these pathways on infection parameters. Mice infected with a mutant derivative of the EAEC strain 042, characterized by overexpression of the aggR gene, showed increased disease symptoms compared to those exhibited by mice infected with the wild type (wt) strain 042. Several of these symptoms were not increased when the infecting mutant, which overexpressed aggR, lacked the fad and ast pathways. Therefore, our results support the hypothesis that different metabolic pathways contribute to EAEC virulence.

JTD Keywords: Adherence, Aggr, Burde, Chain fatty-acids, Children, Enteroaggregative e. coli, Escherichia-coli, Etiology, Infection, Mice, Microbiota, Persistent diarrhea, Protein, Scfa, Sex-differences

Diabetes is a metabolic disorder caused by the body's inability to produce or use insulin. Considering the figures projected by the World Health Organization, research on insulin therapy is crucial. Hence, we present a soft biointerface based on a thiol-yne poly(ethylene glycol) (PEG) click-hydrogel as an advanced treatment option to administrate insulin. Most importantly, the device is rendered electroactive by incorporating biocompatible poly(3,4-ethylenedioxythiophene) nanoparticles (PEDOT NPs) as conductive moieties to precisely control the release of insulin over an extended period through electrochemical stimulation (ES). The device has been carefully optimized on account of: (i) the main interactions established between PEDOT- and PEG-based moieties, which have been studied by density functional theory calculations, and reveal the choice of 4-arm PEG precursors as most suitable cross-linkers; and (ii) the concentration of PEDOT NPs in the device, which has been determined considering minimal interference with the gelation process, as well as the resulting morphological, mechanical, electrochemical, and cytocompatible properties of the PEG-based click-hydrogels. Finally, the management over insulin delivery through ES is verified in vitro, with released insulin being detected by high-performance liquid chromatography. Overall, our hydrogel-based device establishes a method for controlled insulin delivery with the potential for translation to other relevant bioelectronic applications.

JTD Keywords: Bioelectronic, Chemistry, Disease, Electroactive click-hydrogel, Energ, Insulin delivery, Pedot nanoparticles, Thiol-yne nucleophilicaddition

The development of smart biomedical devices as efficient tools in early diagnosis and therapy monitoring has recently witnessed unprecedented growth, becoming an emerging field in biomedical engineering. Sponges for endoluminal vacuum therapy, which are intended for transmitting negative pressure as trigger for tissue regeneration and for draining infections in anastomotic leakages, are massively used implants with very complex geometry and high risk of infection. In this work, commercial polyurethane (PU) sponges have been converted into smart biomedical devices by incorporating an electrochemical sensor to monitor the growth of bacteria. Such innovative approach, which allows to track the tissue healing process avoiding further infection development, has been performed applying a three-step process: 1) activation of PU using low pressure oxygen plasma; 2) incorporation of conducting polymer (CP) nanoparticles (NPs) at the surface of the activated PU by chemical oxidative polymerization; and 3) formation of a homogeneous electroactive coating using the CP NPs obtained in 2), as growth nuclei in an electrochemical polymerization. The functionalized PU sponge is able to monitor the bacteria growth in the surrounding media by detecting the concentration of nicotinamide adenine dinucleotide (NADH) from respiration reactions in the cytosol (i.e. bacteria do not have mitochondria). Conversely, respiration in normal eukaryotic cells takes place in the mitochondria, whose double membrane is not permeable to NADH. The sensing performance of the CP-coated PU sponges (limit of detection: 0.06 mM; sensitivity: 1.21 mA/cm2) has been determined in the lab using NADH solutions, while a proof of concept have been conducted using Escherichia coli bacteria cultures.

JTD Keywords: Conducting polymer, Desig, Electrochemical coating, Esophageal cancer, Nadh, Pedot, Polyurethane functionalization, Selective detection, Sponge functionalizatio

Graphene and graphene oxide (GO), due to their unique chemical and physical properties, possess biochemical characteristics that can trigger intercellular signals promoting tissue regeneration. Clinical applications of thin GO-derived sheets have inspired the development of various tissue regeneration and repair approaches. In this study, we demonstrate that ultrathin sheets of plasma-functionalized and reduced GO, with the oxygen content ranging from 3.2 % to 22 % and the nitrogen content from 0 % to 8.3 %, retain their essential mechanical and molecular integrity, and exhibit robust potential for regenerating bone tissue and blood vessels across multiple cellular and animal models. Initially, we observed the growth of blood vessels and bone tissue in vitro using these functionalized GO sheets on human adipose-derived mesenchymal stem cells and umbilical vein endothelial cells. Remarkably, our study indicates a 2.5-fold increase in mineralization and two-fold increase in tubule formation even in media lacking osteogenic and angiogenic supplements. Subsequently, we observed the initiation, conduction, and formation of bone and blood vessels in a rat tibial osteotomy model, evident from a marked 4-fold increase in the volume of low radio-opacity bone tissue and a significant elevation in connectivity density, all without the use of stem cells or growth factors. Finally, we validated these findings in a mouse critical-size calvarial defect model (33 % higher healing rate) and a rat skin lesion model (up to 2.5-fold increase in the number of blood vessels, and 35 % increase in blood vessels diameter). This study elucidates the proosteogenic and pro-angiogenic properties of both pristine and plasma-treated GO ultrathin films. These properties suggest their significant potential for clinical applications, and as valuable biomaterials for investigating fundamental aspects of bone and blood vessel regeneration.

JTD Keywords: Adhesion, Angiogenesis, Biocompatibilit, Bone regeneratio, Coatings, Fibronectin, Graphene oxide, Growth, Mesenchymal stem-cells, Osteoblast, Osteogenic differentiation, Plasma treatment, Protein, Tissue regeneration

JTD Keywords: Extracellular vesicle, Photoporation, Pre-formation loadin

Cells can adapt their active contractile properties to switch between dynamical migratory states and static homeostasis. Collective tissue surface tension, generated among others by the cortical contractility of single cells, can keep cell clusters compact, while a more bipolar, anisotropic contractility is predominantly used by mesenchymal cells to pull themselves into the extracellular matrix (ECM). Here, we investigate how these two contractility modes relate to cancer cell escape into the ECM. We compare multicellular spheroids from a panel of breast cancer cell lines with primary tumor explants from breast and cervical cancer patients by measuring matrix contraction and cellular spreading into ECM mimicking collagen matrices. Our results in spheroids suggest that tumor aggressiveness is associated with elevated contractile traction and reduced active tissue surface tension, allowing cancer cell escape. We show that it is not a binary switch but rather the interplay between these two contractility modes that is essential during this process. We provide further evidence in patient-derived tumor explants that these two contractility modes impact cancer cells' ability to leave cell clusters within a primary tumor. Our results indicate that cellular contractility is an essential factor during the formation of metastases and thus may be suitable as a prognostic criterion for the assessment of tumor aggressiveness.

JTD Keywords: Breast-cancer, Disease, Emt, Forces, Hypothesis, Intercellular-adhesion, Myoepithelial cell, Stiffness, Wetting transition

Steroid myopathy is a clinically challenging condition exacerbated by prolonged corticosteroid use or adrenal tumors. In this study, we engineered a functional three-dimensional (3D) in vitro skeletal muscle model to investigate steroid myopathy. By subjecting our bioengineered muscle tissues to dexamethasone treatment, we reproduced the molecular and functional aspects of this disease. Dexamethasone caused a substantial reduction in muscle force, myotube diameter and induced fatigue. We observed nuclear translocation of the glucocorticoid receptor (GCR) and activation of the ubiquitin-proteasome system within our model, suggesting their coordinated role in muscle atrophy. We then examined the therapeutic potential of taurine in our 3D model for steroid myopathy. Our findings revealed an upregulation of phosphorylated AKT by taurine, effectively countering the hyperactivation of the ubiquitin- proteasomal pathway. Importantly, we demonstrate that discontinuing corticosteroid treatment was insufficient to restore muscle mass and function. Taurine treatment, when administered concurrently with corticosteroids, notably enhanced contractile strength and protein turnover by upregulating the AKT-mTOR axis. Our model not only identifies a promising therapeutic target, but also suggests combinatorial treatment that may benefit individuals undergoing corticosteroid treatment or those diagnosed with adrenal tumors.

JTD Keywords: 3d bioengineered skeletal muscle tissues, Adrenal cortex hormones, Atroph, Colocalization, Corticosteroids, Dexamethasone, Glucocorticoid-receptor, Humans, Mechanisms, Models, biological, Mtor protein, human, Muscle contraction, Muscle fibers, skeletal, Muscle strength, Muscle, skeletal, Muscular diseases, Organ size, Phosphorylation, Proteasome endopeptidase complex, Proto-oncogene proteins c-akt, Receptors, glucocorticoid, Signal transduction, Skeletal-muscle, Steroid myopathy, Steroids, Supplementation, Taurin, Taurine, Tor serine-threonine kinases, Ubiquitin

Colorectal cancer (CRC) tumors are composed of heterogeneous and plastic cell populations, including a pool of cancer stem cells that express LGR5. Whether these distinct cell populations display different mechanical properties, and how these properties might contribute to metastasis is poorly understood. Using CRC patient derived organoids (PDOs), we find that compared to LGR5- cells, LGR5+ cancer stem cells are stiffer, adhere better to the extracellular matrix (ECM), move slower both as single cells and clusters, display higher nuclear YAP, show a higher survival rate in response to mechanical confinement, and form larger transendothelial gaps. These differences are largely explained by the downregulation of the membrane to cortex attachment proteins Ezrin/Radixin/Moesin (ERMs) in the LGR5+ cells. By analyzing single cell RNA-sequencing (scRNA-seq) expression patterns from a patient cohort, we show that this downregulation is a robust signature of colorectal tumors. Our results show that LGR5- cells display a mechanically dynamic phenotype suitable for dissemination from the primary tumor whereas LGR5+ cells display a mechanically stable and resilient phenotype suitable for extravasation and metastatic growth. The mechanical properties of heterogeneous cell populations in colorectal tumors and the relevance to cancer metastasis remain not fully understood. Here, the authors suggest that the variations in malignant phenotypes between LGR5-positive cancer stem cells and LGR5-negative cells could be due to their distinct mechanical phenotypes observed in vitro, determined by the membrane to cortex attachment proteins Ezrin/Radixin/Moesin.

JTD Keywords: , , Adhesion, Deformability, E-cadherin, Erm proteins, Expression, Metastasis, Organization, Plasticit, Stem-cells, Tumor-cells

Objective: Remineralising composites with antibacterial properties may seal the cavity and prevent secondary caries. This study aimed at developing experimental flowable composites containing different concentrations of fluoride-doped calcium phosphate fillers and evaluating their remineralising and antibacterial properties. Methods: Experimental resin-based composites containing different concentrations (0-20 %) of fluoride-doped calcium phosphate fillers (VS10/VS20) were formulated. The release of calcium (Ca), phosphate (PO) and fluoride (F) ions was assessed for 30 days. Remineralisation properties were evaluated through ATR-FTIR and SEM/EDX after storage in simulated body fluid (SBF). The metabolic activity and viability of Streptococcus gordonii was also evaluated through ATP, CFU and live/dead confocal microscopy. The evaluation of specific monomer elution from the experimental composites was conducted using high-performance liquid chromatography (HPLC). Results: The composites containing VS10 showed the highest release of Ca, those containing VS20 released more F over time (p < 0.05), while there was no significant difference in terms of PO ions release between the groups (p > 0.05). A quick 7-day mineral precipitation was observed in the tested composites containing VS10 or VS20 at 10 %; these materials also showed the greatest antibacterial activity (p < 0.05). Moreover, the tested composites containing VS10 presented the lowest elution of monomers (p < 0.05). Conclusions: Innovative composites were developed with low monomers elution, evident antibacterial activity against S. gordonii and important remineralisation properties due to specific ions release.

JTD Keywords: Adhesion, Antibacterial, Apatite, Bacterial, Calcium phosphate, Caries, Demineralization, Dentistry, Elution, Enamel, Ion -release, Ion-release, Monomers, Remineralisation, Resin composite, Tissue

Bladder cancer treatment via intravesical drug administration achieves reasonable survival rates but suffers from low therapeutic efficacy. To address the latter, self-propelled nanoparticles or nanobots have been proposed, taking advantage of their enhanced diffusion and mixing capabilities in urine when compared with conventional drugs or passive nanoparticles. However, the translational capabilities of nanobots in treating bladder cancer are underexplored. Here, we tested radiolabelled mesoporous silica-based urease-powered nanobots in an orthotopic mouse model of bladder cancer. In vivo and ex vivo results demonstrated enhanced nanobot accumulation at the tumour site, with an eightfold increase revealed by positron emission tomography in vivo. Label-free optical contrast based on polarization-dependent scattered light-sheet microscopy of cleared bladders confirmed tumour penetration by nanobots ex vivo. Treating tumour-bearing mice with intravesically administered radio-iodinated nanobots for radionuclide therapy resulted in a tumour size reduction of about 90%, positioning nanobots as efficient delivery nanosystems for bladder cancer therapy.© 2024. The Author(s).

JTD Keywords: cell, drug-delivery, nanomotors, tissue, Bladder cancers, Cancer therapy, Diseases, Drug administration, Drug delivery, Enhanced diffusion, Enhanced mixing, Ex-vivo, In-vivo, Mammals, Nanobots, Nanoparticles, Nanosystems, Oncology, Positron emission tomography, Radioisotopes, Silica, Survival rate, Therapeutic efficacy, Tumor penetration, Tumors

Organ-on-chip (OOC) technology has recently emerged as a powerful tool to mimic physiological or pathophysiological conditions through cell culture in microfluidic devices. One of its main goals is bypassing animal testing and encouraging more personalized medicine. The recent incorporation of hydrogels as 3D scaffolds into microfluidic devices has changed biomedical research since they provide a biomimetic extracellular matrix to recreate tissue architectures. However, this technology presents some drawbacks such as the necessity for physical structures as pillars to confine these hydrogels, as well as the difficulty in reaching different shapes and patterns to create convoluted gradients or more realistic biological structures. In addition, pillars can also interfere with the fluid flow, altering the local shear forces and, therefore, modifying the mechanical environment in the OOC model. In this work, we present a methodology based on a plasma surface treatment that allows building cell culture chambers with abutment-free patterns capable of producing precise shear stress distributions. Therefore, pillarless devices with arbitrary geometries are needed to obtain more versatile, reliable, and biomimetic experimental models. Through computational simulation studies, these shear stress changes are demonstrated in different designed and fabricated geometries. To prove the versatility of this new technique, a blood-brain barrier model has been recreated, achieving an uninterrupted endothelial barrier that emulates part of the neurovascular network of the brain. Finally, we developed a new technology that could avoid the limitations mentioned above, allowing the development of biomimetic OOC models with complex and adaptable geometries, with cell-to-cell contact if required, and where fluid flow and shear stress conditions could be controlled. A novel methodology utilizing plasma surface treatment enables the construction of cell culture chambers featuring abutment-free patterns, facilitating the precise distribution of shear stress.

JTD Keywords: Cells, Gradients, Systems

Over the past decades, the development of nanoparticles (NPs) to increase the efficiency of clinical treatments has been subject of intense research. Yet, most NPs have been reported to possess low efficacy as their actuation is hindered by biological barriers. For instance, synovial fluid (SF) present in the joints is mainly composed of hyaluronic acid (HA). These viscous media pose a challenge for many applications in nanomedicine, as passive NPs tend to become trapped in complex networks, which reduces their ability to reach the target location. This problem can be addressed by using active NPs (nanomotors, NMs) that are self-propelled by enzymatic reactions, although the development of enzyme-powered NMs, capable of navigating these viscous environments, remains a considerable challenge. Here, the synergistic effects of two NMs troops, namely hyaluronidase NMs (HyaNMs, Troop 1) and urease NMs (UrNMs, Troop 2) are demonstrated. Troop 1 interacts with the SF by reducing its viscosity, thus allowing Troop 2 to swim more easily through the SF. Through their collective motion, Troop 2 increases the diffusion of macromolecules. These results pave the way for more widespread use of enzyme-powered NMs, e.g., for treating joint injuries and improving therapeutic effectiveness compared with traditional methods. The conceptual idea of the novel approach using hyaluronidase NMs (HyaNMs) to interact with and reduce the viscosity of the synovial fluid (SF) and urease NMs (UrNMs) for a more efficient transport of therapeutic agents in joints.image

JTD Keywords: Biological barrier, Clinical research, Clinical treatments, Collective motion, Collective motion,nanomotors,nanorobots,swarming,viscous medi, Collective motions, Complex networks, Enzymatic reaction, Enzymes, Hyaluronic acid, Hyaluronic-acid,ph,viscoelasticity,adsorption,barriers,behavior,ureas, Macromolecules, Medical nanotechnology, Nano robots, Nanomotors, Nanorobots, Swarming, Synovial fluid, Target location, Viscous media, Viscous medium

The fibrotic tumor microenvironment is a pivotal therapeutic target. Nintedanib, a clinically approved multikinase antifibrotic inhibitor, is effective against lung adenocarcinoma (ADC) but not squamous cell carcinoma (SCC). Previous studies have implicated the secretome of tumor-associated fibroblasts (TAFs) in the selective effects of nintedanib in ADC, but the driving factor(s) remained unidentified. Here we examined the role of tissue inhibitor of metalloproteinase-1 (TIMP-1), a tumor-promoting cytokine overproduced in ADC-TAFs. To this aim, we combined genetic approaches with in vitro and in vivo preclinical models based on patient-derived TAFs. Nintedanib reduced TIMP-1 production more efficiently in ADC-TAFs than SCC-TAFs through a SMAD3-dependent mechanism. Cell culture experiments indicated that silencing TIMP1 in ADC-TAFs abolished the therapeutic effects of nintedanib on cancer cell growth and invasion, which were otherwise enhanced by the TAF secretome. Consistently, co-injecting ADC cells with TIMP1-knockdown ADC-TAFs into immunocompromised mice elicited a less effective reduction of tumor growth and invasion under nintedanib treatment compared to tumors bearing unmodified fibroblasts. Our results unveil a key mechanism underlying the selective mode of action of nintedanib in ADC based on the excessive production of TIMP-1 in ADC-TAFs. We further pinpoint reduced SMAD3 expression and consequent limited TIMP-1 production in SCC-TAFs as key for the resistance of SCC to nintedanib. These observations strongly support the emerging role of TIMP-1 as a critical regulator of therapy response in solid tumors.

JTD Keywords: Cancer-associated fibroblast,fibrosis,nintedanib,non-small-cell lung cancer,smad3,therapy resistance,timp-, Cell carcinoma,breast-cancer,expression,progression,inhibitor,blockade,efficac

Applications of sodium alginate (Alg) and polyacrylic acid (PAA) hydrogels in biomedicine are well-known. These are predefined by the strength and weakness of their properties, which in turn depend on the chemical structure and the architecture of their crosslinks. In this work, Alg biopolymer has been grafted to synthetic PAA that has been chemically crosslinked using N,N '-methylene-bisacrylamide (MBA) to produce a pH responsive hydrogel with adhesive property. The double crosslinking network, which combines MBA-mediated covalent crosslinks and ionic crosslinks in Alg domains, results in an elastic modulus that resembles that of highly anisotropic and viscoelastic human skin. After addressing the influence of the dual network onto the Alg-g-PAA hydrogel properties, a prospection of its potential as an adhesive has been made considering different surfaces (rubber, paper steel, porcine skin, etc). The bonding energy onto porcine skin, 32.6 +/- 4.6 J/m2, revealed that the Alg-g-PAA hydrogel can be proposed in the biomedical field as tissue adhesive for wound healing applications. Finally, the hydrogel has been semi-interpenetrated with poly(hydroxymethyl-3,4-ethylenedioxythiophene) (PEDOT-MeOH) chains through a chemical oxidative polymerization process. The resulting hydrogel, Alg-g- PAA/PEDOT-MeOH, which is even more porous than Alg-g-PAA, in addition to being electro-responsive, maintains adhesive properties.

JTD Keywords: Adhesion properties, Adhesion properties,biomedical applications,bonding energy,dual network,conducting hydrogel, Adhesive properties, Adhesives, Biomedical applications, Biopolymers, Bonding energies, Bonding energy, Chemical bonds, Conducting hydrogels, Crosslinking, Dual network, Hydrogels, Medical applications, Methylenebisacrylamide, Poly(acrylic acid), Porcine skin, Property, Rational design,film, Sodium alginate

Background: Early detection of postoperative complications after colorectal cancer (CRC) surgery is associated with improved outcomes. The aim was to investigate early metabolomics signatures capable to detect patients at risk for severe postoperative complications after CRC surgery. Materials and methods: Prospective cohort study of patients undergoing CRC surgery from 2015 to 2018. Plasma samples were collected before and after surgery, and analyzed by mass spectrometry obtaining 188 metabolites and 21 ratios. Postoperative complications were registered with Clavien-Dindo Classification and Comprehensive Complication Index. Results: One hundred forty-six patients were included. Surgery substantially modified metabolome and metabolic changes after surgery were quantitatively associated with the severity of postoperative complications. The strongest positive relationship with both Clavien-Dindo and Comprehensive Complication Index (beta=4.09 and 63.05, P<0.001) corresponded to kynurenine/tryptophan, against an inverse relationship with lysophosphatidylcholines (LPCs) and phosphatidylcholines (PCs). Patients with LPC18:2/PCa36:2 below the cut-off 0.084 mu M/mu M resulted in a sevenfold higher risk of major complications (OR=7.38, 95% CI: 2.82-21.25, P<0.001), while kynurenine/tryptophan above 0.067 mu M/mu M a ninefold (OR=9.35, 95% CI: 3.03-32.66, P<0.001). Hexadecanoylcarnitine below 0.093 mu M displayed a 12-fold higher risk of anastomotic leakage-related complications (OR=11.99, 95% CI: 2.62-80.79, P=0.004). Conclusion: Surgery-induced phospholipids and amino acid dysregulation is associated with the severity of postoperative complications after CRC surgery, including anastomotic leakage-related outcomes. The authors provide quantitative insight on metabolic markers, measuring vulnerability to postoperative morbidity that might help guide early decision-making and improve surgical outcomes.

JTD Keywords: Acids, Anastomotic leakage, Bypass, Clinical-practice guidelines, Colon, Colorectal cancer, Metabolomics, Postoperative complications, Predict, Sepsis, Trauma, Tryptophan degradation

Malaria, caused by different species of protists of the genus Plasmodium, remains among the most common causes of death due to parasitic diseases worldwide, mainly for children aged under 5. One of the main obstacles to malaria eradication is the speed with which the pathogen evolves resistance to the drug schemes developed against it. For this reason, it remains urgent to find innovative therapeutic strategies offering sufficient specificity against the parasite to minimize resistance evolution and drug side effects. In this context, nanotechnology-based approaches are now being explored for their use as antimalarial drug delivery platforms due to the wide range of advantages and tuneable properties that they offer. However, major challenges remain to be addressed to provide a cost-efficient and targeted therapeutic strategy contributing to malaria eradication. The present work contains a systematic review of nanotechnology-based antimalarial drug delivery systems generated during the last 10 years. This article is categorized under: Therapeutic Approaches and Drug Discovery > Nanomedicine for Infectious Disease

JTD Keywords: Adjuvant system, Antimalarial activities, Antimalarial agent, Antimalarial drug, Antimalarial drugs, Antimalarials, Artemisinin resistance, Causes of death, Child, Controlled drug delivery, Diseases, Drug delivery system, Drug delivery systems, Drug interactions, Drug side-effects, Drug-delivery, Experimental modelling, Heparan-sulfate, Human, Humans, In-vitro, Malaria, Malaria vaccine, Mannosylated liposomes, Medical nanotechnology, Models, theoretical, Nanocarriers, Nanomedicine, Nanotechnology, Parasite-, Parasitics, Plasmodium, Plasmodium-falciparum malaria, Red-blood-cells, Targeted delivery, Targeted drug delivery, Theoretical model, Therapeutic strategy

From complex -mixture analysis to in vivo molecular imaging, applications of liquid -state nuclear spin hyperpolarization have expanded widely over recent years. In most cases, hyperpolarized solutions are generated and transported from the polarization instrument to the measurement device. The sample hyperpolarization usually survives this transport, since the changes in magnetic fields that are external to the sample are typically adiabatic (slow) with respect to the internal nuclear spin dynamics. The passage of polarized samples through weakly magnetic components such as stainless steel syringe needles and ferrules is not always adiabatic, can lead to near -complete destruction of the magnetization. To avoid this effect becoming "folklore"in field of hyperpolarized NMR, we present a systematic investigation to highlight the problem and investigate possible solutions. Experiments were carried out on: (i) dissolution-DNP-polarized [1-13C]pyruvate with detection at 1.4 T, and (ii) 1.5 -T -polarized H2O with NMR detection at 2.5 mu T. We show that the degree adiabaticity of solutions passing through metal parts is intrinsically unpredictable, likely depending on factors such as solution flow rate, degree of remanent ferromagnetism in the metal, and nuclear spin However, the magnetization destruction effects can be suppressed by application of an external field order of 0.1-10 mT.

JTD Keywords: Benchtop nmr, Hyperpolarization, Low-field mri, Non-adiabatic, Para-hydrogen, Spin relaxation

Microbial biofilms are complex three-dimensional structures where sessile microbes are embedded in a polymeric extracellular matrix. Their resistance toward the host immune system as well as to a diverse range of antimicrobial treatments poses a serious health and development threat, being in the top 10 global public health threats declared by the World Health Organization. In an effort to combat biofilm-related microbial infections, several strategies have been developed to independently eliminate biofilms or to complement conventional antibiotic therapies. However, their limitations leave room for other treatment alternatives, where the application of nanotechnology to biofilm eradication has gained significant relevance in recent years. Their small size, penetration efficiency, and the design flexibility that they present makes them a promising alternative for biofilm infection treatment, although they also present set-backs. This review aims to describe the main possibilities and limitations of nanomedicine against biofilms, while covering the main aspects of biofilm formation and study, and the current therapies for biofilm treatment. This article is categorized under: Therapeutic Approaches and Drug Discovery > Nanomedicine for Infectious Disease Toxicology and Regulatory Issues in Nanomedicine > Toxicology of Nanomaterials Toxicology and Regulatory Issues in Nanomedicine > Regulatory and Policy Issues in Nanomedicine.

JTD Keywords: Anti-bacterial agents, Anti-infective agents, Antiinfective agent, Antimicrobial, Antimicrobials, Antimicrobials,bacteria,biofilm,infectious diseases,microorganism, Bacteria, Biofilm, Biofilm infections, Biofilms, Complex three dimensional structures, Diseases, Diverse range, Drug-delivery systems,in-vitro,cellular toxicity,nanoparticles,penetration,model,biocompatibility,perspectives,hyperthermia,diagnosi, Extracellular matrices, Global public health, Health risks, Infectious disease, Infectious diseases, Medical nanotechnology, Microbial biofilm, Microorganisms, Nanomedicine, Polymer, Polymers, Regulatory issues, Roadmap

Tissue engineering holds great promise for biomedical research and healthcare, offering alternatives to animal models and enabling tissue regeneration and organ transplantation. Three-dimensional (3D) bioprinting stands out for its design flexibility and reproducibility. Here, we present an integrated fluorescent light sheet bioprinting and imaging system that combines high printing speed (0.66 mm3 /s) and resolution (9 μm) with light sheet-based imaging. This approach employs direct laser patterning and a static light sheet for confined voxel crosslinking in photocrosslinkable materials. The developed bioprinter enables real-time monitoring of hydrogel crosslinking using fluorescent recovery after photobleaching (FRAP) and brightfield imaging as well as in situ light sheet imaging of cells. Human fibroblasts encapsulated in a thiol-ene click chemistry-based hydrogel exhibited high viability (83% ± 4.34%) and functionality. Furthermore, full-thickness skin constructs displayed characteristics of both epidermal and dermal layers and remained viable for 41 days. The integrated approach demonstrates the capabilities of light sheet bioprinting, offering high speed, resolution, and real-time characterization. Future enhancements involving solid-state laser scanning devices such as acousto-optic deflectors and modulators will further enhance resolution and speed, opening new opportunities in light-based bioprinting and advancing tissue engineering. This article is protected by copyright. All rights reserved.This article is protected by copyright. All rights reserved.

JTD Keywords: cadherin, collagen, culture, differentiation, fluorescence microscopy, full-thickness skin model, hydrogels, light sheet bioprinter, light sheet fluorescence microscopy, proliferation, survival, tissue engineering, Animal, Animals, Biofabrication, Bioprinting, Cell culture, Crosslinking, Fluorescence, Fluorescence microscopy, Full-thickness skin model, Hair follicle, Human, Humans, Hydrogel, Hydrogels, Image resolution, Laser patterning, Light sheet, Light sheet bioprinter, Light sheet fluorescence microscopy, Molecular biology, Photobleaching, Printing, three-dimensional, Procedures, Reproducibility, Reproducibility of results, Skin model, Three dimensional printing, Tissue, Tissue engineering, Tissue regeneration, Tissue scaffolds, Tissues engineerings

Exposure of experimental rodents to controlled cycles of light, food, and temperature is important when investigating alterations in circadian cycles that profoundly influence health and disease. However, applying such stimuli simultaneously is difficult in practice. We aimed to design, build, test, and open-source describe a simple device that subjects a conventional mouse cage to independent cycles of physiologically relevant environmental variables. The device is based on a box enclosing the rodent cage to modify the light, feeding, and temperature environments. The device provides temperature-controlled air conditioning (heating or cooling) by a Peltier module and includes programmable feeding and illumination. All functions are set by a user-friendly front panel for independent cycle programming. Bench testing with a model simulating the CO2 production of mice in the cage showed: a) suitable air renewal (by measuring actual ambient CO2), b) controlled realistic illumination at the mouse enclosure (measured by a photometer), c) stable temperature control, and d) correct cycling of light, feeding, and temperature. The cost of all the supplies (retail purchased by e-commerce) was <300 US$. Detailed technical information is open-source provided, allowing for any user to reliably reproduce or modify the device. This approach can considerably facilitate circadian research since using one of the described low-cost devices for any mouse group with a given light-food-temperature paradigm allows for all the experiments to be performed simultaneously, thereby requiring no changes in the light/temperature of a general-use laboratory. 1 Introduction

JTD Keywords: Animal experiment, Animal model, Animal research, Article, Circadian alteration, Circadian rhythm, Commercial phenomena, Controlled study, Cycling, Energy consumption, Energy-expenditure, Experimental model, Feeding, Food, Food availability, Illumination, Intermittent fasting, Light, Light cycle, Light dark cycle, Mouse, Nonhuman, Open source technology, Open-source hardware, Performance, Photography, Research, Rhythms, Rodent, Temperature, Temperature cycle

Polarized hydroxyapatite (p-HAp) has been used as a catalyst for the synthesis of urea coupling N-2, CO2 and water under mild reaction conditions when compared to classical nitrogen fixation reactions, such as the Haber-Bosch process. The reaction of 3 bar of N-2 and 3 bar of CO2 under UV illumination at 120 degrees C (for 48 h) results in a urea yield of 1.5 +/- 0.1 mmol per gram of catalyst (g(c)) with a selectivity close to 80%, whereas the reaction is not successful without UV irradiation. However, the addition of small amounts of NO (314 ppm) produces 15.2 +/- 0.6 and 4.6 +/- 0.4 mmol g(c)(-1) with and without UV illumination, respectively, with the selectivity in both cases being close to 100%. As nitrogen fixation without UV irradiation using p-HAp as a catalyst is a challenge, studies with NO have been conducted varying the reaction conditions (time, pressure and temperature). The results suggest a mechanism based on the production of NH4+ through the oxidation of N-2.

JTD Keywords: Carbon dioxide, Carbon,dinitrogen,reduction,nitrogen,ammonia,dioxid, Catalyst selectivity, Condition, Haber-bosch process, Hydroxyapatite, Irradiation, Metabolism, Mild reaction conditions, Nitrogen fixation, Pressure and temperature, Reaction conditions, Time pressures, Time-temperature, Urea, Uv illuminations, Without uv irradiations, ]+ catalyst

Factor XII (FXII) is a zymogen present in blood that tends to adsorb onto the surfaces of blood-contacting medical devices. Once adsorbed, it becomes activated, initiating a cascade of enzymatic reactions that lead to surface-induced coagulation. This process is characterized by multiple redundancies, making it extremely challenging to prevent clot formation and preserve the properties of the surface. In this study, a novel modulatory coating system based on C1-esterase inhibitor (C1INH) functionalized polymer brushes, which effectively regulates the activation of FXII is proposed. Using surface plasmon resonance it is demonstrated that this coating system effectively repels blood plasma proteins, including FXII, while exhibiting high activity against activated FXII and plasma kallikrein under physiological conditions. This unique property enables the modulation of FXII activation without interfering with the overall hemostasis process. Furthermore, through dynamic Chandler loop studies, it is shown that this coating significantly improves the hemocompatibility of polymeric surfaces commonly used in medical devices. By addressing the root cause of contact activation, the synergistic interplay between the antifouling polymer brushes and the modulatory C1INH is expected to lay the foundation to enhance the hemocompatibility of medical device surfaces.© 2023 The Authors. Macromolecular Bioscience published by Wiley-VCH GmbH.

JTD Keywords: adsorption, binding, c1-esterase-inhibitor, coatings, contact activation, factor-xii, fxii activation, hemocompatibility, hemocompatible surface modification, heparin, polymer brushes, system, thrombosis, Adsorption, Anticoagulation, Antifouling agent, Article, Beta-fxiia, Biocompatibility, Blood, Blood clotting, Blood clotting factor 12, Blood clotting factor 12a, Blood clotting factor 12a inhibitor, Blood coagulation, C1-esterase-inhibitor, Cell activation, Chemical activation, Coagulation, Coating (procedure), Complement component c1s inhibitor, Complement system, Controlled study, Dendrimers, Enzyme immobilization, Enzymes, Erythrocyte, Esters, Factor xii, Factor xii activation, Factor xiia, Fibrin deposition, Functional polymers, Fxii activation, Haemocompatibility, Hemocompatibility, Hemocompatible surface modification, Hemostasis, Heparin, Human, Hydrogel, Medical devices, Metabolism, Plasma kallikrein, Plasma protein, Plastic coatings, Platelet count, Polymer, Polymer brushes, Polymerization, Polymers, Property, Root cause, Surface plasmon resonance, Surface property, Surface reactions, Surface-modification, Thrombocyte adhesion, Β-fxiia

A new generation of fully bioresorbable metallic Zn-based alloys could be used for stenting applications; however, the initial surface degradation delays stent re-endothelialization. Thus, this work proposes a dual strategy to control the corrosion and accelerate the endothelialization of ZnMg and ZnAg biodegradable alloys. First, a stable polycaprolactone (PCL) coating is obtained and followed by its functionalization with either linear RGD (Arg-Gly-Asp) or REDV (Arg-Glu-Asp-Val) peptides or a dual peptide-based platform combining both sequences (RGD-REDV). Scratching tests showed neither delamination nor detachment of the polymeric coating. Potentiodynamic polarization (PDP) and electrochemical impedance spectroscopy (EIS) measurements confirmed the corrosion resistance after PCL coating by revealing lower current density and higher absolute impedance values. X-ray photoelectron spectroscopy (XPS) and fluorescent microscopy confirmed the correct peptide immobilization onto PCL coated Zn alloys. The functionalized samples exhibited enhanced human umbilical vein endothelial cells (HUVEC) adhesion. The higher number of adhered cells to the functionalized surfaces with the RGD-REDV platform demonstrates the synergistic effect of combining both RGD and REDV sequences. Higher corrosion resistance together with enhanced endothelialization indicates that the dual functionalization of Zn alloys with PCL and peptide-based RGD-REDV platform holds great potential to overcome the clinical limitations of current biodegradable metal stents.

JTD Keywords: Binary alloys, Biodegradable metals, Bioresorbable, Cardiovascular applications, Cell adhesive peptides, Corrosion, Corrosion resistance, Corrosion resistant alloys, Corrosion resistant coatings, Degradation, Dual peptide-based platform, Electrochemical corrosion, Electrochemical impedance spectroscopy, Endothelial cells, Endothelialization, Functionalization, Functionalizations, In-vitro, Magnesium alloys, Metallics, Mg alloy, Peptides, Polycaprolactone coating, Polymer-coatings, Rgd-functionalization, Silver alloys, Stents, Surface, X ray photoelectron spectroscopy, Zinc, Zinc alloys, Zn alloys, Zn-based alloys

Interactions between living cells and nanoparticles are extensively studied to enhance the delivery of therapeutics. Nanoparticles size, shape, stiffness, and surface charge are regarded as the main features able to control the fate of cell-nanoparticle interactions. However, the clinical translation of nanotherapies has so far been limited, and there is a need to better understand the biology of cell-nanoparticle interactions. This study investigates the role of cellular mechanosensitive components in cell-nanoparticle interactions. It is demonstrated that the genetic and pharmacologic inhibition of yes-associated protein (YAP), a key component of cancer cell mechanosensing apparatus and Hippo pathway effector, improves nanoparticle internalization in triple-negative breast cancer cells regardless of nanoparticle properties or substrate characteristics. This process occurs through YAP-dependent regulation of endocytic pathways, cell mechanics, and membrane organization. Hence, the study proposes targeting YAP may sensitize triple-negative breast cancer cells to chemotherapy and increase the selectivity of nanotherapy.© 2023 The Authors. Advanced Science published by Wiley-VCH GmbH.

JTD Keywords: cancer treatment, cells, differentiation, hippo pathway, mechanics, mechanobiology, mechanotransduction, nanoparticles, progression, protein, resistance, yap-signaling, yap/taz, Adaptor proteins, signal transducing, Bio-nano interaction, Bio-nano interactions, Breast cancer cells, Cancer cells, Cancer treatment, Cells, Cellular therapeutics, Cellular uptake, Chemotherapy, Cytology, Diseases, Extracellular-matrix, Human, Humans, Mechano-biology, Mechanobiology, Metabolism, Nanoparticle, Nanoparticle interaction, Nanoparticles, Physiology, Protein serine threonine kinase, Protein serine-threonine kinases, Protein signaling, Signal transducing adaptor protein, Signal transduction, Therapeutic effects, Triple negative breast cancer, Triple negative breast neoplasms, Triple-negative breast cancers, Yap-signaling, Yap-signaling proteins, Yes-associated protein-signaling

Circadian rhythms are archetypal examples of nonlinear oscillations. While these oscillations are usually attributed to circuits of biochemical interactions among clock genes and proteins, recent experimental studies reveal that they are also affected by the cell's mechanical environment. Here, we extend a standard biochemical model of circadian rhythmicity to include mechanical effects in a parametric manner. Using experimental observations to constrain the model, we suggest specific ways in which the mechanical signal might affect the clock. Additionally, a bifurcation analysis of the system predicts that these mechanical signals need to be within an optimal range for circadian oscillations to occur.

JTD Keywords: Circadian rhythm, Clocks, Coherence, Synchronization

Although we have learned much about how the brain fuels its functions over the last decades, there remains much still to discover in an organ that is so complex. This article lays out major gaps in our knowledge of interrelationships between brain metabolism and brain function, including biochemical, cellular, and subcellular aspects of functional metabolism and its imaging in adult brain, as well as during development, aging, and disease. The focus is on unknowns in metabolism of major brain substrates and associated transporters, the roles of insulin and of lipid droplets, the emerging role of metabolism in microglia, mysteries about the major brain cofactor and signaling molecule NAD+, as well as unsolved problems underlying brain metabolism in pathologies such as traumatic brain injury, epilepsy, and metabolic downregulation during hibernation. It describes our current level of understanding of these facets of brain energy metabolism as well as a roadmap for future research. This article details current knowledge and major unknowns in brain energy metabolism and lays out a roadmap for future research.image

JTD Keywords: Acetate, Acetyl-coa, Aerobic glycolysis, Atp-citrate lyase, Extracellular glutamate concentration, Fatty-acid transport, Glucose-metabolism, Glut4, In-vivo, Insulin, Lipid droplet accumulation, Nicotinamide adenine-dinucleotide, Noradrenaline, Obese zucker rats, Rat cerebral-cortex

Very recently, the controlled release of dopamine (DA), a neurotransmitter whose deficiency is associated with Parkinson's disease, has been postulated as a good alternative to the oral administration of levodopa (L-Dopa), a dopamine precursor, to combat the effects of said disease. However, this is still a very little explored field and there are very few carriers that are capable of releasing DA, a small and water-soluble molecule, in an efficient and controlled manner. In this work, we report a carrier based on a conductive hydrogel capable of loading DA and releasing it progressively and efficiently (100 % release) in a period of five days by applying small electrical stimuli (-0.4 V) daily for a short time (1 min). The hydrogel (CMC/PEDOT), which is electrically active, has been prepared from sodium carboxymethylcellulose and poly(3,4-ethylenedioxythiophene) microparticles, using citric acid as a cross-linking agent. Furthermore, the results have shown that when relatively hydrophobic small molecules, such as chloramphenicol, are loaded, the electrostimulated release is significantly less efficient, demonstrating the usefulness of CMC/PEDOT as a carrier for neurotransmitters.

JTD Keywords: Amines, Carboxymethyl cellulose, Carboxymethylcellulose, Conducting hydrogels, Conducting polymers, Controlled release, Crosslinking, Dopamine, Drug-delivery system, Electrostimulation, Hydrogels, Joining, Levodopa, Loading, Molecules, Neurophysiology, Neurotransmitter release, Neurotransmitters release, Oral administration, Parkinson's disease, Parkinsons-disease, Poly(3,4-ethylenedioxythiophene), Release, Sodium, Transport, Water-soluble molecule

Human pluripotent stem cell -derived kidney organoids offer unprecedented opportunities for studying polycystic kidney disease (PKD), which still has no effective cure. Here, we developed both in vitro and in vivo organoid models of PKD that manifested tubular injury and aberrant upregulation of renin-angiotensin aldosterone system. Single -cell analysis revealed that a myriad of metabolic changes occurred during cystogenesis, including defective autophagy. Experimental activation of autophagy via ATG5 overexpression or primary cilia ablation significantly inhibited cystogenesis in PKD kidney organoids. Employing the organoid xenograft model of PKD, which spontaneously developed tubular cysts, we demonstrate that minoxidil, a potent autophagy activator and an FDA -approved drug, effectively attenuated cyst formation in vivo. This in vivo organoid model of PKD will enhance our capability to discover novel disease mechanisms and validate candidate drugs for clinical translation.

JTD Keywords: Adenylate kinase, Adult, Animal cell, Animal experiment, Animal model, Animal tissue, Article, Autophagosome, Autophagy, Autophagy (cellular), Autosomal-dominant, Calcium homeostasis, Cilia, Cilium, Cohort analysis, Controlled study, Cyclic amp, Disease, Dominant polycystic kidney, Enzyme linked immunosorbent assay, Epithelium, Exon, Expression, Female, Food and drug administration, Framework, Generation, Growth, Hepatitis a virus cellular receptor 1, Human, Human cell, Humans, Immunohistochemistry, In vitro study, In vivo study, Kidney, Kidney organoid, Kidney polycystic disease, Male, Minoxidil, Mouse, Mutations, Nonhuman, Organoid, Organoids, Platelet derived growth factor beta receptor, Pluripotent stem-cells, Polycystic kidney diseases, Protein kinase lkb1, Renin, Sequestosome 1, Single cell analysis, Single cell rna seq, Small nuclear rna, Tunel assay, Upregulation, Western blotting, Whole exome sequencing

Laser surface micropatterning of dental-grade zirconia (3Y-TZP) was explored with the objective of providing defined linear patterns capable of guiding bone-cell response.A nanosecond (ns-) laser was employed to fabricate microgrooves on the surface of 3Y-TZP discs, yielding three different groove periodicities (i.e., 30, 50 and 100 µm). The resulting topography and surface damage were characterized by confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). X-Ray diffraction (XRD) and Raman spectroscopy techniques were employed to assess the hydrothermal degradation resistance of the modified topographies. Preliminary biological studies were conducted to evaluate adhesion (6 h) of human mesenchymal stem cells (hMSC) to the patterns in terms of cell number and morphology. Finally, Staphylococcus aureus adhesion (4 h) to the microgrooves was investigated.The surface analysis showed grooves of approximately 1.8 µm height that exhibited surface damage in the form of pile-up at the edge of the microgrooves, microcracks and cavities. Accelerated aging tests revealed a slight decrease of the hydrothermal degradation resistance after laser patterning, and the Raman mapping showed the presence of monoclinic phase heterogeneously distributed along the patterned surfaces. An increase of the hMSC area was identified on all the microgrooved surfaces, although only the 50 µm periodicity, which is closer to the cell size, significantly favored cell elongation and alignment along the grooves. A decrease in Staphylococcus aureus adhesion was observed on the investigated micropatterns.The study suggests that linear microgrooves of 50 µm periodicity may help in promoting hMSC adhesion and alignment, while reducing bacterial cell attachment.Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.

JTD Keywords: abutment material, alumina toughened zirconia, antibacterial, bacterial adhesion, biofilm growth, cell adhesion, dental implants, hydrothermal degradation, implant surfaces, in-vitro, laser patterning, osseointegration, osteogenic differentiation, part 1, surface topography, y-tzp ceramics, Antibacterial, Antibacterials, Bacteria, Bone, Cell adhesion, Cell culture, Cells adhesion, Ceramics, Chemistry, Degradation resistance, Dental implants, Dental material, Dental materials, Dental prostheses, Human, Human mesenchymal stem cells, Humans, Hydrothermal degradation, Laser patterning, Laser surface, Lasers, Low-temperature degradation, Materials testing, Microscopy, electron, scanning, Nanosecond lasers, Osseointegration, Piles, Scanning electron microscopy, Staphylococcus aureus, Stem cells, Surface analysis, Surface damages, Surface properties, Surface property, Surface topography, Topography, Yttrium, Zirconia, Zirconium

Medical devices are crucial for patient care, yet even the best biomaterials lead to infections and unwanted activation of blood coagulation, potentially being life-threatening. While hydrophilic polymer brushes are the best coatings to mitigate these issues, their reliance on fossil raw materials underscores the urgency of bio-based alternatives. In this work, we introduce polymer brushes of a green solvent-based monomer, prohibiting protein adsorption, bacterial colonization, and blood clot formation at the same level as fossil-based polymer brushes. The polymer brushes are composed of N,N-dimethyl lactamide acrylate (DMLA), can be polymerized in a controlled manner, and show strong hydrophilicity as determined by thermodynamic analysis of the surface tension components. The contact of various challenging protein solutions results in repellency on the poly(DMLA) brushes. Furthermore, the poly(DMLA) brushes completely prevent the adhesion and colonization of Escherichia coli. Remarkably, upon blood contact, the poly(DMLA) brushes successfully prevent the formation of a fibrin network and leukocyte adhesion on the surface. While showcasing excellent antifouling properties similar to those of N-hydroxypropyl methacrylamide (HPMA) polymer brushes as one of the best antifouling coatings, the absence of hydroxyl groups prevents activation of the complement system in blood. We envision the polymer brushes to contribute to the future of hemocompatible coatings.

JTD Keywords: blood-plasma, coatings, contact, fossil, poly(2-methacryloyloxyethyl phosphorylcholine), protein adsorption, resistance, self-assembled monolayers, sulfobetaine, Surface-energy components

ASM deficiency in Niemann-Pick disease type A results in aberrant cellular accumulation of sphingomyelin, neuroinflammation, neurodegeneration, and early death. There is no available treatment because enzyme replacement therapy cannot surmount the blood-brain barrier (BBB). Nanocarriers (NCs) targeted across the BBB via transcytosis might help; yet, whether ASM deficiency alters transcytosis remains poorly characterized. We investigated this using model NCs targeted to intracellular adhesion molecule-1 (ICAM-1), transferrin receptor (TfR), or plasmalemma vesicle-associated protein-1 (PV1) in ASM-normal vs. ASM-deficient BBB models. Disease differentially changed the expression of all three targets, with ICAM-1 becoming the highest. Apical binding and uptake of anti-TfR NCs and anti-PV1 NCs were unaffected by disease, while anti-ICAM-1 NCs had increased apical binding and decreased uptake rate, resulting in unchanged intracellular NCs. Additionally, anti-ICAM-1 NCs underwent basolateral reuptake after transcytosis, whose rate was decreased by disease, as for apical uptake. Consequently, disease increased the effective transcytosis rate for anti-ICAM-1 NCs. Increased transcytosis was also observed for anti-PV1 NCs, while anti-TfR NCs remained unaffected. A fraction of each formulation trafficked to endothelial lysosomes. This was decreased in disease for anti-ICAM-1 NCs and anti-PV1 NCs, agreeing with opposite transcytosis changes, while it increased for anti-TfR NCs. Overall, these variations in receptor expression and NC transport resulted in anti-ICAM-1 NCs displaying the highest absolute transcytosis in the disease condition. Furthermore, these results revealed that ASM deficiency can differently alter these processes depending on the particular target, for which this type of study is key to guide the design of therapeutic NCs.© 2023. Controlled Release Society.

JTD Keywords: asm deficiency, blood-brain barrier, delivery, determines, drug, endocytosis, enzymes, icam-1, lysosomal storage disease, mechanisms, nanoparticles, natural-history, niemann-pick disease type a, pv-1, receptor-mediated transcytosis, trafficking, transferrin receptor, Asm deficiency, Blood-brain barrier, Blood–brain barrier, Drug carriers, Drug nanocarriers, Humans, Icam-1, Icam-1-targeted nanocarriers, Intercellular adhesion molecule-1, Lysosomal storage disease, Niemann-pick disease type a, Niemann-pick disease, type a, Niemann-pick diseases, Pv-1, Receptor-mediated transcytosis, Transferrin receptor

Naso- and oropharyngeal swabs are the Center for Disease Control and Prevention (CDC) -recommended disease sampling methods for respiratory viruses. The short swabbing time for sampling by these methods may lead to variability in test results. Further, these methods are mildly invasive and can cause discomfort, tearing or gag reflexes in tested individuals. If longer sampling time is coupled with lesser patient discomfort, test reliability and patient compliance can be improved. Towards this end, we developed cationic biopolymer membranes for the electrostatic capturing of viruses in the oral cavity. Here, chemically (EDC-NHS) crosslinked uncharged chitosan (CS) nanofiber membranes were conferred either with negative surface charge by anionic poly-aspartic acid (pAsp) coating or positive charge by cationic poly-L-lysine (PLL). Consistent with our preliminary findings of dynamic light scattering (DLS) size measurements showing large agglomerates of anionic virus-like particles (VLPs) and cationic PLL in solution, a 75% increase in VLP adsorption by PLL coated CS membranes was recorded by enzyme linked immunosorbent assay (ELISA), in comparison to untreated controls. It is envisaged that the electrostatic concentration of respiratory viruses on cationic membranes can be superior alternatives to traditional swabbing in the oral cavity.

JTD Keywords: Cationic biopolymer membranes, Disease sampling, Dynamic light scattering (dls), Electrostatic capture of viruses, Enzyme linked immunosorbent assay (elisa), Magnetic beads, Virus -like particles (vlps)

The combination of a powerful and broadly applicable nuclear hyperpolarization technique with emerging (near-)zero-field modalities offers novel opportunities in a broad range of nuclear magnetic resonance spectroscopy and imaging applications, including biomedical diagnostics, monitoring catalytic reactions within metal reactors and many others. These are discussed along with a roadmap for future developments.

JTD Keywords: Couplings, Hyperpolarization, Nmr, Parahydrogen, Phase, Radicals, Time

Herein, an innovative self- and pressure-adhesive biomedical implant was developed. Tissue adhesion was achieved with a thermosensitive hydrogel based on poly-(N-isopropylacrylamide-co-acrylamide), PNIPAAm-co-PAAm, grafted on a substrate composed of knitted fibers of isotactic polypropylene mesh (PP), used as surgical mesh implants. The in vitro studies, carried out with porcine skin, showed an important role of the inclusion of acrylamide-based comonomer (AAm) in the thermosensitive hydrogel PNIPAAm matrix. The bonding, peeling, and shearing energies obtained for PNIPAAm-co-PAAm increased exponentially up to three, two, and six times, respectively, compared to the gel without AAm. The physisorption and mechanical interlocking mechanisms are responsible for such improvement due to the simultaneous creation of hydrophobic and hydrophilic interactions of the thermosensitive hydrogel at temperatures higher than the lower critical solution temperature (LCST), with the porcine tissue. In addition, our bioadhesives present excellent interfacial toughness (similar to 100 J/m(2)) when compared to commercial bioglues (similar to 50 J/m(2) or lower). The results obtained represent a very promising adhesive material that is extensible to other medical devices that require atraumatic fixation to avoid chronic pain related to other fixation approaches.

JTD Keywords: Bioadhesive, Complications, Hernia-repair, Interface adhesion, Mechanicalinterlocking, Physisorption, Poly(n-isopropylacrylamide), Polypropylene mesh, Surgicalmesh, Thermosensitive hydrogel

The mechanical properties of the extracellular matrix dictate tissue behaviour. In epithelial tissues, laminin is a very abundant extracellular matrix component and a key supporting element. Here we show that laminin hinders the mechanoresponses of breast epithelial cells by shielding the nucleus from mechanical deformation. Coating substrates with laminin-111-unlike fibronectin or collagen I-impairs cell response to substrate rigidity and YAP nuclear localization. Blocking the laminin-specific integrin β4 increases nuclear YAP ratios in a rigidity-dependent manner without affecting the cell forces or focal adhesions. By combining mechanical perturbations and mathematical modelling, we show that β4 integrins establish a mechanical linkage between the substrate and keratin cytoskeleton, which stiffens the network and shields the nucleus from actomyosin-mediated mechanical deformation. In turn, this affects the nuclear YAP mechanoresponses, chromatin methylation and cell invasion in three dimensions. Our results demonstrate a mechanism by which tissues can regulate their sensitivity to mechanical signals.© 2023. The Author(s).

JTD Keywords: actin, cell migration, filaments, force transmission, localization, membrane, motility, proteins, yap, Cell adhesion, Cytoskeleton, Extracellular matrix, Fibronectins, Integrin alpha-6-beta-4, Integrins, Keratins, Laminin

The last decade has witnessed great progress in the field of adoptive cell therapies, with the authorization of Kymriah (tisagenlecleucel) in 2017 by the Food and Drug Administration (FDA) as a crucial stepstone. Since then, five more CAR-T therapies have been approved for the treatment of hematological malignancies. While this is a great step forward to treating several types of blood cancers, CAR-T cell therapies are still associated with severe side-effects such as Graft-versus-Host Disease (GvHD), cytokine release syndrome (CRS) and neurotoxicity. Because of this, there has been continued interest in Natural Killer cells which avoid these side-effects while offering the possibility to generate allogeneic cell therapies. Similar to T-cells, NK cells can be genetically modified to improve their therapeutic efficacy in a variety of ways. In contrast to T cells, viral transduction of NK cells remains inefficient and induces cytotoxic effects. Viral vectors also require a lengthy and expensive product development process and are accompanied by certain risks such as insertional mutagenesis. Therefore, non-viral transfection technologies are avidly being developed aimed at addressing these shortcomings of viral vectors. In this review we will present an overview of the potential of NK cells in cancer immunotherapies and the non-viral transfection technologies that have been explored to engineer them.Copyright © 2023 Elsevier Inc. All rights reserved.

JTD Keywords: adoptive cell therapy, cancer immunotherapy, immunotherapy, messenger-rna delivery, nanoparticle, nk cells, non -viral engineering, sonoporation, t-cell, transfection, ultrasound, Adoptive cell therapy, Cancer immunotherapy, Cell engineering, Natural-killer-cells, Nk cells, Non-viral engineering

Recent experimental evidence indicates a role for the intermediate filament vimentin in regulating cellular mechanical homeostasis, but its precise contribution remains to be discovered. Mechanical homeostasis requires a balanced bi-directional interplay between the cell's microenvironment and the cellular morphological and mechanical state-this balance being regulated via processes of mechanotransduction and mechanoresponse, commonly referred to as mechanoreciprocity. Here, we systematically analyze vimentin-expressing and vimentin-depleted cells in a swatch of in vitro cellular microenvironments varying in stiffness and/or ECM density. We find that vimentin-expressing cells maintain mechanical homeostasis by adapting cellular morphology and mechanics to micromechanical changes in the microenvironment. However, vimentin-depleted cells lose this mechanoresponse ability on short timescales, only to reacquire it on longer time scales. Indeed, we find that the morphology and mechanics of vimentin-depleted cell in stiffened microenvironmental conditions can get restored to the homeostatic levels of vimentin-expressing cells. Additionally, we observed vimentin-depleted cells increasing collagen matrix synthesis and its crosslinking, a phenomenon which is known to increase matrix stiffness, and which we now hypothesize to be a cellular compensation mechanism for the loss of vimentin. Taken together, our findings provide further insight in the regulating role of intermediate filament vimentin in mediating mechanoreciprocity and mechanical homeostasis.© 2023. The Author(s).

JTD Keywords: contributes, dynamics, focal adhesions, forces, mechanotransduction, migration, motility, organization, tissue, Intermediate-filaments

Clathrin-mediated endocytosis (CME) is an essential cellular process, conserved among eukaryotes. Yeast constitutes a powerful genetic model to dissect the complex endocytic machinery, yet there is a lack of specific pharmacological agents to interfere with CME in these organisms. TL2 is a light-regulated peptide inhibitor targeting the AP2-β-adaptin/β-arrestin interaction and that can photocontrol CME with high spatiotemporal precision in mammalian cells. Here, we study endocytic protein dynamics by live-cell imaging of the fluorescently tagged coat-associated protein Sla1-GFP, demonstrating that TL2 retains its inhibitory activity in S. cerevisiae spheroplasts. This is despite the β-adaptin/β-arrestin interaction not being conserved in yeast. Our data indicate that the AP2 α-adaptin is the functional target of activated TL2. We identified as interacting partners for the α-appendage, the Eps15 and epsin homologues Ede1 and Ent1. This demonstrates that endocytic cargo loading and sensing can be executed by conserved molecular interfaces, regardless of the proteins involved.© 2023 The Author(s).

JTD Keywords: adapters, alpha-appendage, azobenzene, cross-linker, mechanism, peptides, proteins, receptor, trafficking, Actin polymerization, Biochemistry, Biological sciences, Cell biology, Molecular biology, Natural sciences

Gas-Chromatography coupled to Ion Mobility Spectrometry (GC-IMS) based metabolomics is an emerging technique for obtaining fast, reliable untargeted metabolic fingerprints of biofluids. The generated raw data is highly dimensional and complex, suffers from baseline problems, misalignments, long peak tails and strong nonlinearities that must be corrected to extract chemically relevant features from samples. In this work, we present our GCIMS R package, which includes spectra loading, metadata handling, denoising, baseline correction, spectral and chromatographic alignment, peak detection, integration, and peak clustering to produce a peak table ready for multivariate data analysis. We discuss package design decisions, and, for illustration purposes, we show a case study of sex discrimination on the basis of the volatile compounds in urine samples. The GCIMS package provides a user-friendly workflow for non-code developers to process their raw data samples.

JTD Keywords: Breath analysis, Headspace, Ims, Urine

The formation of a protein corona, where proteins spontaneously adhere to the surface of nanomaterials in biological environments, leads to changes in their physicochemical properties and subsequently affects their intended biomedical functionalities. Most current methods to study protein corona formation are ensemble-averaging and either require fluorescent labeling, washing steps, or are only applicable to specific types of particles. Here we introduce real-time all-optical nanoparticle analysis by scattering microscopy (RONAS) to track the formation of protein corona in full serum, at the single-particle level, without any labeling. RONAS uses optical scattering microscopy and enables real-time and in situ tracking of protein adsorption on metallic and dielectric nanoparticles with different geometries directly in blood serum. We analyzed the adsorbed protein mass, the affinity, and the kinetics of the protein adsorption at the single particle level. While there is a high degree of heterogeneity from particle to particle, the predominant factor in protein adsorption is surface chemistry rather than the underlying nanoparticle material or size. RONAS offers an in-depth understanding of the mechanisms related to protein coronas and, thus, enables the development of strategies to engineer efficient bionanomaterials.

JTD Keywords: Dielectric nanoparticles, Optical microscopy, Plasmonic nanoparticles, Protein corona, Protein corona,plasmonic nanoparticles,dielectricnanoparticles,optical microscopy,single particle, Silica nanoparticles,gold nanoparticles,surface-charge,adsorption,albumin,binding,size,transferrin,mas, Single particles

Adherens junctions (AJs) allow cell contact to inhibit epithelial migration yet also permit epithelia to move as coherent sheets. How, then, do cells identify which contacts will inhibit locomotion? Here, we show that in human epithelial cells this arises from the orientation of cortical flows at AJs. When the leader cells from different migrating sheets make head-on contact with one another, they assemble AJs that couple together oppositely directed cortical flows. This applies a tensile signal to the actin-binding domain (ABD) of a-cate-nin, which provides a clutch to promote lateral adhesion growth and inhibit the lamellipodial activity neces-sary for migration. In contrast, AJs found between leader cells in the same migrating sheet have cortical flows aligned in the same direction, and no such mechanical inhibition takes place. Therefore, a-catenin mechano-sensitivity in the clutch between E-cadherin and cortical F-actin allows cells to interpret the direction of motion via cortical flows and signal for contact to inhibit locomotion.

JTD Keywords: Clutch, Contact inhibition of locomotion, Cortical flow, E-cadherin adhesion, Mechanical tension, Α-catenin

The incorporation of exogenous lactate into cardiac tissues is a regenerative strategy that is rapidly gaining attention. In this work, two polymeric platforms were designed to achieve a sustained release of lactate, combining immediate and prolonged release profiles. Both platforms contained electrospun poly(lactic acid) (PLA) fibers and an alginate (Alg) hydrogel. In the first platform, named L/K(x)/Alg-PLA, lactate and proteinase K (x mg of enzyme per 1 g of PLA) were directly loaded into the Alg hydrogel, into which PLA fibers were assembled. In the second platform, L/Alg-K(x)/PLA, fibers were produced by electrospinning a proteinase K:PLA solution and, subsequently, assembled within the lactate-loaded hydrogel. After characterizing the chemical, morphological, and mechanical properties of the systems, as well as their cytotoxicity, the release profiles of the two platforms were determined considering different amounts of proteinase K (x = 5.2, 26, and 52 mg of proteinase K per 1 g of PLA), which is known to exhibit a broad cleavage activity. The profiles obtained using L/Alg-K(x)/PLA platforms with x = 26 and 52 were the closest to the criteria that must be met for cardiac tissue regeneration. Finally, the amount of lactate directly loaded in the Alg hydrogel for immediate release and the amount of protein in the electrospinning solution were adapted to achieve a constant lactate release of around 6 mM per day over 1 or 2 weeks. In the optimized bioplatform, in which 6 mM lactate was loaded in the hydrogel, the amount of fibers was increased by a factor of x3, the amount of enzyme was adjusted to 40 mg per 1 g of PLA, and a daily lactate release of 5.9 +/- 2.7 mM over a period of 11 days was achieved. Accordingly, the engineered device fully satisfied the characteristics and requirements for heart tissue regeneration.

JTD Keywords: biodegradable fibers, cardiac tissue regeneration, cell, drug-release, elastic-modulus, electrospinning, heart, nanoindentation, plasma treatment, proteinase, scaffold, stiffness, Alginate, Alginates, Biodegradable fibers, Cardiac tissue, Cardiac tissue regeneration, Cell, Delayed-action preparations, Drug-release, Elastic-modulus, Electrospinning, Endopeptidase k, Heart, Hydrogels, Lactic acid, Nanoindentation, Plasma treatment, Poly(lactide), Polyesters, Proteinase, Regeneration, Scaffold, Skeletal-muscle, Stiffness

Prior studies demonstrated that encapsulation in poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) enhanced the delivery of enzymes used for replacement therapy (ERT) of lysosomal storage disorders (LSDs). This study examined how the copolymer lactide:glycolide ratio impacts encapsulation, physicochemical characteristics, stability, and release under lysosomal conditions. Hyaluronidase, deficient in mucopolysaccharidosis IX, was encapsulated in NPs synthesized using 50:50, 60:40, or 75:25 lactide:glycolide copolymers. All NPs had diameters compatible with cellular transport (≤168 nm) and polydispersity indexes (≤0.16) and ζ-potentials (≤-35 mV) compatible with colloidal stability. Yet, their encapsulation efficiency varied, with 75:25 NPs and 60:40 NPs having the lowest and highest EE, respectively (15% vs. 28%). Under lysosomal conditions, the 50:50 copolymer degraded fastest (41% in 1 week), as expected, and the presence of a targeting antibody coat did not alter this result. Additionally, 60:40 NPs destabilized fastest (<1 week) because of their smaller diameter, and 75:25 NPs did not destabilize in 4 weeks. All formulations presented burst release under lysosomal conditions (56-78% of the original load within 30 min), with 50:50 and 60:40 NPs releasing an additional small fraction after week 1. This provided 4 weeks of sustained catalytic activity, sufficient to fully degrade a substrate. Altogether, the 60:40 NP formulation is preferred given its higher EE, and 50:50 NPs represent a valid alternative, while the highest stability of 75:25 NPs may impair lysosomes. These results can guide future studies aiming to translate PLGA NP-based ERT for this and other LSDs.

JTD Keywords: biodegradation, copolymer ratio, degradation, drug-delivery, emulsification, enzyme release, enzyme replacement therapy, hyaluronidase, mechanisms, microspheres, nanoparticle stability, poly(lactide-co-glycolide) nanoparticles, size, sphingomyelinase, transport, Central-nervous-system, Copolymer ratio, Enzyme release, Enzyme replacement therapy, Hyaluronidase, Lysosomal storage disorder, Nanoparticle stability, Poly(lactide-co-glycolide) nanoparticles

The insect Galleria mellonella is an alternative animal model widely used for studying bacterial infections. It presents a wide range of advantages, including its low cost, easy maintenance and lack of ethical constraints. Among other features, their innate immune system is very similar to that of mammals. In this study, we dissected several larvae infected with important human pathogens: Mycobacterium abscessus, Staphylococcus aureus and Pseudomonas aeruginosa. By observing the fat body, gut, trachea, and hemolymph under the microscope, we were able to describe where bacteria tend to disseminate. We also quantified the number of bacteria in the hemolymph throughout the infection course and found significant differences between the different pathogens. With this work, we aimed to better understand the behavior and dissemination of bacteria in the infected larvae.Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.

JTD Keywords: Bacteria, Dissection, Galleria mellonella, Infection

Artemisinin, curcumin or quercetin, alone or in combination, were loaded in nutriosomes, special phospholipid vesicles enriched with Nutriose FM06®, a soluble dextrin with prebiotic activity, that makes these vesicles suitable for oral delivery. The resulting nutriosomes were sized between 93 and 146 nm, homogeneously dispersed, and had slightly negative zeta potential (around -8 mV). To improve their shelf life and storability over time, vesicle dispersions were freeze-dried and stored at 25 °C. Results confirmed that their main physico-chemical characteristics remained unchanged over a period of 12 months. Additionally, their size and polydispersity index did not undergo any significant variation after dilution with solutions at different pHs (1.2 and 7.0) and high ionic strength, mimicking the harsh conditions of the stomach and intestine. An in vitro study disclosed the delayed release of curcumin and quercetin from nutriosomes (∼53% at 48 h) while artemisinin was quickly released (∼100% at 48 h). Cytotoxicity assays using human colon adenocarcinoma cells (Caco-2) and human umbilical vein endothelial cells (HUVECs) proved the high biocompatibility of the prepared formulations. Finally, in vitro antimalarial activity tests, assessed against the 3D7 strain of Plasmodium falciparum, confirmed the effectiveness of nutriosomes in the delivery of curcumin and quercetin, which can be used as adjuvants in the antimalaria treatment. The efficacy of artemisinin was also confirmed but not improved. Overall results proved the possible use of these formulations as an accompanying treatment of malaria infections.Copyright © 2023. Published by Elsevier B.V.

JTD Keywords: artemisinin, delivery, flavonol, formulations, liposomes, malaria infections, nanomedicine, nutriose (r) fm06, oral administration, plasmodium falciparum, In-vitro, Liposomes, Malaria infections, Nanomedicine, Nutriose® fm06, Oral administration, Plasmodium falciparum

We aimed to characterize the cerebral hemodynamic response to obstructive sleep apnea/hypopnea events, and evaluate their association to polysomnographic parameters. The characterization of the cerebral hemodynamics in obstructive sleep apnea (OSA) may add complementary information to further the understanding of the severity of the syndrome beyond the conventional polysomnography.Severe OSA patients were studied during night sleep while monitored by polysomnography. Transcranial, bed-side diffuse correlation spectroscopy (DCS) and frequency-domain near-infrared diffuse correlation spectroscopy (NIRS-DOS) were used to follow microvascular cerebral hemodynamics in the frontal lobes of the cerebral cortex. Changes in cerebral blood flow (CBF), total hemoglobin concentration (THC), and cerebral blood oxygen saturation (StO2) were analyzed.We considered 3283 obstructive apnea/hypopnea events from sixteen OSA patients (Age (median, interquartile range) 57 (52-64.5); females 25%; AHI (apnea-hypopnea index) 84.4 (76.1-93.7)). A biphasic response (maximum/minimum followed by a minimum/maximum) was observed for each cerebral hemodynamic variable (CBF, THC, StO2), heart rate and peripheral arterial oxygen saturation (SpO2). Changes of the StO2 followed the dynamics of the SpO2, and were out of phase from the THC and CBF. Longer events were associated with larger CBF changes, faster responses and slower recoveries. Moreover, the extrema of the response to obstructive hypopneas were lower compared to apneas (p < .001).Obstructive apneas/hypopneas cause profound, periodic changes in cerebral hemodynamics, including periods of hyper- and hypo-perfusion and intermittent cerebral hypoxia. The duration of the events is a strong determinant of the cerebral hemodynamic response, which is more pronounced in apnea than hypopnea events.© The Author(s) 2023. Published by Oxford University Press on behalf of Sleep Research Society.

JTD Keywords: cerebral hemodynamics, desaturation, diffuse correlation spectroscopy, duration, hypopnea, hypoxemia, near-infrared spectroscopy, optical pathlength, oxygenation, severity, sleep disorder, spectroscopy, tissue, Adult, Airway obstruction, Apnea hypopnea index, Arterial oxygen saturation, Article, Blood oxygen tension, Blood-flow, Brain blood flow, Brain cortex, Cerebral hemodynamics, Controlled study, Diffuse correlation spectroscopy, Disease severity, Female, Frequency, Frontal lobe, Heart rate, Hemodynamics, Hemoglobin, Hemoglobin determination, Human, Humans, Major clinical study, Male, Near infrared spectroscopy, Near-infrared spectroscopy, Obstructive sleep apnea, Oxygen, Periodicity, Polysomnography, Sleep apnea syndromes, Sleep apnea, obstructive, Sleep disorder, Spectroscopy, near-infrared

Ratiometric fluorescent nanothermometers with near-infrared emission play an important role in in vivo sensing since they can be used as intracellular thermal sensing probes with high spatial resolution and high sensitivity, to investigate cellular functions of interest in diagnosis and therapy, where current approaches are not effective. Herein, the temperature-dependent fluorescence of organic nanoparticles is designed, synthesized, and studied based on the dual emission, generated by monomer and excimer species, of the tris(2,4,6-trichlorophenyl)methyl radical (TTM) doping organic nanoparticles (TTMd-ONPs), made of optically neutral tris(2,4,6-trichlorophenyl)methane (TTM-αH), acting as a matrix. The excimer emission intensity of TTMd-ONPs decreases with increasing temperatures whereas the monomer emission is almost independent and can be used as an internal reference. TTMd-ONPs show a great temperature sensitivity (3.4% K-1 at 328 K) and a wide temperature response at ambient conditions with excellent reversibility and high colloidal stability. In addition, TTMd-ONPs are not cytotoxic and their ratiometric outputs are unaffected by changes in the environment. Individual TTMd-ONPs are able to sense temperature changes at the nano-microscale. In vivo thermometry experiments in Caenorhabditis elegans (C. elegans) worms show that TTMd-ONPs can locally monitor internal body temperature changes with spatio-temporal resolution and high sensitivity, offering multiple applications in the biological nanothermometry field.© 2023 The Authors. Small published by Wiley-VCH GmbH.

JTD Keywords: dual emission, elegans, excimer emission, fluorescence, in vivo sensing, luminescence, nanoparticles, organic radical nanoparticles, ratiometric nanothermometers, sensors, thermometry, trityl radicals, Caenorhabditis elegans, Excimer emission, In vivo sensing, Intracellular ph, Luminescence, Organic radical nanoparticles, Ratiometric nanothermometers, Trityl radicals

Live cell actin visualization is fundamental for exploring cellular motility, cytokinesis, intracellular transport, and other correlated functions. The current imaging techniques that allow imaging of actin in its native environment are optical and electron microscopy. Such imaging techniques offer high enough resolution to investigate the ultrastructure of actin however they come at the expense of actin integrity. Inspired by the lack of suitable probes that preserve actin's integrity, we designed a cyclometalated Ir (III) complex that interacts with live cells and displays light switch behaviour upon specific actin binding. The exceptional photophysical properties of the proposed probe allow unprecedented resolution of cytoskeleton ultrastructures under stimulated emission depletion (STED) super-resolution nanoscopy. Moreover, the Ir complex enables the capability of visualizing actin polymers and periodicity under correlative light electron microscopy (CLEM) and liquid-phase electron microscopy (LPEM) at similar to 8 nm resolution.

JTD Keywords: Actin dynamics, Actin targeting, Adhesion, Cells, Clem, Fluorescent, Iridium (iii) complex, Lead, Light, Lpem, Super-resolution ultrastructures

Photochemically induced dynamic nuclear polarization (photo-CIDNP) enables nuclear spin ordering by irradiating samples with light. Polarized spins are conventionally detected via high-field chemical-shift-resolved NMR (above 0.1 T). In this Letter, we demonstrate in situ low-field photo-CIDNP measurements using a magnetically shielded fast-field-cycling NMR setup detecting Larmor precession via atomic magnetometers. For solutions comprising mM concentrations of the photochemically polarized molecules, hyperpolarized 1H magnetization is detected by pulse-acquired NMR spectroscopy. The observed NMR line widths are about 5 times narrower than normally anticipated in high-field NMR and are systematically affected by light irradiation during the acquisition period, reflecting a reduction of the transverse relaxation time constant, T2*, on the order of 10%. Magnetometer-detected photo-CIDNP spectroscopy enables straightforward observation of spin-chemistry processes in the ambient field range from a few nT to tens of mT. Potential applications of this measuring modality are discussed.

JTD Keywords: field-dependence, mechanism, nmr, parahydrogen, photo-cidnp, polarization, quinone, spin-hyperpolarization, Radical-pair

Ex vivo-loaded white blood cells (WBC) can transfer cargo to pathological foci in the central nervous system (CNS). Here we tested affinity ligand driven in vivo loading of WBC in order to bypass the need for ex vivo WBC manipulation. We used a mouse model of acute brain inflammation caused by local injection of tumor necrosis factor alpha (TNF-α). We intravenously injected nanoparticles targeted to intercellular adhesion molecule 1 (anti-ICAM/NP). We found that (A) at 2 h, >20% of anti-ICAM/NP were localized to the lungs; (B) of the anti-ICAM/NP in the lungs >90% were associated with leukocytes; (C) at 6 and 22 h, anti-ICAM/NP pulmonary uptake decreased; (D) anti-ICAM/NP uptake in brain increased up to 5-fold in this time interval, concomitantly with migration of WBCs into the injured brain. Intravital microscopy confirmed transport of anti-ICAM/NP beyond the blood-brain barrier and flow cytometry demonstrated complete association of NP with WBC in the brain (98%). Dexamethasone-loaded anti-ICAM/liposomes abrogated brain edema in this model and promoted anti-inflammatory M2 polarization of macrophages in the brain. In vivo targeted loading of WBC in the intravascular pool may provide advantages of coopting WBC predisposed to natural rapid mobilization from the lungs to the brain, connected directly via conduit vessels.

JTD Keywords: drug delivery, icam-1, inflammation, lung injury, messenger-rna, migration, model, nanoparticles, neutrophils, pharmacokinetics, t-cells, white bloodcells, Adhesion molecules, Brain, Drug delivery, Inflammation, Nanoparticles, Pharmacokinetics, White blood cells

The function of organs such as lungs, kidneys and mammary glands relies on the three-dimensional geometry of their epithelium. To adopt shapes such as spheres, tubes and ellipsoids, epithelia generate mechanical stresses that are generally unknown. Here we engineer curved epithelial monolayers of controlled size and shape and map their state of stress. We design pressurized epithelia with circular, rectangular and ellipsoidal footprints. We develop a computational method, called curved monolayer stress microscopy, to map the stress tensor in these epithelia. This method establishes a correspondence between epithelial shape and mechanical stress without assumptions of material properties. In epithelia with spherical geometry we show that stress weakly increases with areal strain in a size-independent manner. In epithelia with rectangular and ellipsoidal cross-section we find pronounced stress anisotropies that impact cell alignment. Our approach enables a systematic study of how geometry and stress influence epithelial fate and function in three-dimensions.© 2023. The Author(s).

JTD Keywords: cell, forces, morphogenesis, tension, E-cadherin, Epithelial cells, Epithelium, Microscopy, Stress, mechanical

Fibrillar collagens are the most abundant extracellular matrix components in non-small cell lung cancer (NSCLC). However, the potential of collagen fiber descriptors as a source of clinically relevant biomarkers in NSCLC is largely unknown. Similarly, our understanding of the aberrant collagen organization and associated tumor-promoting effects is very scarce. To address these limitations, we identified a digital pathology approach that can be easily implemented in pa-thology units based on CT-FIRE software (version 2; https://loci.wisc.edu/software/ctfire) analysis of Picrosirius red (PSR) stains of fibrillar collagens imaged with polarized light (PL). CT-FIRE set-tings were pre-optimized to assess a panel of collagen fiber descriptors in PSR-PL images of tissue microarrays from surgical NSCLC patients (106 adenocarcinomas [ADC] and 89 squamous cell carcinomas [SCC]). Using this approach, we identified straightness as the single high-accuracy diagnostic collagen fiber descriptor (average area under the curve 1/4 0.92) and fiber density as the single descriptor consistently associated with a poor prognosis in both ADC and SCC inde-pendently of the gold standard based on the TNM staging (hazard ratio, 2.69; 95% CI, 1.55-4.66; P < .001). Moreover, we found that collagen fibers were markedly straighter, longer, and more aligned in tumor samples compared to paired samples from uninvolved pulmonary tissue, particularly in ADC, which is indicative of increased tumor stiffening. Consistently, we observed an increase in a panel of stiffness-associated processes in the high collagen fiber density patient group selectively in ADC, including venous/lymphatic invasion, fibroblast activation (a-smooth muscle actin), and immune evasion (programmed death-ligand 1). Similarly, a transcriptional correlation analysis supported the potential involvement of the major YAP/TAZ pathway in ADC. Our results provide a proof-of-principle to use CT-FIRE analysis of PSR-PL images to assess new collagen fiber-based diagnostic and prognostic biomarkers in pathology units, which may improve the clinical management of patients with surgical NSCLC. Our findings also unveil an aberrant stiff micro -environment in lung ADC that may foster immune evasion and dissemination, encouraging future work to identify therapeutic opportunities. (c) 2023 THE AUTHORS. Published by Elsevier Inc. on behalf of the United States & Canadian Academy of Pathology. This is an open access article under the CC BY-NC-ND license (http://creativecommo ns.org/licenses/by-nc-nd/4.0/).

JTD Keywords: biomarkers, collagen, ct-fire, lung cancer, mechanobiology, Adenocarcinoma, Association, Biomarkers, Collagen, Ct-fire, Differentiation, Expression, Extracellular-matrix, I collagen, Invasion, Lung cancer, Mechanobiology, Microenvironment, Signature, Survival, Tumor microenvironment

Previous studies have shown hydrophilic/hydrophobic implant surfaces stimulate/hinder osseointegration. An analogous concept was applied here using common biological functional groups on a model surface to promote oral keratinocytes (OKs) proliferation and hemidesmosomes (HD) to extend implant lifespans through increased soft tissue attachment. However, it is unclear what physicochemistry stimulates HDs. Thus, common biological functional groups (NH2 , OH, and CH3 ) were functionalized on glass using silanization. Non-functionalized plasma-cleaned glass and H silanization were controls. Surface modifications were confirmed with X-ray photoelectron spectroscopy and water contact angle. The amount of bovine serum albumin (BSA) and fibrinogen, and BSA thickness, were assessed to understand how adsorbed protein properties were influenced by physicochemistry and may influence HDs. OKs proliferation was measured, and HDs were quantified with immunofluorescence for collagen XVII and integrin β4. Plasma-cleaned surfaces were the most hydrophilic group overall, while CH3 was the most hydrophobic and OH was the most hydrophilic among functionalized groups. Modification with the OH chemical group showed the highest OKs proliferation and HD expression. The OKs response on OH surfaces appeared to not correlate to the amount or thickness of adsorbed model proteins. These results reveal relevant surface physicochemical features to favor HDs and improve implant soft tissue attachment.© 2023 The Authors. Journal of Biomedical Materials Research Part A published by Wiley Periodicals LLC.

JTD Keywords: attachment, chemistry, collagen, differentiation, epithelial-cells, hemidesmosome, implant, in-vitro, integrin, keratinocyte, mechanism, organosilane, physicochemistry, protein adsorption, Attachment, Cell-adhesion, Physicochemistry

This work presents a novel nanoparticle-based thermosensor implant able to reveal the precise temperature variations along the polymer filaments, as it contracts and expands due to changes in the macroscale local temperature. The multimodal device is able to trace the position and the temperature of a polypropylene mesh, employed in abdominal hernia repair, by combining plasmon resonance and Raman spectroscopy with hydrogel responsive system. The novelty relies on the attachment of the biocompatible nanoparticles, based on gold stabilized by a chitosan-shell, already charged with the Raman reporter (RaR) molecules, to the robust prosthesis, without the need of chemical linkers. The SERS enhanced effect observed is potentiated by the presence of a quite thick layer of the copolymer (poly(N-isopropylacrylamide)-co-poly(acrylamide)) hydrogel. At temperatures above the LCST of PNIPAAm-co-PAAm, the water molecules are expulsed and the hydrogel layer contracts, leaving the RaR molecules more accessible to the Raman source. In vitro studies with fibroblast cells reveal that the functionalized surgical mesh is biocompatible and no toxic substances are leached in the medium. The mesh sensor opens new frontiers to semi-invasive diagnosis and infection prevention in hernia repair by using SERS spectroscopy. It also offers new possibilities to the functionalization of other healthcare products.© 2023 Wiley-VCH GmbH.

JTD Keywords: adhesion, blends, chitosan, gold nanoparticles, poly(n-isopropylacrylamide), polypropylene mesh, polypropylene meshes, repair, scattering, silver, surgical implants, thermosensitive hydrogels, toxicity, Chitosan, Gold nanoparticles, Polypropylene meshes, Surgical implants, Thermosensitive hydrogels

Autonomous circadian clocks exist in nearly every mammalian cell type. These cellular clocks are subjected to a multilayered regulation sensitive to the mechanochemical cell microenvironment. Whereas the biochemical signaling that controls the cellular circadian clock is increasingly well understood, mechanisms underlying regulation by mechanical cues are largely unknown. Here we show that the fibroblast circadian clock is mechanically regulated through YAP/TAZ nuclear levels. We use high-throughput analysis of single-cell circadian rhythms and apply controlled mechanical, biochemical, and genetic perturbations to study the expression of the clock gene Rev-erbα. We observe that Rev-erbα circadian oscillations are disrupted with YAP/TAZ nuclear translocation. By targeted mutations and overexpression of YAP/TAZ, we show that this mechanobiological regulation, which also impacts core components of the clock such as Bmal1 and Cry1, depends on the binding of YAP/TAZ to the transcriptional effector TEAD. This mechanism could explain the impairment of circadian rhythms observed when YAP/TAZ activity is upregulated, as in cancer and aging.© 2023 Abenza et al.

JTD Keywords: activation, dynamics, forces, growth, hippo pathway, liver, platform, time, transcription, Animals, Circadian clocks, Circadian rhythm, Gene-expression, Mammals, Signal transduction, Tea domain transcription factors, Transcriptional coactivator with pdz-binding motif proteins, Yap-signaling proteins

The functionalization of 3D-printed poly-l-lactic acid (PLLA) and poly(l-lactic-co-ε-caprolactone) (PLCL) bioresorbable stents has been successfully achieved with linear RGDS and YIGSR peptides, as well as a dual platform containing both motifs within a single biomolecule.

JTD Keywords: adsorbed fibrinogen, chemistry, endothelialization, immobilization, platelets, plla, selectivity, surface, titanium, Absorbable implants, Cell adhesion, Endothelial cells, In-vitro hemocompatibility, Peptides, Polymers, Printing, three-dimensional, Stents, Tyrosyl-isoleucyl-glycyl-seryl-arginine

JTD Keywords: 3d printing, Biodegradable, Guided bone regeneration, Implant

Titanium implantation success may be compromised by Staphylococcus aureus surface colonization and posterior infection. To avoid this issue, different strategies have been investigated to promote an antibacterial character to titanium. In this work, two antibacterial agents (silver nanoparticles and a multifunctional antimicrobial peptide) were used to coat titanium surfaces. The modulation of the nanoparticle (≈32.1 ± 9.4 nm) density on titanium could be optimized, and a sequential functionalization with both agents was achieved through a two-step functionalization method by means of surface silanization. The antibacterial character of the coating agents was assessed individually as well as combined. The results have shown that a reduction in bacteria after 4 h of incubation can be achieved on all the coated surfaces. After 24 h of incubation, however, the individual antimicrobial peptide coating was more effective than the silver nanoparticles or their combination against Staphylococcus aureus. All tested coatings were non-cytotoxic for eukaryotic cells.

JTD Keywords: antimicrobial peptide, biomaterials, bone, coatings, performance, ph, resistance, silanization, silver nanoparticles, staphylococcus aureus, Anti-bacterial agents, Antimicrobial peptide, Coated materials, biocompatible, Metal nanoparticles, Reduces bacterial adhesion, Silanization, Silver, Silver nanoparticles, Staphylococcus aureus, Surface properties, Titanium, Titanium functionalization

JTD Keywords: coverage path planning (cpp), gas distribution map (gdm), gas source localization (gsl), metal oxide semiconductor sensor, quality, robot, system, Nano-quadcopter, Source localization

This work proposes a microfibers-hydrogel assembled composite as delivery vehicle able to combine into a single system both burst and prolonged release of lactate. The prolonged release of lactate has been achieved by electrospinning a mixture of polylactic acid and proteinase K (26.0 mg of proteinase K and 0.99 g of PLA dissolved in 6 mL of 2:1 chloroform:acetone in the optimal case), which is a protease that catalyzes the degradation of polylactic acid into lactate. The degradation of microfibers into lactate reflects that proteinase K preserves its enzymatic activity even after the electrospinning process because of the mild operational conditions used. Besides, burst release is obtained from the lactate-loaded alginate hydrogel. The successful assembly between the lactate-loaded hydrogel and the polylactic acid/proteinase K fibers has been favored by applying a low-pressure (0.3 mbar at 300 W) oxygen plasma treatment, which transforms hydrophobic fibers into hydrophilic while the enzymatic activity is still maintained. The composite displays both fast (< 24 h) and sustained (> 10 days) lactate release, and allows the modulation of the release by adjusting either the amount of loaded lactate or the amount of active enzyme.Copyright © 2023. Published by Elsevier B.V.

JTD Keywords: biodegradable fibers, proteases, regeneration, repair, Alginate, Alginates, Biodegradable fibers, Endopeptidase k, Hydrogels, Lactic acid, Myocardial-infarction, Polymers, Proteases

Virtually, all implantable medical devices are susceptible to infection. As the main healthcare issue concerning implantable devices is the elevated risk of infection, different strategies based on the coating or functionalization of biomedical devices with antiseptic agents or antibiotics are proposed. In this work, an alternative approach is presented, which consists of the functionalization of implantable medical devices with sensors capable of detecting infection at very early stages through continuous monitoring of the bacteria metabolism. This approach, which is implemented in surgical sutures as a representative case of implantable devices susceptible to bacteria colonization, is expected to minimize the risk of worsening the patient's clinical condition. More specifically, non-absorbable polypropylene/polyethylene (PP/PE) surgical sutures are functionalized with conducting polymers using a combination of low-pressure oxygen plasma, chemical oxidative polymerization, and anodic polymerization, to detect metabolites coming from bacteria respiration. Functionalized suture yarns are used for real-time monitoring of bacteria growth, demonstrating the potential of this strategy to fight against infections.© 2023 Wiley-VCH GmbH.

JTD Keywords: adhesion, biofilm, conducting polymers, contamination, derivatives, detections, functionalized sutures, nadh, poly(3,4-ethylenedioxythiophene), Bacteria growth, Conducting polymers, Detections, Functionalized sutures, Monofilament, Nadh

The term 'mechanosensation' describes the capacity of cells to translate mechanical stimuli into the coordinated regulation of intracellular signals, cellular function, gene expression and epigenetic programming. This capacity is related not only to the sensitivity of the cells to tissue motion, but also to the decryption of tissue geometric arrangement and mechanical properties. The cardiac stroma, composed of fibroblasts, has been historically considered a mechanically passive component of the heart. However, the latest research suggests that the mechanical functions of these cells are an active and necessary component of the developmental biology programme of the heart that is involved in myocardial growth and homeostasis, and a crucial determinant of cardiac repair and disease. In this Review, we discuss the general concept of cell mechanosensation and force generation as potent regulators in heart development and pathology, and describe the integration of mechanical and biohumoral pathways predisposing the heart to fibrosis and failure. Next, we address the use of 3D culture systems to integrate tissue mechanics to mimic cardiac remodelling. Finally, we highlight the potential of mechanotherapeutic strategies, including pharmacological treatment and device-mediated left ventricular unloading, to reverse remodelling in the failing heart.© 2022. Springer Nature Limited.

JTD Keywords: cardiomyocyte proliferation, cross-linking, extracellular-matrix, focal adhesions, gene-expression, mechanical regulation, myocardial-infarction, substrate stiffness affects, t-cells, Ventricular assist device

The TGF-β1 transcription factor SMAD3 is epigenetically repressed in tumour-associated fibroblasts (TAFs) from lung squamous cell carcinoma (SCC) but not adenocarcinoma (ADC) patients, which elicits a compensatory increase in SMAD2 that renders SCC-TAFs less fibrotic. Here we examined the effects of altered SMAD2/3 in fibroblast migration and its impact on the desmoplastic stroma formation in lung cancer.We used a microfluidic device to examine descriptors of early protrusions and subsequent migration in 3D collagen gels upon knocking down SMAD2 or SMAD3 by shRNA in control fibroblasts and TAFs.High SMAD3 conditions as in shSMAD2 fibroblasts and ADC-TAFs exhibited a migratory advantage in terms of protrusions (fewer and longer) and migration (faster and more directional) selectively without TGF-β1 along with Erk1/2 hyperactivation. This enhanced migration was abrogated by TGF-β1 as well as low glucose medium and the MEK inhibitor Trametinib. In contrast, high SMAD2 fibroblasts were poorly responsive to TGF-β1, high glucose and Trametinib, exhibiting impaired migration in all conditions.The basal migration advantage of high SMAD3 fibroblasts provides a straightforward mechanism underlying the larger accumulation of TAFs previously reported in ADC compared to SCC. Moreover, our results encourage using MEK inhibitors in ADC-TAFs but not SCC-TAFs.© 2022. The Author(s).

JTD Keywords: cancer, cell, degradation, nintedanib, osteoblast migration, phenotype, progression, protrusion dynamics, smad3, Growth-factor-beta

Background Pilocytic astrocytoma (PA) is the most common pediatric brain tumor and a mitogen-activated protein kinase (MAPK)-driven disease. Oncogenic MAPK-signaling drives the majority of cells into oncogene-induced senescence (OIS). While OIS induces resistance to antiproliferative therapies, it represents a potential vulnerability exploitable by senolytic agents. Methods We established new patient-derived PA cell lines that preserve molecular features of the primary tumors and can be studied in OIS and proliferation depending on expression or repression of the SV40 large T antigen. We determined expression of anti-apoptotic BCL-2 members in these models and primary PA. Dependence of senescent PA cells on anti-apoptotic BCL-2 members was investigated using a comprehensive set of BH3 mimetics. Results Senescent PA cells upregulate BCL-XL upon senescence induction and show dependency on BCL-XL for survival. BH3 mimetics with high affinity for BCL-XL (BCL-XLi) reduce metabolic activity and induce mitochondrial apoptosis in senescent PA cells at nano-molar concentrations. In contrast, BH3 mimetics without BCL-XLi activity, conventional chemotherapy, and MEK inhibitors show no effect. Conclusions Our data demonstrate that BCL-XL is critical for survival of senescent PA tumor cells and provides proof-of-principle for the use of clinically available BCL-XL-dependent senolytics.

JTD Keywords: bcl-xl, bh3 mimetics, oncogene-induced senescence, Bcl-xl, Bh3 mimetics, Expression, Family, Inhibitor, Low-grade glioma, Navitoclax, Oncogene-induced senescence, Pilocytic astrocytoma, Stem-cells

Actual polymeric wound closure devices are not optimal for load-bearing applications due to the low mechanical properties and the risk of inflammation and bacterial infection mainly produced by multifil-ament and braided configurations. Biodegradable metallic Zn alloys are promising materials candidates; however, mechanical performance, corrosion behaviour, and biological response should be controlled in order to inhibit the risk of inflammation and bacterial infection. To this end, a Zn-2Ag (2 wt% Ag) alloy was processed by ECAP to evaluate the concurrent combined effect of grain refinement and Ag alloying on biodegradation and antibacterial activity. Two ECAP cycles were successfully applied to a Zn-2Ag alloy obtaining a homogeneous ultra-fine-grained structure in which nanoindentation maps suggested isotro-pic mechanical properties. Lower UTS and YS with higher elongation was reported after ECAP with similar corrosion rates as before processing. ECAP processed samples showed a homogeneous Ag+ release below the minimum inhibitory concentration for S. Aureus and no antibacterial effect was observed by diffusion. As expected, the presence of Ag in Zn-Ag alloys reduced bacterial attachment. Nevertheless, ECAP processed Zn-2Ag provided an excellent antibacterial activity after 3 h probably caused by the uniformly degraded and thus, non- stable, surface observed after bacterial adhesion.(c) 2023 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

JTD Keywords: Behavior, Binary alloys, Biodegradable zinc-alloys, Biomaterials, Equal channel angular pressing, Grain-refinement, In-vitro degradation, Mg, Microstructure, Nanoindentation, Progress, Staphylococcus-aureus, Temperature superplasticity, Ultrafine-grained materials, Zinc alloys, Zn alloys

Topographical patterns are a powerful tool to study directional migration. Grooved substrates have been extensively used as in vitro models of aligned extracellular matrix fibers because they induce cell elongation, alignment, and migration through a phenomenon known as contact guidance. This process, which involves the orientation of focal adhesions, F-actin, and microtubule cytoskeleton along the direction of the grooves, has been primarily studied on hard materials of non-physiological stiffness. But how it unfolds when the stiffness of the grooves varies within the physiological range is less known. Here we show that substrate stiffness modulates the cellular response to topographical contact guidance. We find that for fibroblasts, while focal adhesions and actin respond to topography independently of the stiffness, microtubules show a stiffness-dependent response that regulates contact guidance. On the other hand, both clusters and single breast carcinoma epithelial cells display stiffness-dependent contact guidance, leading to more directional and efficient migration when increasing substrate stiffness. These results suggest that both matrix stiffening and alignment of extracellular matrix fibers cooperate during directional cell migration, and that the outcome differs between cell types depending on how they organize their cytoskeletons.© 2023 The Authors.

JTD Keywords: actin, behavior, cell migration, contact guidance, cytoskeleton, fibroblasts, focal adhesions, matrix, microtubules, stiffness, stress fibers, topography, transduction, Contact guidance, Substrate stiffness, Topography

The lack of predictive models that mimic the blood-brain barrier (BBB) hinders the development of effective drugs for neurodegenerative diseases. Animal models behave differently from humans, are expensive and have ethical constraints. Organ-on-a-chip (OoC) platforms offer several advantages to resembling physiological and pathological conditions in a versatile, reproducible, and animal-free manner. In addition, OoC give us the possibility to incorporate sensors to determine cell culture features such as trans-endothelial electrical resistance (TEER). Here, we developed a BBB-on-a-chip (BBB-oC) platform with a TEER measurement system in close distance to the barrier used for the first time for the evaluation of the permeability performance of targeted gold nanorods for theranostics of Alzheimer's disease. GNR-PEG-Ang2/D1 is a therapeutic nanosystem previously developed by us consisting of gold nanorods (GNR) functionalized with polyethylene glycol (PEG), angiopep-2 peptide (Ang2) to overcome the BBB and the D1 peptide as beta amyloid fibrillation inhibitor, finally obtaining GNR-PEG-Ang2/D1 which showed to be useful for disaggregation of the amyloid in in vitro and in vivo models. In this work, we evaluated its cytotoxicity, permeability, and some indications of its impact on the brain endothelium by employing an animal-free device based on neurovascular human cells.In this work, we fabricated a BBB-oC with human astrocytes, pericytes and endothelial cells and a TEER measuring system (TEER-BBB-oC) integrated at a micrometric distance of the endothelial barrier. The characterization displayed a neurovascular network and the expression of tight junctions in the endothelium. We produced GNR-PEG-Ang2/D1 and determined its non-cytotoxic range (0.05-0.4 nM) for plated cells included in the BBB-oC and confirmed its harmless effect at the highest concentration (0.4 nM) in the microfluidic device. The permeability assays revealed that GNR-PEG-Ang2/D1 cross the BBB and this entry is facilitated by Ang2 peptide. Parallel to the permeability analysis of GNR-PEG-Ang2/D1, an interesting behavior of the TJs expression was observed after its administration probably related to the ligands on the nanoparticle surface.BBB-oC with a novel TEER integrated setup which allow a correct read-out and cell imaging monitoring was proven as a functional and throughput platform to evaluate the brain permeability performance of nanotherapeutics in a physiological environment with human cells, putting forward a viable alternative to animal experimentation.© 2023. The Author(s).

JTD Keywords: alzheimer disease (ad), cell-culture, cytotoxicity, endothelial-cells, gold nanoparticles, microfluidic platform, model, organ-on-a-chip (ooc), peptide, tight junction, trans-endothelial electrical resistance (teer), transport, Alzheimer disease (ad), Blood-brain barrier (bbb), Blood-brain-barrier, Blood–brain barrier (bbb), Gold nanoparticles, Organ-on-a-chip (ooc), Trans-endothelial electrical resistance (teer)

Direct lineage reprogramming of one somatic cell into another without transitioning through a progenitor stage has emerged as a strategy to generate clinically relevant cell types. One cell type of interest is the pancreatic insulin-producing β cell whose loss and/or dysfunction leads to diabetes. To date it has been possible to create β-like cells from related endodermal cell types by forcing the expression of developmental transcription factors, but not from more distant cell lineages like fibroblasts. In light of the therapeutic benefits of choosing an accessible cell type as the cell of origin, in this study we set out to analyze the feasibility of transforming human skin fibroblasts into β-like cells. We describe how the timed-introduction of five developmental transcription factors (Neurog3, Pdx1, MafA, Pax4, and Nkx2-2) promotes conversion of fibroblasts toward a β-cell fate. Reprogrammed cells exhibit β-cell features including β-cell gene expression and glucose-responsive intracellular calcium mobilization. Moreover, reprogrammed cells display glucose-induced insulin secretion in vitro and in vivo. This work provides proof-of-concept of the capacity to make insulin-producing cells from human fibroblasts via transcription factor-mediated direct reprogramming.© 2023. The Author(s).

JTD Keywords: adult, beta-cells, differentiation, direct conversion, genes, in-vivo, islets, maturation, pancreatic progenitors, Pluripotent stem-cells

The 3D printing of titanium (Ti) offers countless possibilities for the development of personalized implants with suitable mechanical properties for different medical applications. However, the poor bioactivity of Ti is still a challenge that needs to be addressed to promote scaffold osseointegration. The aim of the present study was to functionalize Ti scaffolds with genetically modified elastin-like recombinamers (ELRs), synthetic polymeric proteins containing the elastin epitopes responsible for their mechanical properties and for promoting mesenchymal stem cell (MSC) recruitment, proliferation, and differentiation to ultimately increase scaffold osseointegration. To this end, ELRs containing specific cell-adhesive (RGD) and/or osteoinductive (SNA15) moieties were covalently attached to Ti scaffolds. Cell adhesion, proliferation, and colonization were enhanced on those scaffolds functionalized with RGD-ELR, while differentiation was promoted on those with SNA15-ELR. The combination of both RGD and SNA15 into the same ELR stimulated cell adhesion, proliferation, and differentiation, although at lower levels than those for every single moiety. These results suggest that biofunctionalization with SNA15-ELRs could modulate the cellular response to improve the osseointegration of Ti implants. Further investigation on the amount and distribution of RGD and SNA15 moieties in ELRs could improve cell adhesion, proliferation, and differentiation compared to the present study.

JTD Keywords: 3d printing, adhesion, biofunctionalization, elastin-like recombinamers, functionalization, hydroxyapatite, osseointegration, polymers, purification, technology, titanium, 3d printing, Surfaces, Titanium

Isotactic polypropylene (i-PP) nonabsorbable surgical meshes are modified by incorporating a conducting polymer (CP) layer to detect the adhesion and growth of bacteria by sensing the oxidation of nicotinamide adenine dinucleotide (NADH), a metabolite produced by the respiration reactions of such microorganisms, to NAD+. A three-step process is used for such incorporation: (1) treat pristine meshes with low-pressure O2 plasma; (2) functionalize the surface with CP nanoparticles; and (3) coat with a homogeneous layer of electropolymerized CP using the nanoparticles introduced in (2) as polymerization nuclei. The modified meshes are stable and easy to handle and also show good electrochemical response. The detection by cyclic voltammetry of NADH within the interval of concentrations reported for bacterial cultures is demonstrated for the two modified meshes. Furthermore, Staphylococcus aureus and both biofilm-positive (B+) and biofilm-negative (B-) Escherichia coli cultures are used to prove real-time monitoring of NADH coming from aerobic respiration reactions. The proposed strategy, which offers a simple and innovative process for incorporating a sensor for the electrochemical detection of bacteria metabolism to currently existing surgical meshes, holds considerable promise for the future development of a new generation of smart biomedical devices to fight against post-operative bacterial infections.

JTD Keywords: adhesion, bacteria metabolism, behavior, biocompatibility, conducting polymer, electrochemical sensor, hernia repair, in-vivo, liquid, nadh detection, plasma treatment, prevention, reinforcement, sensor, smart meshes, Bacteria metabolism, Bacterial infections, Conducting polymer, Electrochemical sensor, Humans, Nad, Nadh detection, Nanoparticles, Oxidation-reduction, Plasma treatment, Polymers, Polypropylene mesh, Smart meshes, Surgical mesh

Lately, there has been an increasing demand for materials that could improve tissue regenerative therapies and provide antimicrobial effects. Similarly, there is a growing need to develop or modify biomaterials for the diagnosis and treatment of different pathologies. In this scenario, hydroxyapatite (HAp) appears as a bioceramic with extended functionalities. Nevertheless, there are certain disadvantages related to the mechanical properties and lack of antimicrobial capacity. To circumvent them, the doping of HAp with a variety of cationic ions is emerging as a good alterative due to the different biological roles of each ion. Among many elements, lanthanides are understudied despite their great potential in the biomedical field. For this reason, the present review focuses on the biological benefits of lanthanides and how their incorporation into HAp can alter its morphology and physical properties. A comprehensive section of the applications of lanthanides-substituted HAp nanoparticles (HAp NPs) is presented to unveil the potential biomedical uses of these systems. Finally, the need to study the tolerable and non-toxic percentages of substitution with these elements is highlighted.

JTD Keywords: biolabeling, biomedicine, biosensors, bone regeneration, calcium, cancer treatment, cationic ions, cell imaging, cerium, doped hap, hydroxyapatite, implants, in-vitro bioactivity, lanthanides-substitutions, lanthanidessubstitutions, nanoparticles, radiation synovectomy, sm-153 particulate hydroxyapatite, structural-characterization, theragnostics, theranostic nanoplatforms, Europium-doped hydroxyapatite, Hydroxyapatite, Theragnostics

The directed migration of cellular clusters enables morphogenesis, wound healing and collective cancer invasion. Gradients of substrate stiffness direct the migration of cellular clusters in a process called collective durotaxis, but the underlying mechanisms remain unclear. Here we unveil a connection between collective durotaxis and the wetting properties of cellular clusters. We show that clusters of cancer cells dewet soft substrates and wet stiff ones. At intermediate stiffness-at the crossover from low to high wettability-clusters on uniform-stiffness substrates become maximally motile, and clusters on stiffness gradients exhibit optimal durotaxis. Durotactic velocity increases with cluster size, stiffness gradient and actomyosin activity. We demonstrate this behaviour on substrates coated with the cell-cell adhesion protein E-cadherin and then establish its generality on substrates coated with extracellular matrix. We develop an active wetting model that explains collective durotaxis in terms of a balance between in-plane active traction and tissue contractility and out-of-plane surface tension. Finally, we show that the distribution of cluster displacements has a heavy tail, with infrequent but large cellular hops that contribute to durotactic migration. Our study demonstrates a physical mechanism of collective durotaxis, through both cell-cell and cell-substrate adhesion ligands, based on the wetting properties of active droplets.

JTD Keywords: Adhesion, Dynamics, E-cadherin, Gradient, Migration, Model, Motility, Movements, Rigidity, Substrate stiffness

Drug and gene delivery systems based on polymeric nanoparticles offer a greater efficacy and a reduced toxicity compared to traditional formulations. Recent studies have evidenced that their internalization, biodistribution and efficacy can be affected, among other factors, by their mechanical properties. Here, we analyze by means of Atomic Force Microscopy force spectroscopy how composition, surface functionalization and loading affect the mechanics of nanoparticles. For this purpose, nanoparticles made of Poly(lactic-co-glycolic) (PLGA) and Ethyl cellulose (EC) with different functionalizations and loading were prepared by nano-emulsion templating using the Phase Inversion Composition method (PIC) to form the nano-emulsions. A multiparametric nanomechanical study involving the determination of the Young's modulus, maximum deformation and breakthrough force was carried out. The obtained results showed that composition, surface functionalization and loading affect the nanomechanical properties in a different way, thus requiring, in general, to consider the overall mechanical properties after the addition of a functionalization or loading. A graphical representation method has been proposed enabling to easily identify mechanically equivalent formulations, which is expected to be useful in the development of soft polymeric nanoparticles for pre-clinical and clinical use.Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.

JTD Keywords: afm, atomic-force microscopy, cell, delivery-systems, drug-delivery, emulsification approach, internalization, mechanics of nanoparticles, nanomedicine, nanoparticle functionalization, particles, protein corona, size, young?s modulus, Afm, Loaded plga nanoparticles, Mechanics of nanoparticles, Nanomedicine, Nanoparticle functionalization, Polymeric nanoparticles, Young’s modulus

Malaria is an infectious disease transmitted by mosquitos, whose control is hampered by drug resistance evolution in the causing agent, protist parasites of the genus Plasmodium, as well as by the resistance of the mosquito to insecticides. New approaches to fight this disease are, therefore, needed. Research into targeted drug delivery is expanding as this strategy increases treatment efficacies. Alternatively, targeting the parasite in humans, here we use single-chain polymer nanoparticles (SCNPs) to target the parasite at the ookinete stage, which is one of the stages in the mosquito. This nanocarrier system provides uniquely sized and monodispersed particles of 5-20 nm, via thiol-Michael addition. The conjugation of succinic anhydride to the SCNP surface provides negative surface charges that have been shown to increase the targeting ability of SCNPs to Plasmodium berghei ookinetes. The biodistribution of SCNPs in mosquitos was studied, showing the presence of SCNPs in mosquito midguts. The presented results demonstrate the potential of anionic SCNPs for the targeting of malaria parasites in mosquitos and may lead to progress in the fight against malaria.

JTD Keywords: antimalarial, atovaquone, carriers, delivery, drug-conjugate, heparin, intramolecular crosslinking, plasmodium berghei, therapy, thiol-michael addition, transmission, Atovaquone, Drug-conjugate, Intramolecular crosslinking, Plasmodium berghei, Plasmodium-falciparum, Single chain polymer nanoparticles, Thiol-michael addition

Mechanical force modulates the conformation and function of individual proteins, and this underpins many mechanically driven cellular processes. This Review addresses single-molecule force spectroscopy experiments conducted on proteins with a known role in mechanosensing and mechanotransduction in eukaryotic cells.; In addition to biochemical signals and genetic considerations, mechanical forces are rapidly emerging as a master regulator of human physiology. However, the molecular mechanisms that regulate force-induced functionalities across a wide range of scales, encompassing the cell, tissue or organ levels, are not well understood in comparison. With the advent, development and refining of single-molecule nanomechanical techniques that enable the conformational dynamics of individual proteins under the effect of a calibrated force to be probed, we have begun to acquire a comprehensive knowledge of the diverse physicochemical principles that regulate the elasticity of single proteins. Here, we review the major advances underpinning our current understanding of how the elasticity of single proteins regulates mechanosensing and mechanotransduction. We discuss the present limitations and future challenges of this prolific and burgeoning field.

JTD Keywords: Cadherin adhesion, Energy landscape, Extracellular-matrix protein, Focal adhesion kinase, Mechanical stability, Molecule force spectroscopy, Muscle protein, N2b element, Stranded-dna, Structural basis

Bioresorbable stents (BRS) are designed to provide initial sufficient mechanical support to prevent vessel recoil while being degraded until their complete resorption. Therefore, degradation rate of BRS plays a crucial role in successful stent performance. This work presents a complete study on the degradation of poly-llactic acid (PLLA) and poly(lactic-co-epsilon-caprolactone) (PLCL) stents fabricated by solvent-cast direct-writing (SC-DW) through two different accelerated assays: alkaline medium at 37 degrees C for 10 days and PBS at 50 degrees C for 4 months. On retrieval, degraded stents were characterized in terms of mass loss, molecular weight (Mw), thermal and mechanical properties. The results showed that under alkaline conditions, stents underwent surface erosion, whereas stents immersed in PBS at 50 degrees C experienced bulk degradation. M-n decrease was accurately described by the autocatalyzed kinetic model, with PLCL showing a degradation rate 1.5 times higher than PLLA. Additionally, stents were subjected to gamma-irradiation and ethylene oxide (EtO) sterilization. Whereas EtOsterilized stents remained structurally unaltered, gamma-irradiated stents presented severe deterioration as a result of extensive chain scission.

JTD Keywords: Acid, Behavior, Bioresorbable stents, Copolymer, Hydrolytic degradation, In-vitro degradation, Mechanical-properties, Molecular-weight, Poly(l-lactide), Poly-l-lactic acid, Poly-l-lactide, Scaffolds, Solvent-cast direct-writing, Sterilization

Heterozygous hTau mice were used for the study of tau seeding. These mice express the six human tau isoforms, with a high predominance of 3Rtau over 4Rtau. The following groups were assessed: (i) non-inoculated mice aged 9 months (n = 4); (ii) Alzheimer's Disease (AD)-inoculated mice (n = 4); (iii) Globular Glial Tauopathy (GGT)-inoculated mice (n = 4); (iv) Pick's disease (PiD)-inoculated mice (n = 4); (v) control-inoculated mice (n = 4); and (vi) inoculated with vehicle alone (n = 2). AD-inoculated mice showed AT8-immunoreactive neuronal pre-tangles, granular aggregates, and dots in the CA1 region of the hippocampus, dentate gyrus (DG), and hilus, and threads and dots in the ipsilateral corpus callosum. GGT-inoculated mice showed unique or multiple AT8-immunoreactive globular deposits in neurons, occasionally extended to the proximal dendrites. PiD-inoculated mice showed a few loose pre-tangles in the CA1 region, DG, and cerebral cortex near the injection site. Coiled bodies were formed in the corpus callosum in AD-inoculated mice, but GGT-inoculated mice lacked globular glial inclusions. Tau deposits in inoculated mice co-localized active kinases p38-P and SAPK/JNK-P, thus suggesting active phosphorylation of the host tau. Tau deposits were absent in hTau mice inoculated with control homogenates and vehicle alone. Deposits in AD-inoculated hTau mice contained 3Rtau and 4Rtau; those in GGT-inoculated mice were mainly stained with anti-4Rtau antibodies, but a small number of deposits contained 3Rtau. Deposits in PiD-inoculated mice were stained with anti-3Rtau antibodies, but rare neuronal, thread-like, and dot-like deposits showed 4Rtau immunoreactivity. These findings show that tau strains produce different patterns of active neuronal seeding, which also depend on the host tau. Unexpected 3Rtau and 4Rtau deposits after inoculation of homogenates from 4R and 3R tauopathies, respectively, suggests the regulation of exon 10 splicing of the host tau during the process of seeding, thus modulating the plasticity of the cytoskeleton.

JTD Keywords: alzheimer's disease (ad), alzheimers-disease, brain, corticobasal degeneration, globular glial tauopathy (ggt), htau, isoforms, pathological tau, pick's disease (pid), picks-disease, propagation, protein, seeding, tau splicing, tauopathy, Alzheimer disease, Alzheimer’s disease (ad), Animals, Brain, Globular glial tauopathy (ggt), Hippocampus, Htau, Humans, Mice, Mice, transgenic, Paired helical filaments, Pick disease of the brain, Pick’s disease (pid), Seeding, Tau, Tau proteins, Tau splicing, Tauopathies

Tissue engineering is nowadays a powerful tool to restore damaged tissues and recover their normal functionality. Advantages over other current methods are well established, although a continuous evolution is still necessary to improve the final performance and the range of applications. Trends are nowadays focused on the development of multifunctional scaffolds with hierarchical structures and the capability to render a sustained delivery of bioactive molecules under an appropriate stimulus. Nanocomposites incorporating hydroxyapatite nanoparticles (HAp NPs) have a predominant role in bone tissue regeneration due to their high capacity to enhance osteoinduction, osteoconduction, and osteointegration, as well as their encapsulation efficiency and protection capability of bioactive agents. Selection of appropriated polymeric matrices is fundamental and consequently great efforts have been invested to increase the range of properties of available materials through copolymerization, blending, or combining structures constituted by different materials. Scaffolds can be obtained from different processes that differ in characteristics, such as texture or porosity. Probably, electrospinning has the greater relevance, since the obtained nanofiber membranes have a great similarity with the extracellular matrix and, in addition, they can easily incorporate functional and bioactive compounds. Coaxial and emulsion electrospinning processes appear ideal to generate complex systems able to incorporate highly different agents. The present review is mainly focused on the recent works performed with Hap-loaded scaffolds having at least one structural layer composed of core/shell nanofibers.

JTD Keywords: bone tissue, coaxial electrospinning, composite nanofibers, drug-release behavior, emulsion electrospinning, hydroxyapatite, in-vitro evaluation, mechanical-properties, osteogenic differentiation, pickering emulsions, protein adsorption, structured scaffolds, surface-initiated polymerization, tissue regeneration, Bone tissue, Coaxial electrospinning, Emulsion electrospinning, Hydroxyapatite, Multifunctional scaffolds, Poly(3-hydroxybutyrate) phb patches, Tissue regeneration

Directed cell migration is an essential building block of life, present when an embryo develops, a dendritic cell migrates toward a lymphatic vessel, or a fibrotic organ fails to restore its normal parenchyma. Directed cell migration is often guided by spatial gradients in a physicochemical property of the cell microenvironment, such as a gradient in chemical factors dissolved in the medium or a gradient in the mechanical properties of the substrate. Single cells and tissues sense these gradients, establish a back-to-front polarity, and coordinate the migration machinery accordingly. Central to these steps we find physical forces. In some cases, these forces are integrated into the gradient sensing mechanism. Other times, they transmit information through cells and tissues to coordinate a collective response. At any time, they participate in the cellular migratory system. In this review, we explore the role of physical forces in gradient sensing, polarization, and coordinating movement from single cells to multicellular collectives. We use the framework proposed by the molecular clutch model and explore to what extent asymmetries in the different elements of the clutch can lead to directional migration.

JTD Keywords: Cell migration, Cell traction forces, Chemotaxis, Contact guidance, Curvotaxis, Cytoskeleton, Durotaxis, Extracellular-matrix, Focal adhesions, Guidance, Haptotaxis, Leading-edge, Mechanotaxis, Membrane tension, Molecular clutch model, Myosin-ii, Reduced inequalities, Rigidity, Stress fibers, Traction

JTD Keywords: biocompatibility, blends, design, dopamine detection, electrocatalytic oxidation, electrodes, polymers, sensor, Additive manufacturing, Surface

Around 30-40% of patients with colorectal cancer (CRC) undergoing curative resection of the primary tumour will develop metastases in the subsequent years1. Therapies to prevent disease relapse remain an unmet medical need. Here we uncover the identity and features of the residual tumour cells responsible for CRC relapse. An analysis of single-cell transcriptomes of samples from patients with CRC revealed that the majority of genes associated with a poor prognosis are expressed by a unique tumour cell population that we named high-relapse cells (HRCs). We established a human-like mouse model of microsatellite-stable CRC that undergoes metastatic relapse after surgical resection of the primary tumour. Residual HRCs occult in mouse livers after primary CRC surgery gave rise to multiple cell types over time, including LGR5+ stem-like tumour cells2-4, and caused overt metastatic disease. Using Emp1 (encoding epithelial membrane protein 1) as a marker gene for HRCs, we tracked and selectively eliminated this cell population. Genetic ablation of EMP1high cells prevented metastatic recurrence and mice remained disease-free after surgery. We also found that HRC-rich micrometastases were infiltrated with T cells, yet became progressively immune-excluded during outgrowth. Treatment with neoadjuvant immunotherapy eliminated residual metastatic cells and prevented mice from relapsing after surgery. Together, our findings reveal the cell-state dynamics of residual disease in CRC and anticipate that therapies targeting HRCs may help to avoid metastatic relapse.© 2022. The Author(s), under exclusive licence to Springer Nature Limited.

JTD Keywords: colonization, defines, human colon, mutations, plasticity, retrieval, stem-cells, subtypes, underlie, Animal, Animal cell, Animal experiment, Animal model, Animal tissue, Animals, Article, Cancer, Cancer growth, Cancer immunotherapy, Cancer inhibition, Cancer recurrence, Cancer staging, Cell, Cell adhesion, Cell migration, Cell population, Cell surface receptor, Cohort analysis, Colorectal cancer, Colorectal neoplasms, Colorectal tumor, Comprehensive molecular characterization, Controlled study, Crispr-cas9 system, Cytoskeleton, Disease exacerbation, Disease progression, Dynamics, Emp1 gene, Epithelial membrane protein-1, Extracellular matrix, Flow cytometry, Fluorescence intensity, Gene expression, Genetics, Human, Human cell, Humans, Immune response, Immunofluorescence, In situ hybridization, Marker gene, Metastasis potential, Mice, Minimal residual disease, Mouse, Neoplasm proteins, Neoplasm recurrence, local, Neoplasm, residual, Nonhuman, Pathology, Phenotype, Prevention and control, Protein, Receptors, cell surface, Single cell rna seq, Tumor, Tumor protein, Tumor recurrence

Mesenchymal condensation is a prevalent morphogenetic transition that is essential in chondrogenesis. However, the current understanding of condensation mechanisms is limited. In vivo, progenitor cells directionally migrate from the surrounding loose mesenchyme towards regions of increasing matrix adherence (the condensation centers), which is accompanied by the upregulation of fibronectin. Here, we focused on the mechanisms of cell migration during mesenchymal cell condensation and the effects of matrix adherence. Dendrimer-based nanopatterns of the cell-adhesive peptide arginine-glycine-aspartic acid (RGD), which is present in fibronectin, were used to regulate substrate adhesion. We recorded collective and single-cell migration of mesenchymal stem cells, under chondrogenic induction, using live-cell imaging. Our results show that the cell migration mode of single cells depends on substrate adhesiveness, and that cell directionality controls cell condensation and the fusion of condensates. Inhibition experiments revealed that cell???cell interactions mediated by N-cadherin (also known as CDH2) are also pivotal for directional migration of cell condensates by maintaining cell???cell cohesion, thus suggesting a fine interplay between cell???matrix and cell???cell adhesions. Our results shed light on the role of cell interactions with a fibronectin-depositing matrix during chondrogenesis in vitro, with possible applications in regenerative medicine.

JTD Keywords: Alpha-v-beta-3, Arginine-glycine-aspartic acid, Cell migration, Chondrogenesis, Dynamics, Expression, Fibronectin, Gastrulation, Involvement, Mechanisms, Mesenchymal condensation, Model, N-cadherin, Nanopatterned substrates, Rgd

JTD Keywords: anti-fxiia antibody, artificial surfaces, blood compatibility, complement activation, factor xii, fibrinolytic system, hemocompatible coatings, interactive hemocompatibility, poly(2-methacryloyloxyethyl phosphorylcholine), polyethylene oxide, polymer brushes, radical polymerization, sequential coimmobilization, Antifouling polymer brushes, Protein adsorption

Whenever an artificial surface comes into contact with blood, proteins are rapidly adsorbed onto its surface. This phenomenon, termed fouling, is then followed by a series of undesired reactions involving activation of complement or the coagulation cascade and adhesion of leukocytes and platelets leading to thrombus formation. Thus, considerable efforts are directed towards the preparation of fouling-resistant surfaces with the best possible hemocompatibility. Herein, a comprehensive hemocompatibility study after heparinized blood contact with seven polymer brushes prepared by surface-initiated atom transfer radical polymerization is reported. The resistance to fouling is quantified and thrombus formation and deposition of blood cellular components on the coatings are analyzed. Moreover, identification of the remaining adsorbed proteins is performed via mass spectroscopy to elucidate their influence on the surface hemocompatibility. Compared with an unmodified glass surface, the grafting of polymer brushes minimizes the adhesion of platelets and leukocytes and prevents the thrombus formation. The fouling from undiluted blood plasma is reduced by up to 99%. Most of the identified proteins are connected with the initial events of foreign body reaction towards biomaterial (coagulation cascade proteins, complement component, and inflammatory proteins). In addition, several proteins that are not previously linked with blood-biomaterial interaction are presented and discussed.

JTD Keywords: antifouling surfaces, biosensor, blood-plasma, coagulation, coatings, compatibility, glycoprotein, hemocompatibility, identification, methacrylate), ms identification, polymer brushes, protein adsorption, surface-chemistry, Antifouling surfaces, High-density-lipoprotein, Protein adsorption

Hydroxyapatite nanoparticles are popular tools in bone regeneration, but they have also been used for gene delivery and as anticancer drugs. Understanding their mechanism of action, particularly for the latter application, is crucial to predict their toxicity. To this end, we aimed to elucidate the importance of nanoparticle membrane interactions in the cytotoxicity of MG-63 cells using two different types of nanoparticles. In addition, conventional techniques for studying nanoparticle internalisation were evaluated and compared with newer and less exploited approaches. Hydroxyapatite and magnesium-doped hydroxyapatite nanoparticles were used as suspensions or compacted as specular discs. Comparison between cells seeded on the discs and those supplemented with the nanoparticles allowed direct interaction of the cell membrane with the material to be ruled out as the main mechanism of toxicity. In addition, standard techniques such as flow cytometry were inconclusive when used to assess nanoparticles toxicity. Interestingly, the use of intracellular calcium fluorescent probes revealed the presence of a high number of calcium-rich vesicles after nanoparticle supplementation in cell culture. These structures could not be detected by transmission electron microscopy due to their liquid content. However, by using cryo-soft X-ray imaging, which was used to visualise the cellular ultrastructure without further treatment other than vitrification and to quantify the linear absorption coefficient of each organelle, it was possible to identify them as multivesicular bodies, potentially acting as calcium stores. In the study, an advanced state of degradation of the hydroxyapatite and magnesium-doped hydroxyapatite nanoparticles within MG-63 cells was observed. Overall, we demonstrate that the combination of fluorescent calcium probes together with cryo-SXT is an excellent approach to investigate intracellular calcium, especially when found in its soluble form.Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.

JTD Keywords: adsorption, cryo-soft x-ray tomography, cytotoxicity, expression, flow cytometry, internalisation, intracellular calcium, magnesium, nano, nanomaterials, nanoparticles, proliferation, protein corona, ultrastructure, Calcium-phosphate nanoparticles, Cryo-soft x-ray tomography, Flow cytometry, Hydroxyapatite, Internalisation, Intracellular calcium, Nanoparticles

Several factors present in the extracellular environment regulate epithelial cell adhesion and dynamics. Among them, growth factors such as EGF, upon binding to their receptors at the cell surface, get internalized and directly activate the acto-myosin machinery. In this study we present the effects of EGF on the contractility of epithelial cancer cell colonies in confined geometry of different sizes. We show that the extent to which EGF triggers contractility scales with the cluster size and thus the number of cells. Moreover, the collective contractility results in a radial distribution of traction forces, which are dependent on integrin β1 peripheral adhesions and transmitted to neighboring cells through adherens junctions. Taken together, EGF-induced contractility acts on the mechanical crosstalk and linkage between the cell-cell and cell-matrix compartments, regulating collective responses.Copyright © 2022 The Authors. Published by Elsevier GmbH.. All rights reserved.

JTD Keywords: actin, activation, actomyosin, adherens junctions, adhesion, e-cadherin, egf, maturation, mechanical regulation, micropatterning, migration, traction forces, transduction, transmission, Actomyosin, Adherens junctions, Cell adhesion, Cell membrane, Collective contractility, Egf, Epidermal growth factor, Epidermal-growth-factor, Epithelial cells, Micropatterning, Myosins, Traction forces

Local cancer treatment by in situ injections of thermo-responsive hydrogels (HG) offers several advantages over conventional systemic anti-cancer treatments. In this work, a biodegradable and multicompartmental HG composed of N-isopropylacrylamide, cellulose, citric acid, and ceric ammonium nitrate was developed for the controlled release of hydrophilic (doxorubicin) and hydrophobic (niclosamide) drugs. The formulation presented ideal properties regarding thermo-responsiveness, rheological behavior, drug release profile, biocompatibility, and biological activity in colon and ovarian cancer cells. Cellulose was found to retard drugs release rate, being only 4 % of doxorubicin and 30 % of niclosamide released after 1 week. This low release was sufficient to cause cell death in both cell lines. Moreover, HG demonstrated a proper injectability, in situ prevalence, and safety profile in vivo. Overall, the HG properties, together with its natural and eco-friendly composition, create a safe and efficient platform for the local treatment of non-resectable tumors or tumors requiring pre-surgical adjuvant therapy.

JTD Keywords: biodegradable, cellulose, controlled-release formulation, drug delivery systems, hydrogel, thermo-responsiveness, Ammonium-nitrate, Biodegradable, Cancer treatment, Cellulose, Controlled-release formulation, Delivery, Drug delivery systems, Hydrogel, Reduce, Thermo-responsiveness

Calcium phosphate cements (CPCs) are attractive synthetic bone grafts as they possess osteoconductive and osteoinductive properties. Their biomimetic synthesis grants them an intrinsic nano-and microporosity that resembles natural bone and is paramount for biological processes such as protein adhesion, which can later enhance cell adhesion. However, a main limitation of CPCs is the lack of macroporosity, which is crucial to allow cell colonization throughout the scaffold. Moreover, CPCs lack specific motifs to guide cell interactions through their membrane proteins. In this study, we explore a strategy targeting simultaneously both macroporosity and cell binding motifs within CPCs by the use of recombinant silk. A silk protein functionalized with the cell binding motif RGD serves as foaming template of CPCs to achieve biomimetic hydroxyapatite (HA) scaffolds with multiscale porosity. The synergies of RGD-motifs in the silk macroporous template and the biomimetic features of HA are explored for their potential to enhance mesenchymal stem cell adhesion, proliferation, migration and differentiation. Macroporous Silk-HA scaffolds improve initial cell adhesion compared to a macroporous HA in the absence of silk, and importantly, the presence of silk greatly enhances cell migration into the scaffold. Additionally, cell proliferation and osteogenic differentiation are achieved in the scaffolds.

JTD Keywords: Bioceramics, Bone, Bone regeneration, Composites, Degradation, Fabrication, Hydroxyapatite, Hydroxyapatite scaffolds, Injectability, Porosity, Recombinant spider silk, Rgd motifs, Silk, Stem-cells

JTD Keywords: adhesion, attachment, growth, ligands, membrane, microcontact printing, migration, organoid-derived intestinal epithelia, receptor, tissue organization, Eph-ephrin, Stem-cells

Gradients of signaling pathways within the intestinal stem cell (ISC) niche are instrumental for cellular compartmentalization and tissue function, yet how are they sensed by the epithelium is still not fully understood. Here a new in vitro model of the small intestine based on primary epithelial cells (i), apically accessible (ii), with native tissue mechanical properties and controlled mesh size (iii), 3D villus-like architecture (iv), and precisely controlled biomolecular gradients of the ISC niche (v) is presented. Biochemical gradients are formed through hydrogel-based scaffolds by free diffusion from a source to a sink chamber. To confirm the establishment of spatiotemporally controlled gradients, light-sheet fluorescence microscopy and in-silico modeling are employed. The ISC niche biochemical gradients coming from the stroma and applied along the villus axis lead to the in vivo-like compartmentalization of the proliferative and differentiated cells, while changing the composition and concentration of the biochemical factors affects the cellular organization along the villus axis. This novel 3D in vitro intestinal model derived from organoids recapitulates both the villus-like architecture and the gradients of ISC biochemical factors, thus opening the possibility to study in vitro the nature of such gradients and the resulting cellular response.© 2022 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.

JTD Keywords: 3d architectures, biomolecular gradients, colon, crypt, engineering organoids, hydrogels, identification, in silico modeling, intestinal stem cell niches, light sheet fluorescence microscopy, niche, permeability, photolithography, regeneration, villus, wnt, 3d architectures, Biomolecular gradients, Engineering organoids, In silico modeling, Intestinal stem cell niches, Light sheet fluorescence microscopy, Photolithography, Stem-cell

We report a green solvent-to-polymer upgrading transformation of chemicals of the lactic acid portfolio into water-soluble lower critical solution temperature (LCST)-type acrylic polymers. Aqueous Cu(0)-mediated living radical polymerization (SET-LRP) was utilized for the rapid synthesis of N-substituted lactamide-type homo and random acrylic copolymers under mild conditions. A particularly unique aspect of this work is that the water-soluble monomers and the SET-LRP initiator used to produce the corresponding polymers were synthesized from biorenewable and non-toxic solvents, namely natural ethyl lactate and BASF's Agnique (R) AMD 3L (N,N-dimethyl lactamide, DML). The pre-disproportionation of Cu(I) Br in the presence of tris[2-(dimethylamino)ethyl]amine (Me6TREN) in water generated nascent Cu(0) and Cu(II) complexes that facilitated the fast polymerization of N-tetrahydrofurfuryl lactamide and N,N-dimethyl lactamide acrylate monomers (THFLA and DMLA, respectively) up to near-quantitative conversion with excellent control over molecular weight (5000 < M-n < 83 000) and dispersity (1.05 < D < 1.16). Interestingly, poly(THFLA) showed a degree of polymerization and concentration dependent LCST behavior, which can be fine-tuned (T-cp = 12-62 degrees C) through random copolymerization with the more hydrophilic DMLA monomer. Finally, covalent cross-linking of these polymers resulted in a new family of thermo-responsive hydrogels with excellent biocompatibility and tunable swelling and LCST transition. These illustrate the versatility of these neoteric green polymers in the preparation of smart and biocompatible soft materials.

JTD Keywords: Acid, Ethyl lactate, Living radical polymerization, Monomers, Pnipam, Reductive amination, Ruthenium nanoparticles, Set-lrp, Single, Thermoresponsive polymers

Identification of asymptomatic patients at higher risk for suffering cardiac events remains controversial and challenging in Brugada syndrome (BS). In this work, we proposed an ECG-based classifier to predict BS-related symptoms, by merging the most predictive electrophysiological features derived from the ventricular depolarization and repolarization periods, along with autonomic-related markers. The initial feature space included local and dynamic ECG markers, assessed during a physical exercise test performed in 110 BS patients (25 symptomatic). Morphological, temporal and spatial properties quantifying the ECG dynamic response to exercise and recovery were considered. Our model was obtained by proposing a two-stage feature selection process that combined a resampled-based regularization approach with a wrapper model assessment for balancing, simplicity and performance. For the classification step, an ensemble was constructed by several logistic regression base classifiers, whose outputs were fused using a performance-based weighted average. The most relevant predictors corresponded to the repolarization interval, followed by two autonomic markers and two other makers of depolarization dynamics. Our classifier allowed for the identification of novel symptom-related markers from autonomic and dynamic ECG responses during exercise testing, suggesting the need for multifactorial risk stratification approaches in order to predict future cardiac events in asymptomatic BS patients.

JTD Keywords: brugada syndrome, depolarization disorders, ensemble classifier, heart-rate recovery, Acute myocardial-ischemia, Autonomic nervous system, Autonomics, Brugada syndrome, Brugadum syndrome, Cardiac death, Death, sudden, cardiac, Depolarization, Depolarization disorder, Depolarization disorders, Dynamic ecg, Electrocardiography, Electrophysiology, Ensemble classifier, Ensemble-classifier, Events, Exercise, Exercise test, Forecasting, Heart, Heart rate, Heart-rate, Heart-rate recovery, Humans, Prognosis, Qrs, Quantification, Recovery, Repolarization, Sudden cardiac death

Cells are subjected to multiple mechanical inputs throughout their lives. Their ability to detect these environmental cues is called mechanosensing, a process in which integrins play an important role. During cellular mechanosensing, plasma membrane (PM) tension is adjusted to mechanical stress through the buffering action of caveolae; however, little is known about the role of caveolae in early integrin mechanosensing regulation. Here, we show that Cav1KO fibroblasts increase adhesion to FN-coated beads when pulled with magnetic tweezers, as compared to wild type fibroblasts. This phenotype is Rho-independent and mainly derived from increased active b1-integrin content on the surface of Cav1KO fibroblasts. FRAP analysis and endocytosis/recycling assays revealed that active b1-integrin is mostly endocytosed through the CLIC/GEEC pathway and is more rapidly recycled to the PM in Cav1KO fibroblasts, in a Rab4 and PM tension-dependent manner. Moreover, the threshold for PM tension-driven b1-integrin activation is lower in Cav1KO MEFs than in wild type MEFs, through a mechanism dependent on talin activity. Our findings suggest that caveolae couple mechanical stress to integrin cycling and activation, thereby regulating the early steps of the cellular mechanosensing response.© 2022, Lolo et al.

JTD Keywords: adhesion, alpha-v-beta-3, cell, integrin activation, internalization, kinase, mechanosensing, mediated endocytosis, mouse, stiffness, talin, trafficking, Animals, Caveolae, Cell adhesion, Cell biology, Fibroblasts, Integrin activation, Integrin beta1, Integrin recycling, Integrins, Mechanosensing, Membrane tension, Mice, Mouse, Stress, mechanical

The synthesis and study of the tripeptide Arg-Gly-Asp (RGD), the binding site of different extracellular matrix proteins, e.g., fibronectin and vitronectin, has allowed the production of a wide range of cell adhesive surfaces. Although the surface density and spacing of the RGD peptide at the nanoscale have already shown a significant influence on cell adhesion, the impact of its hierarchical nanostructure is still rather unexplored. Accordingly, a versatile colloidal system named quatsomes, based on fluid nanovesicles formed by the self-assembling of cholesterol and surfactant molecules, has been devised as a novel template to achieve hierarchical nanostructures of the RGD peptide. To this end, RGD was anchored on the vesicle's fluid membrane of quatsomes, and the RGD-functionalized nanovesicles were covalently anchored to planar gold surfaces, forming a state of quasi-suspension, through a long poly(ethylene glycol) (PEG) chain with a thiol termination. An underlying self-assembled monolayer (SAM) of a shorter PEG was introduced for vesicle stabilization and to avoid unspecific cell adhesion. In comparison with substrates featuring a homogeneous distribution of RGD peptides, the resulting hierarchical nanoarchitectonic dramatically enhanced cell adhesion, despite lower overall RGD molecules on the surface. The new versatile platform was thoroughly characterized using a multitechnique approach, proving its enhanced performance. These findings open new methods for the hierarchical immobilization of biomolecules on surfaces using quatsomes as a robust and novel tissue engineering strategy.

JTD Keywords: activation, arg-gly-asp (rgd), cell adhesion, extracellular-matrix, growth, integrins, ligands, nanopatterns, quatsomes, scaffolds, self-assembled monolayers, surface engineering, tissue engineering, Arg-gly-asp (rgd), Cell adhesion, Integrins, Nano-structured surfaces, Nanovesicles, Quatsomes, Self-assembled monolayers, Surface engineering, Tissue engineering

Polarized hydroxyapatite (p-HAp), a calcium phosphate catalyst obtained at high temperature under intense electric field, has been used for the synthesis of ammonium starting from N2 and liquid water at low pressure (<6 bar) and temperatures below 120 C. The success of the nitrogen fixation process has been demonstrated by isotope labelling experiments using 15N2. Considering the optimal reaction conditions for the production of ammonium, the yield is as high as 154.6 +/- 25.8 mu mol/g of catalyst. The proposed synthesis exhibits three important advantages for its utilization in green chemistry environmental processes related to the recycling of polluted air. These are: i) the catalysts converts CO2 into valuable chemical products in addition of transforming N2 in ammonium; ii) the final energy balance is very favorable since no external electrical field is necessary to promote nitrogen and carbon fixation reactions; and iii) products are easily transferred to water favoring their extraction and avoiding the saturation of the catalyst.

JTD Keywords: Adsorbed nitrogen, Air pollution, Amino-acids, Electrophotosynthesis, Environmental process, Facile synthesis, Fixation, Functionalization, Hydroxyapatite, Nitride, Nitrogen reduction, Polarized catalyst

Epithelial cell divisions are coordinated with cell loss to preserve epithelial integrity. However, how epithelia adapt their rate of cell division to changes in cell number, for instance during homeostatic turnover or wounding, is not well understood. Here, we show that epithelial cells sense local cell density through mechanosensitive E-cadherin adhesions to control G2/M cell-cycle progression. As local cell density increases, tensile forces on E-cadherin adhesions are reduced, which prompts the accumulation of the G2 checkpoint kinase Wee1 and downstream inhibitory phosphorylation of Cdk1. Consequently, dense epithelia contain a pool of cells that are temporarily halted in G2 phase. These cells are readily triggered to divide following epithelial wounding due to the consequent increase in intercellular forces and resulting degradation of Wee1. Our data collectively show that epithelial cell division is controlled by a mechanical G2 checkpoint, which is regulated by cell-density-dependent intercellular forces sensed and transduced by E-cadherin adhesions.Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.

JTD Keywords: Adherens junction, Cadherins, Cell cycle, Cell cycle proteins, Cell division, Cp: cell biology, E-cadherin, Epithelial cells, Epithelial homeostasis, G2 checkpoint, G2 phase cell cycle checkpoints, Mechanical forces, Mechanotransduction, Mitosis, Phosphorylation, Proliferation

Cooperative behaviour of sperm is one of the mechanisms that plays a role in sperm competition. It has been observed in several species that spermatozoa interact with each other to form agglomerates or bundles. In this study, we investigate the effect of physical and biochemical factors that will most likely promote bundle formation in bull sperm. These factors include fluid viscosity, swim-up process, post-thaw incubation time and media additives which promote capacitation. While viscosity does not seem to influence the degree of sperm bundling, swim-up, post-thaw migration time and suppressed capacitation increase the occurrence of sperm bundles. This leads to the conclusion that sperm bundling is a result of hydrodynamic and adhesive interactions between the cells which occurs frequently during prolonged incubation times.Copyright © 2022 Morcillo i Soler, Hidalgo, Fekete, Zalanyi, Khalil, Yeste and Magdanz.

JTD Keywords: acrosome reaction, adhesion, bundling, capacitation, cell-cell interaction, cooperation, cooperative behaviour, fertilization, mammals, membrane, motility, progesterone, sperm competition, sperm migration, sperm selection, Bovine spermatozoa, Bundling, Cell-cell interaction, Cooperative behaviour, Sperm competition, Sperm migration, Sperm selection, Spermatozoa

Upon the initiation of collective cell migration, the cells at the free edge are specified as leader cells; however, the mechanism underlying the leader cell specification remains elusive. Here, we show that lamellipodial extension after the release from mechanical confinement causes sustained extracellular signal-regulated kinase (ERK) activation and underlies the leader cell specification. Live-imaging of Madin-Darby canine kidney (MDCK) cells and mouse epidermis through the use of Förster resonance energy transfer (FRET)-based biosensors showed that leader cells exhibit sustained ERK activation in a hepatocyte growth factor (HGF)-dependent manner. Meanwhile, follower cells exhibit oscillatory ERK activation waves in an epidermal growth factor (EGF) signaling-dependent manner. Lamellipodial extension at the free edge increases the cellular sensitivity to HGF. The HGF-dependent ERK activation, in turn, promotes lamellipodial extension, thereby forming a positive feedback loop between cell extension and ERK activation and specifying the cells at the free edge as the leader cells. Our findings show that the integration of physical and biochemical cues underlies the leader cell specification during collective cell migration.Copyright © 2022 Elsevier Inc. All rights reserved.

JTD Keywords: activation, c-met, contact inhibition, focal adhesions, heparan-sulfate, mechanical forces, morphogenesis, rho, stress fibers, Collective cell migration, Erk, Feedback regulation, Fret, Growth-factor receptor, Hgf, Lamellipodia, Leader cell specification, Signal transduction, Traction force, Wound healing

JTD Keywords: biochemical engineering, neurotrauma and neurodegenerative disease, optogenetics and dreadds, spinal cord injury (sci), Biochemical engineering, Neurotrauma and neurodegenerative disease, Optogenetics and dreadds, Sensorimotor, Spinal cord injury (sci)

Cannabis is one of the most commonly used substances in the world. However, its effects on human cognition are not yet fully understood. Although the cerebellum has the highest density of cannabinoid receptor type 1 (CB1R) in the human brain, literature on how cannabis use affects cerebellar-dependent learning is sparse. This study examined the effect of chronic cannabis use on sensorimotor adaptation, a cerebellar-mediated task, which has been suggested to depend on endocannabinoid signaling.Chronic cannabis users (n = 27) with no psychiatric comorbidities and healthy, cannabis-naïve controls (n = 25) were evaluated using a visuomotor rotation task. Cannabis users were re-tested after 1 month of abstinence (n = 13) to assess whether initial differences in performance would persist after cessation of use.Cannabis users showed lower adaptation rates compared to controls at the first time point. However, this difference in performance did not persist when participants were retested after one month of abstinence (n = 13). Healthy controls showed attenuated implicit learning in the late phase of the adaptation during re-exposure, which was not present in cannabis users. This explains the lack of between group differences in the second time point and suggests a potential alteration of synaptic plasticity required for cerebellar learning in cannabis users.Overall, our results suggest that chronic cannabis users show alterations in sensorimotor adaptation, likely due to a saturation of the endocannabinoid system after chronic cannabis use.Copyright © 2022 Elsevier Ltd. All rights reserved.

JTD Keywords: Addiction, Cannabis, Cerebellum, Endocannabinoid system, Visuomotor adaptation

Tauopathies are a group of neurodegenerative diseases characterized by the hyperphosphorylation and deposition of tau proteins in the brain. In Alzheimer's disease, and other related tauopathies, the pattern of tau deposition follows a stereotypical progression between anatomically connected brain regions. Increasing evidence suggests that tau behaves in a "prion-like" manner, and that seeding and spreading of pathological tau drive progressive neurodegeneration. Although several advances have been made in recent years, the exact cellular and molecular mechanisms involved remain largely unknown. Since there are no effective therapies for any tauopathy, there is a growing need for reliable experimental models that would provide us with better knowledge and understanding of their etiology and identify novel molecular targets. In this review, we will summarize the development of cellular models for modeling tau pathology. We will discuss their different applications and contributions to our current understanding of the "prion-like" nature of pathological tau.

JTD Keywords: neurodegeneration, seeding, spreading, Culture model, Efficient generation, Extracellular tau, Familial alzheimers-disease, Microtubule-associated protein, Mouse model, Neurodegeneration, Neurofibrillary tangles, Paired helical filaments, Pathogenic tau, Pluripotent stem-cells, Seeding, Spreading, Tauopathies

Great advances in cancer treatment have been undertaken in the last years as a consequence of the development of new antitumoral drugs able to target cancer cells with decreasing side effects and a better understanding of the behavior of neoplastic cells during invasion and metastasis. Specifically, drug delivery systems (DDS) based on the use of hydroxyapatite nanoparticles (HAp NPs) are gaining attention and merit a comprehensive review focused on their potential applications. These are derived from the intrinsic properties of HAp (e.g., biocompatibility and biodegradability), together with the easy functionalization and easy control of porosity, crystallinity and morphology of HAp NPs. The capacity to tailor the properties of DLS based on HAp NPs has well-recognized advantages for the control of both drug loading and release. Furthermore, the functionalization of NPs allows a targeted uptake in tumoral cells while their rapid elimination by the reticuloendothelial system (RES) can be avoided. Advances in HAp NPs involve not only their use as drug nanocarriers but also their employment as nanosystems for magnetic hyperthermia therapy, gene delivery systems, adjuvants for cancer immunotherapy and nanoparticles for cell imaging.

JTD Keywords: antitumoral, cancer, cell imaging, controlled-release, drug-carrier, efficient drug-delivery, fatty-acid-metabolism, fe3o4 nanoparticles, gene delivery, hydroxyapatite, hyperthermia, immunotherapy, in-vitro, magnetic hydroxyapatite, nano-hydroxyapatite, protein adsorption, tumor-growth, Calcium-phosphate nanoparticles, Cancer, Immunotherapy

Cell-material interactions are regulated by mimicking bone extracellular matrix on the surface of biomaterials. In this regard, reproducing the extracellular conditions that promote integrin and growth factor (GF) signaling is a major goal to trigger bone regeneration. Thus, the use of synthetic osteogenic domains derived from bone morphogenetic protein 2 (BMP-2) is gaining increasing attention, as this strategy is devoid of the clinical risks associated with this molecule. In this work, the wrist and knuckle epitopes of BMP-2 are screened to identify peptides with potential osteogenic properties. The most active sequences (the DWIVA motif and its cyclic version) are combined with the cell adhesive RGD peptide (linear and cyclic variants), to produce tailor-made biomimetic peptides presenting the bioactive cues in a chemically and geometrically defined manner. Such multifunctional peptides are next used to functionalize titanium surfaces. Biological characterization with mesenchymal stem cells demonstrates the ability of the biointerfaces to synergistically enhance cell adhesion and osteogenic differentiation. Furthermore, in vivo studies in rat calvarial defects prove the capacity of the biomimetic coatings to improve new bone formation and reduce fibrous tissue thickness. These results highlight the potential of mimicking integrin-GF signaling with synthetic peptides, without the need for exogenous GFs.© 2022 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.

JTD Keywords: adhesion formation, bmp-2, cell adhesions, in-vivo, integrin, mesenchymal stem-cells, morphogenetic protein-2, multifunctionality, osteoblastic differentiation, osteogenic differentiation, rgd-dwiva, rgd-peptides, titanium biofunctionalization, titanium surfaces, Animals, Biocompatible materials, Biomimetic peptides, Bone morphogenetic protein 2, Bone regeneration, Cell adhesions, Cell differentiation, Epitopes, Extracellular matrix, Integrins, Marrow stromal cells, Multifunctionality, Osteogenesis, Osteogenic differentiation, Peptides, Rats, Rgd-dwiva, Titanium, Titanium biofunctionalization

Most of the conventional in vitro models to test biomaterial-driven vascularization are too simplistic to recapitulate the complex interactions taking place in the actual cell microenvironment, which results in a poor prediction of the in vivo performance of the material. However, during the last decade, cell culture models based on microfluidic technology have allowed attaining unprecedented levels of tissue biomimicry. In this work, we propose a microfluidic-based 3D model to evaluate the effect of bioactive biomaterials capable of releasing signalling cues (such as ions or proteins) in the recruitment of endogenous endothelial progenitor cells, a key step in the vascularization process. The usability of the platform is demonstrated using experimentally-validated finite element models and migration and proliferation studies with rat endothelial progenitor cells (rEPCs) and bone marrow-derived rat mesenchymal stromal cells (BM-rMSCs). As a proof of concept of biomaterial evaluation, the response of rEPCs to an electrospun composite made of polylactic acid with calcium phosphates nanoparticles (PLA+CaP) was compared in a co-culture microenvironment with BM-rMSC to a regular PLA control. Our results show a significantly higher rEPCs migration and the upregulation of several pro-inflammatory and proangiogenic proteins in the case of the PLA+CaP. The effects of osteopontin (OPN) on the rEPCs migratory response were also studied using this platform, suggesting its important role in mediating their recruitment to a calcium-rich microenvironment. This new tool could be applied to screen the capacity of a variety of bioactive scaffolds to induce vascularization and accelerate the preclinical testing of biomaterials. STATEMENT OF SIGNIFICANCE: : For many years researchers have used neovascularization models to evaluate bioactive biomaterials both in vitro, with low predictive results due to their poor biomimicry and minimal control over cell cues such as spatiotemporal biomolecule signaling, and in vivo models, presenting drawbacks such as being highly costly, time-consuming, poor human extrapolation, and ethically controversial. We describe a compact microphysiological platform designed for the evaluation of proangiogenesis in biomaterials through the quantification of the level of sprouting in a mimicked endothelium able to react to gradients of biomaterial-released signals in a fibrin-based extracellular matrix. This model is a useful tool to perform preclinical trustworthy studies in tissue regeneration and to better understand the different elements involved in the complex process of vascularization.Copyright © 2022. Published by Elsevier Ltd.

JTD Keywords: angiogenesis, bioactive materials, bone regeneration, bone-formation, calcium-phosphate, extracellular calcium, in-vitro, interstitial flow, ion release, microfluidic model, signalling gradient, substitutes, tissue engineering, vascularization, vegf, Ion release, Mesenchymal stem-cells, Tissue engineering, Vascularization

Electrochemical sensors for real-time detection of several bioanalytes have been prepared by additive manufacturing, shaping non-conductive poly(lactic acid) (PLA) filaments, and applying a physical treatment to create excited species. The latter process, which consists of the application of power discharge of 100 W during 2 min in a chamber at a low pressure of O-2, converts electrochemically inert PLA into an electrochemically responsive material. The electric discharge caused the oxidation of the PLA surface as evidenced by the increment in the quantity of oxygenated species detected by FTIR spectroscopy and X-ray photoelectron spectroscopy (XPS). Indeed, changes in the surface chemical composition became more pronounced with increasing O-2 pressure. After demonstrating the performance of the chemically modified material as individual dopamine and glucose sensors, multiplexed detection has been achieved by measuring simultaneously the two voltammetric signals. This has been performed by collecting the signals in two different regions, a naked chemically modified PLA for dopamine detection and a chemically modified PLA region functionalized with Glucose Oxidase. These outcomes led to define a new paradigm for manufacturing electrodes for electrochemical sensors based on 3D printing without using conducting materials at any stage of the process.

JTD Keywords: Additive manu f a c turing, Carbon, Conductivity, Degradation, Dopamine, Dopamine detection, Glucose detection, Glucose sensors, Immobilization, Multiplexed detect i o n, Oxidase, Plasma treatment

Nanosized artificial antigen-presenting cells (aAPCs), synthetic immune cell mimics that aim to activate T cells ex or in vivo, offer an effective alternative to cellular immunotherapies. However, comprehensive studies that delineate the effect of nano-aAPC topology, including nanoparticle morphology and ligand density, are lacking. Here, we systematically studied the topological effects of polymersome-based aAPCs on T cell activation. We employed an aAPC library created from biodegradable poly(ethylene glycol)-block-poly(d,l-lactide) (PEG-PDLLA) polymersomes with spherical or tubular shape and variable sizes, which were functionalized with αCD3 and αCD28 antibodies at controlled densities. Our results indicate that high ligand density leads to enhancement in T cell activation, which can be further augmented by employing polymersomes with larger size. At low ligand density, the effect of both polymersome shape and size was more pronounced, showing that large elongated polymersomes better activate T cells compared to their spherical or smaller counterparts. This study demonstrates the capacity of polymersomes as aAPCs and highlights the role of topology for their rational design.

JTD Keywords: antibody density, artificial antigen-presenting cells, biodegradable polymersomes, design, expansion, immunotherapy, nano-immunotherapy, nanoparticle morphology, t cell activation, Biodegradable polymersomes, Nanoparticle morphology, Synthetic dendritic cells

Aims: Cognitive impairment in patients with alcohol use disorder (AUD) is highly prevalent, and it negatively impacts treatment outcome. However, this condition is neither systematically assessed nor treated. Thus, we aimed to explore the usability of a virtual reality-based protocol ('Rehabilitation Gaming System', RGS) for patients with AUD. Methods: Twenty AUD patients (50% also cognitive impairment) underwent a single session of the RGS protocol (four cognitive training tasks, 10 minutes each). System Usability Scale (SUS) and Post-Study System Usability Questionnaire (PSSUQ) were applied to assess the RGS usability and patients' satisfaction with it. Also, the Perceived Competence Scale was administered to assess the patients' feelings of competence when using the training protocol. Comparisons of the responses to these questionnaires were performed between AUD patients with cognitive impairment and those without cognitive impairment. Results: RGS usability was very positively rated (median SUS score = 80, Interquartile Range, IQR = 68.13-86-88). No significant differences were found in the median SUS scores for any of the sociodemographic or clinical variables, excepting for gender (women median score = 85; IQR = 80-94.38 vs. men median score = 71.25; IQR = 61.25-89.25; P-value = 0.035). The quality of the information provided by the RGS training scenarios and the usability were positively rated (PSSUQ), and patients experienced high feelings of competence. Conclusions: The RGS has been found to be usable in the short term and patients with AUD stated to be satisfied with it. Future larger, randomized trials are needed to explore the effectiveness of this tool to help overcome the cognitive deficits in AUD patients. Short Summary: Although cognitive impairments are highly prevalent in alcohol use disorder (AUD), no long-term gold standard intervention has yet been identified. The Rehabilitation Gaming System (virtual reality-based cognitive training protocol) has shown short-term high usability in AUD. Its effectiveness in providing engaging, long-term cognitive rehabilitation in AUD should be further assessed.

JTD Keywords: Addiction, Brain-damage, Deficits, Impact, Neurocognitive impairment, Therapy

Although alginate is widely used as a matrix in the formulation of cell-laden inks, this polymer often requires laborious processing strategies due to its lack of cell adhesion moieties. The main objective of the present work was to explore the incorporation of microspheres into alginate-based bioinks as a simple and tuneable way to solve the cell adhesion problems, while adding extra biological functionality and improving their mechanical properties. To this end, three types of microspheres with different mineral contents (i.e. gelatine with 0% of hydroxyapatite, gelatine with 25 wt% of hydroxyapatite nanoparticles and 100 wt% of calcium -deficient hydroxyapatite) were synthesised and incorporated into the formulation of cell-laden inks. The results showed that the addition of microspheres generally improved the rheological properties of the ink, favoured cell proliferation and positively affected osteogenic cell differentiation. Furthermore, this differentiation was found to be influenced by the type of microsphere and the ability of the cells to migrate towards them, which was highly dependent on the stiffness of the bioink. In this regard, Ca2+ supplementation in the cell culture medium had a pronounced effect on the relaxation of the stiffness of these cell-loaded inks, influencing the overall cell performance. In conclusion, we have developed a powerful and tuneable strategy for the fabrication of alginate-based bioinks with enhanced biological characteristics by incorporating microspheres into the initial ink formulation.; [GRAPHICS]; .

JTD Keywords: 3d bioprinting, alginate, bioink, gelatine, hydroxyapatite, 3d bioprinting, Alginate, Behavior, Bioink, Cell-culture, Gelatin, Gelatine, Hydrogels, Hydroxyapatite, Laden, Microspheres, Mineralization, Scaffolds

According to circular economy, wine-making by-products represent a fascinating biomass, which can be used for the sustainable exploitation of polyphenols and the development of new nanotechnological health-promoting products. In this study, polyphenols contained in the grape pomace were extracted by maceration with ethanol in an easy and low dissipative way. The obtained extract, rich in malvidin-3-glucoside, quercetin, pro-cyanidin B2 and gallic acid, was incorporated into phospholipid vesicles tailored for intestinal delivery. To improve their performances, vesicles were enriched with gelatine or a maltodextrin (Nutriose (R)), or their com-bination (gelatine-liposomes, nutriosomes and gelatine-nutriosomes). The small (-147 nm) and negatively charged (--50mV) vesicles were stable at different pH values mimicking saliva (6.75), gastric (1.20) and intestinal (7.00) environments. Vesicles effectively protected intestinal cells (Caco-2) from the oxidative stress and promoted the biofilm formation by probiotic bacteria. A preliminary evaluation of the vesicle feasibility at industrial levels was also performed, analysing the economic and energetic costs needed for their production.

JTD Keywords: Adhesion, Antioxidant activity, Caco-2 cells, Dextrin, Grape pomace extract, Lactobacillus-reuteri, Manufacturing costs, Oxidative stress, Ph, Phospholipid vesicles, Polyphenols, Probiotic bacteria, Protein

Tissue inhibitor of metalloproteinase-1 (TIMP-1) is an important regulator of extracellular matrix turnover that has been traditionally regarded as a potential tumor suppressor owing to its inhibitory effects of matrix metal-loproteinases. Intriguingly, this interpretation has been challenged by the consistent observation that increased expression of TIMP-1 is associated with poor prognosis in virtually all cancer types including lung cancer, supporting a tumor-promoting function. However, how TIMP-1 is dysregulated within the tumor micro-environment and how it drives tumor progression in lung cancer is poorly understood. We analyzed the expression of TIMP-1 and its cell surface receptor CD63 in two major lung cancer subtypes: lung adenocarci-noma (ADC) and squamous cell carcinoma (SCC), and defined the tumor-promoting effects of their interac-tion. We found that TIMP-1 is aberrantly overexpressed in tumor-associated fibroblasts (TAFs) in ADC compared to SCC. Mechanistically, TIMP-1 overexpression was mediated by the selective hyperactivity of the pro-fibrotic TGF-61/SMAD3 pathway in ADC-TAFs. Likewise, CD63 was upregulated in ADC compared to SCC cells. Genetic analyses revealed that TIMP-1 secreted by TGF-61-activated ADC-TAFs is both nec-essary and sufficient to enhance growth and invasion of ADC cancer cells in culture, and that tumor cell expression of CD63 was required for these effects. Consistently, in vivo analyses revealed that ADC cells co-injected with fibroblasts with reduced SMAD3 or TIMP-1 expression into immunocompromised mice attenu-ated tumor aggressiveness compared to tumors bearing parental fibroblasts. We also found that high TIMP1 and CD63 mRNA levels combined define a stronger prognostic biomarker than TIMP1 alone. Our results identify an excessive stromal TIMP-1 within the tumor microenvironment selectively in lung ADC, and implicate it in a novel tumor-promoting TAF-carcinoma crosstalk, thereby pointing to TIMP-1/CD63 interaction as a novel therapeutic target in lung cancer. (c) 2022 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)

JTD Keywords: cancer-associated fibroblast, cd63, fibrosis, smad3, tgf-β1, timp-1, Angiogenesis, Cancer cells, Cancer-associated fibroblast, Cd63, Expression, Fibrosis, Hepatocellular-carcinoma, Metalloproteinases, Nintedanib, Prognostic-significance, Protein, Smad3, Squamous-cell carcinoma, Tgf-? 1, Tgf-β1, Timp-1, Tissue inhibitor, Tumor microenvironment

Gene duplications occur in prokaryotic genomes at a detectable frequency. In many instances, the biological function of the duplicates is unknown, and hence, the significance of the presence of multiple copies of these genes remains unclear.; Gene duplications significantly impact the gene repertoires of both eukaryotic and prokaryotic microorganisms. The genomes of pathogenic Escherichia coli strains share a group of duplicated genes whose function is mostly unknown. The irmA gene is one of the duplicates encoded in several pathogenic E. coli strains. The function of its gene product was investigated in the uropathogenic E. coli strain CFT073, which contains a single functional copy. The IrmA protein structure mimics that of human interleukin receptors and likely plays a role during infection. The enteroaggregative E. coli strain 042 contains two functional copies of the irmA gene. In the present work, we investigated their biological roles. The irmA_4509 allele is expressed under several growth conditions. Its expression is modulated by the global regulators OxyR and Hha, with optimal expression at 37 degrees C and under nutritional stress conditions. Expression of the irmA_2244 allele can only be detected when the irmA_4509 allele is knocked out. Differences in the promoter regions of both alleles account for their differential expression. Our results show that under several environmental conditions, the expression of the IrmA protein in strain 042 is dictated by the irmA_4509 allele. The irmA_2244 allele appears to play a backup role to ensure IrmA expression when the irmA_4509 allele loses its function. IMPORTANCE Gene duplications occur in prokaryotic genomes at a detectable frequency. In many instances, the biological function of the duplicates is unknown, and hence, the significance of the presence of multiple copies of these genes remains unclear. In pathogenic E. coli isolates, the irmA gene can be present either as a single copy or in two or more copies. We focused our work on studying why a different pathogenic E. coli strain encodes two functional copies of the irmA gene. We show that under several environmental conditions, one of the alleles dictates IrmA expression, and the second remains silent. The latter allele is only expressed when the former is silenced. The presence of more than one functional copy of the irmA gene in some pathogenic E. coli strains can result in sufficient expression of this virulence factor during the infection process.

JTD Keywords: 042, aec69, enteroaggregative e. coli, gene duplications, 042, Adaptation, Aec69, Aggregative adherence, Alleles, Chromosomal genes, Coli, Duplication, Enteroaggregative e, Enteroaggregative e. coli, Escherichia coli infections, Escherichia coli proteins, Escherichia-coli, Evolution, Gene duplications, Hha/ymoa, Humans, Irma, Mechanism, Outer-membrane, Protein, Uropathogenic escherichia coli

Chronic use of cannabis leads to both motor deficits and the downregulation of CB1 receptors (CB1R) in the cerebellum. In turn, cerebellar damage is often related to impairments in motor learning and control. Further, a recent motor learning task that measures cerebellar-dependent adaptation has been shown to distinguish well between healthy subjects and chronic cannabis users. Thus, the deteriorating effects of chronic cannabis use in motor performance point to cerebellar adaptation as a key process to explain such deficits. We review the literature relating chronic cannabis use, the endocannabinoid system in the cerebellum, and different forms of cerebellar-dependent motor learning, to suggest that CB1R downregulation leads to a generalized underestimation and misprocessing of the sensory errors driving synaptic updates in the cerebellar cortex. Further, we test our hypothesis with a computational model performing a motor adaptation task and reproduce the behavioral effect of decreased implicit adaptation that appears to be a sign of chronic cannabis use. Finally, we discuss the potential of our hypothesis to explain similar phenomena related to motor impairments following chronic alcohol dependency. © 2022

JTD Keywords: adaptation, addiction, alcohol-abuse, cerebellum, chronic cannabis use, cognition, deficits, endocannabinoid system, error processing, explicit, modulation, motor learning, release, synaptic plasticity, Adaptation, Adaptation, physiological, Alcoholism, Article, Behavioral science, Cannabinoid 1 receptor, Cannabis, Cannabis addiction, Cerebellum, Cerebellum cortex, Cerebellum disease, Chronic cannabis use, Computer model, Down regulation, Endocannabinoid, Endocannabinoid system, Endocannabinoids, Error processing, Hallucinogens, Human, Humans, Motor dysfunction, Motor learning, Nerve cell plasticity, Nonhuman, Physiology, Psychedelic agent, Purkinje-cells, Regulatory mechanism, Sensation, Sensory dysfunction, Sensory error processing impairment, Synaptic transmission, Task performance

The combination of microfluidics and photo-polymerization techniques such as stereolithography (SLA) has emerged as a new field which has a lot of potential to influence in such important areas as biological analysis, and chemical detection among others. However, the integration between them is still at an early stage of development. In this article, after analyzing the resolution of a custom SLA 3D printer with commercial resins, microfluidic devices were manufactured using three different approaches. First, printing a mold with the objective of creating a Polydimethylsiloxane (PDMS) replica with the microfluidic channels; secondly, open channels have been printed and then assembled with a flat cover of the same resin material. Finally, a closed microfluidic device has also been produced in a single process of printing. Important results for 3D printing with commercial resins have been achieved by only printing one layer on top of the channel. All microfluidic devices have been tested successfully for pressure-driven fluid flow.

JTD Keywords: 3d printing, additive manufacturing, microfluidics, photo-curable polymers, 3d printing, Additive manufacturing, Microfluidics, Photo-curable polymers, Stereolithography

Type 1 Diabetes results from autoimmune response elicited against β-cell antigens. Nowadays, insulin injections remain the leading therapeutic option. However, injection treatment fails to emulate the highly dynamic insulin release that β-cells provide. 3D cell-laden microspheres have been proposed during the last years as a major platform for bioengineering insulin-secreting constructs for tissue graft implantation and a model for in vitro drug screening platforms. Current microsphere fabrication technologies have several drawbacks: the need for an oil phase containing surfactants, diameter inconsistency of the microspheres, and high time-consuming processes. These technologies have widely used alginate for its rapid gelation, high processability, and low cost. However, its low biocompatible properties do not provide effective cell attachment. This study proposes a high-throughput methodology using a 3D bioprinter that employs an ECM-like microenvironment for effective cell-laden microsphere production to overcome these limitations. Crosslinking the resulting microspheres with tannic acid prevents collagenase degradation and enhances spherical structural consistency while allowing the diffusion of nutrients and oxygen. The approach allows customization of microsphere diameter with extremely low variability. In conclusion, a novel bio-printing procedure is developed to fabricate large amounts of reproducible microspheres capable of secreting insulin in response to extracellular glucose stimuli.© 2022 The Authors. Advanced Materials Technologies published by Wiley‐VCH GmbH.

JTD Keywords: 3d bioprinter, beta-cell, biomaterial, collagen, encapsulation, mechanics, microspheres, survival, 3d bioprinter, ?-cell, Advanced material technologies, Biocompatibility, Cell encapsulations, Cells, Collagen, Cross-linking, Cytology, Drug delivery, Encapsulation, Fabrication, Flavonoids, Gelation, In-vitro, Insulin injections, Insulin release, Microspheres, Tannic acid, Tannins, Throughput, Tissue grafts, Type 1 diabetes, Β‐cell

Moderate exercise has well-founded benefits in cardiovascular health. However, increasing, yet controversial, evidence suggests that extremely trained athletes may not be protected from cardiovascular events as much as moderately trained individuals. In our rodent model, intensive but not moderate training promoted aorta and carotid stiffening and elastic lamina ruptures, tunica media thickening of intramyocardial arteries, and an imbalance between vasoconstrictor and relaxation agents. An up-regulation of angiotensin-converter enzyme, miR-212, miR-132, and miR-146b might account for this deleterious remodeling. Most changes remained after a 4-week detraining. In conclusion, our results suggest that intensive training blunts the benefits of moderate exercise. (C) 2022 The Authors. Published by Elsevier on behalf of the American College of Cardiology Foundation.

JTD Keywords: atherosclerosis, cacs, coronary artery calcium score, cad, coronary artery disease, coronary artery disease, cv, cardiovascular, endurance exercise, extreme sport, mmp9, matrix metalloproteinase 9, no, nitric oxide, phe, phenylephrine, vsmc, vascular smooth muscle cell, Age, Atherosclerosis, Cacs, coronary artery calcium score, Cad, coronary artery disease, Coronary artery disease, Coronary atherosclerosis, Cv, cardiovascular, Disease, Endurance exercise, Extreme sport, Metalloproteinases, Micrornas, Mmp9, matrix metalloproteinase 9, No, nitric oxide, Phe, phenylephrine, Physical-activity, Prevalence, Rats, Relevance, Risk, Vascular stiffening, Vsmc, vascular smooth muscle cell

Over the last decades, photopharmacology has gone far beyond its proof-of-concept stage to become a bona fide approach to study neural systems in vivo. Indeed, photopharmacological control has expanded over a wide range of endogenous targets, such as receptors, ion channels, transporters, kinases, lipids, and DNA transcription processes. In this review, we provide an overview of the recent progresses in the in vivo photopharmacological control of neuronal circuits and behavior. In particular, the use of small aquatic animals for the in vivo screening of photopharmacological compounds, the recent advances in optical modulation of complex behaviors in mice, and the development of adjacent techniques for light and drug delivery in vivo are described.

JTD Keywords: brain circuits, circadian rhythm, in vivo photomodulation, in vivo technology, neuronal receptors, Architecture, Azobenzene photoswitches, Brain circuits, Channels, Circadian rhythm, In vivo photomodulation, In vivo technology, Light, Modulator, Neuronal receptors, Optical control, Optogenetics, Pharmacology, Photopharmacology, Receptors, Systems

We investigated the diffusion of three cyclic boronates formulated as beta-lactamase inhibitors through the porin OmpF to evaluate their potential to cross OM via the porin pathway. The three nonbeta-lactam molecules diffuse through the porin eyelet region with the same mechanism observed for beta-lactam molecules and diazobicyclooctan derivatives, with the electric dipole moment aligned with the transversal electric field. In particular, the BOH group can interact with both the basic ladder and the acidic loop L3, which is characteristic of the size-constricted region of this class of porins. On one hand, we confirm that the transport of small molecules through enterobacter porins has a common general mechanism; on the other, the class of cyclic boronate molecules does not seem to have particular difficulties in diffusing through enterobacter porins, thus representing a good scaffold for new anti-infectives targeting Gram-negative bacteria research.

JTD Keywords: beta-lactamase inhibitors, cyclic boronates, diffusion current, metadynamics, molecular dynamics simulations, permeation, Antibiotics, Beta-lactamase inhibitors, Cyclic boronates, Diffusion, Diffusion current, Discovery, Electric-field, Metadynamics, Molecular dynamics simulations, Molecular-dynamics simulations, Nanopores, Permeability, Permeation, Porins, Rules, Translocation

This study explored the use of parasternal second intercostal space and lower intercostal space surface electromyogram (sEMG) and surface mechanomyogram (sMMG) recordings (sEMGpara and sMMGpara, and sEMGlic and sMMGlic, respectively) to assess neural respiratory drive (NRD), neuromechanical (NMC) and neuroventilatory (NVC) coupling, and mechanical efficiency (MEff) noninvasively in healthy subjects and chronic obstructive pulmonary disease (COPD) patients. sEMGpara, sMMGpara, sEMGlic, sMMGlic, mouth pressure (Pmo), and volume (Vi) were measured at rest, and during an inspiratory loading protocol, in 16 COPD patients (8 moderate and 8 severe) and 9 healthy subjects. Myographic signals were analyzed using fixed sample entropy and normalized to their largest values (fSEsEMGpara%max, fSEsMMGpara%max, fSEsEMGlic%max, and fSEsMMGlic%max). fSEsMMGpara%max, fSEsEMGpara%max, and fSEsEMGlic%max were significantly higher in COPD than in healthy participants at rest. Parasternal intercostal muscle NMC was significantly higher in healthy than in COPD participants at rest, but not during threshold loading. Pmo-derived NMC and MEff ratios were lower in severe patients than in mild patients or healthy subjects during threshold loading, but differences were not consistently significant. During resting breathing and threshold loading, Vi-derived NVC and MEff ratios were significantly lower in severe patients than in mild patients or healthy subjects. sMMG is a potential noninvasive alternative to sEMG for assessing NRD in COPD. The ratios of Pmo and Vi to sMMG and sEMG measurements provide wholly noninvasive NMC, NVC, and MEff indices that are sensitive to impaired respiratory mechanics in COPD and are therefore of potential value to assess disease severity in clinical practice. Author

JTD Keywords: biomedical measurement, chronic obstructive pulmonary disease, couplings, diaphragm, disease severity, efficiency, electromyography, exacerbations, healthy volunteers, inspiratory muscles, loading, mechanomyography, obstructive pulmonary-disease, pressure measurement, protocols, respiratory mechanics, respiratory muscles, responsiveness, spirometry, stimulation, volume measurement, At rests, Biomedical measurement, Biomedical measurements, Chronic obstructive pulmonary disease, Couplings, Disease severity, Efficiency ratio, Electromyography, Healthy subjects, Healthy volunteers, Loading, Mechanical efficiency, Mechanomyogram, Muscle, Muscles, Neural respiratory drive, Noninvasive medical procedures, Pressure measurement, Protocols, Pulmonary diseases, Surface electromyogram, Volume measurement

Biological adhesion is a critical mechanical function of complex organisms. At the scale of cell-cell contacts, adhesion is remarkably tunable to enable both cohesion and malleability during development, homeostasis and disease. It is physically supported by transient and laterally mobile molecular bonds embedded in fluid membranes. Thus, unlike specific adhesion at solid-solid or solid-fluid interfaces, peeling at fluid-fluid interfaces can proceed by breaking bonds, by moving bonds or by a combination of both. How the additional degree of freedom provided by bond mobility changes the mechanics of peeling is not understood. To address this, we develop a theoretical model coupling diffusion, reactions and mechanics. Mobility and reaction rates determine distinct peeling regimes. In a diffusion-dominated Stefan-like regime, bond motion establishes self-stabilizing dynamics that increase the effective fracture energy. In a reaction-dominated regime, peeling proceeds by travelling fronts where marginal diffusion and unbinding control peeling speed. In a mixed reaction-diffusion regime, strengthening by bond motion competes with weakening by bond breaking in a force-dependent manner, defining the strength of the adhesion patch. In turn, patch strength depends on molecular properties such as bond stiffness, force sensitivity or crowding. We thus establish the physical rules enabling tunable cohesion in cellular tissues and in engineered biomimetic systems.

JTD Keywords: cell–cell adhesion, peeling, Adhesive contact, Cadherins, Cell-cell adhesion, Detachment, Detailed mechanics, Diffusion, Growth, Kinetics, Peeling, Red-blood-cells, Repulsion, Separation, Vesicle adhesion

Biomolecular gradients are widely present in multiple biological processes. Historically they were reproduced in vitro by using micropipettes, Boyden and Zigmond chambers, or hydrogels. Despite the great utility of these setups in the study of gradient-related problems such as chemotaxis, they face limitations when trying to translate more complex in vivo-like scenarios to in vitro systems. In the last 20 years, the advances in manufacturing of micromechanical systems (MEMS) had opened the possibility of applying this technology to biology (BioMEMS). In particular, microfluidics has proven extremely efficient in setting-up biomolecular gradients which are stable, controllable, reproducible and at length scales that are relevant to cells. In this chapter, we give an overview of different methods to generate molecular gradients using microfluidics, then we discuss the different steps of the pipeline to fabricate a gradient generator microfluidic device, and at the end, we show an application example of the fabrication of a microfluidic device that can be used to generate a surface-bound biomolecular gradient.© 2022. The Author(s), under exclusive license to Springer Nature Switzerland AG.

JTD Keywords: biomems, gradient, microfluidics, model, nanotechnology, proteins, Biomems, Gradient, Mechanisms, Microfabrication, Microfluidics, Nanotechnology

JTD Keywords: biosensor, blood-plasma, chemistry, coatings, design, microchannel cantilever spotting, strain-promoted alkyne-azide cycloaddition, ultra-low fouling hierarchical polymer brushes, Alkyne cycloaddition, Coupling efficiency

Working memory has been shown to rely on theta oscillations' phase synchronicity for item encoding and recall. At the same time, saccadic eye movements during visual exploration have been observed to trigger theta-phase resets, raising the question of whether the neuronal substrates of mnemonic processing rely on motor-evoked responses. To quantify the relationship between saccades and working memory load, we recorded eye tracking and behavioral data from human participants simultaneously performing an n-back Sternberg auditory task and a hue-based catch detection task. In addition to task-specific interference in performance, we also found that saccade rate was modulated by working memory load in the Sternberg task's preresponse stage. Our results support the possibility of interplay between saccades and hippocampal theta during working memory retrieval of items.

JTD Keywords: Eeg, Eye-tracking technology, Hippocampus, Humans, Memory, short-term, Mental recall, Microsaccades, Normality, Parietal cortex, Persistent activity, Prefrontal cortex, Saccades

Invasive lobular breast carcinoma (ILC) is characterized by proliferative indolence and long-term latency relapses. This study aimed to identify how disseminating ILC cells control the balance between quiescence and cell cycle re-entry. In the absence of anchorage, ILC cells undergo a sustained cell cycle arrest in G0/G1 while maintaining viability. From the genes that are upregulated in anchorage independent ILC cells, we selected Inhibitor of DNA binding 2 (Id2), a mediator of cell cycle progression. Using loss-of-function experiments, we demonstrate that Id2 is essential for anchorage independent survival (anoikis resistance) in vitro and lung colonization in mice. Importantly, we find that under anchorage independent conditions, E-cadherin loss promotes expression of Id2 in multiple mouse and (organotypic) human models of ILC, an event that is caused by a direct p120-catenin/Kaiso-dependent transcriptional de-repression of the canonical Kaiso binding sequence TCCTGCNA. Conversely, stable inducible restoration of E-cadherin expression in the ILC cell line SUM44PE inhibits Id2 expression and anoikis resistance. We show evidence that Id2 accumulates in the cytosol, where it induces a sustained and CDK4/6-dependent G0/G1 cell cycle arrest through interaction with hypo-phosphorylated Rb. Finally, we find that Id2 is indeed enriched in ILC when compared to other breast cancers, and confirm cytosolic Id2 protein expression in primary ILC samples. In sum, we have linked mutational inactivation of E-cadherin to direct inhibition of cell cycle progression. Our work indicates that loss of E-cadherin and subsequent expression of Id2 drive indolence and dissemination of ILC. As such, E-cadherin and Id2 are promising candidates to stratify low and intermediate grade invasive breast cancers for the use of clinical cell cycle intervention drugs.

JTD Keywords: anoikis resistance, carcinoma, d1, differentiation, gene-expression, growth, id2, proliferation, repression, Mammary epithelial-cells

Since the discovery that nanostructured surfaces were able to kill bacteria, many works have been published focusing on the design of nanopatterned surfaces with antimicrobial properties. Synthetic bone grafts, based on calcium phosphate (CaP) formulations, can greatly benefit from this discovery if adequate nanotopographies can be developed. However, CaP are reactive materials and experience ionic exchanges when placed into aqueous solutions which may in turn affect cell behaviour and complicate the interpretation of the bactericidal results. The present study explores the bactericidal potential of two nanopillared CaP prepared by hydrolysis of two different sizes of alpha-tricalcium phosphate (alpha-TCP) powders under biomimetic or hydrothermal conditions. A more lethal bactericidal response toward Pseudomonas aeruginosa (similar to 75% killing efficiency of adhered bacteria) was obtained from the hydrothermally treated CaP which consisted in a more irregular topography in terms of pillar size (radius: 20-60 nm), interpillar distances (100-1500 nm) and pillar distribution (pillar groups forming bouquets) than the biomimetically treated one (radius: 20-40 nm and interpillar distances: 50-200 nm with a homogeneous pillar distribution). The material reactivity was greatly influenced by the type of medium (nutrient-rich versus nutrient-free) and the presence or not of bacteria. A lower reactivity and superior bacterial attachment were observed in the nutrient-free medium while a lower attachment was observed for the nutrient rich medium which was explained by a superior reactivity of the material paired with the lower tendency of planktonic bacteria to adhere on surfaces in the presence of nutrients. Importantly, the ionic exchanges produced by the presence of materials were not toxic to planktonic cells. Thus, we can conclude that topography was the main contributor to mortality in the bacterial adhesion tests.

JTD Keywords: bactericidal, calcium deficient hydroxyapatite, calcium phosphates, nanopillars, pseudomonas aeruginosa, reactivity, Adhesion, Anti-bacterial agents, Antibacterial, Bacterial adhesion, Bactericidal, Biomaterials, Biomimetics, Calcium deficient hydroxyapatite, Calcium phosphates, Hydroxyapatite, In-vitro, Infections, Nanopillars, Nanostructures, Pseudomonas aeruginosa, Pseudomonas-aeruginosa, Reactivity, Recent progress, Silver, Topography, Transmission

Alzheimer’s disease (AD) and other tauopathies are common neurodegenerative diseases in older adults; in contrast, abnormal tau deposition in neurons and glial cells occurs only exceptionally in children. Sarkosyl-insoluble fractions from sporadic AD (sAD) containing paired helical filaments (PHFs) were inoculated unilaterally into the thalamus in newborn and three-month-old wild-type C57BL/6 mice, which were killed at different intervals from 24 h to six months after inoculation. Tau-positive cells were scanty and practically disappeared at three months in mice inoculated at the age of a newborn. In contrast, large numbers of tau-positive cells, including neurons and oligodendrocytes, were found in the thalamus of mice inoculated at three months and killed at the ages of six months and nine months. Mice inoculated at the age of newborn and re-inoculated at the age of three months showed similar numbers and distribution of positive cells in the thalamus at six months and nine months. This study shows that (a) differences in tau seeding between newborn and young adults may be related to the ratios between 3Rtau and 4Rtau, and the shift to 4Rtau predominance in adults, together with the immaturity of connections in newborn mice, and (b) intracerebral inoculation of sAD PHFs in newborn mice does not protect from tau seeding following intracerebral inoculation of sAD PHFs in young/adult mice.

JTD Keywords: alzheimer's disease, alzheimer-disease, alzheimer’s disease, expression, mouse tau, neurofibrillary tangles, newborn, pathological tau, propagation, protein-tau, spread, tau seeding and spreading, thalamus, transgenic mice, Alzheimer disease, Alzheimer’s disease, Animals, Brain, Mice, Mice, inbred c57bl, Mice, transgenic, Neurofibrillary tangles, Newborn, Paired helical filaments, Tau proteins, Tau seeding and spreading, Tauopathies, Thalamus

One of the main limitations of in vitro studies on lung diseases is the difficulty of maintaining the type II phenotype of alveolar epithelial cells in culture. This fact has previously been related to the translocation of the mechanosensing Yes-associated protein (YAP) to the nuclei and Rho signaling pathway. In this work, we aimed to culture and subculture primary alveolar type II cells on extracellular matrix lung-derived hydrogels to assess their suitability for phenotype maintenance. Cells cultured on lung hydrogels formed monolayers and maintained type II phenotype for a longer time as compared with those conventionally cultured. Interestingly, cells successfully grew when they were subsequently cultured on a dish. Moreover, cells cultured on a plate showed the active form of the YAP protein and the formation of stress fibers and focal adhesions. The results of chemically inhibiting the Rho pathway strongly suggest that this is one of the mechanisms by which the hydrogel promotes type II phenotype maintenance. These results regarding protein expression strongly suggest that the chemical and biophysical properties of the hydrogel have a considerable impact on the transition from ATII to ATI phenotypes. In conclusion, culturing primary alveolar epithelial cells on lung ECM-derived hydrogels may facilitate the prolonged culturing of these cells, and thus help in the research on lung diseases.

JTD Keywords: adhesion, alveolar cells, expression, extracellular matrix, hydrogels, pathway, surfactant, type ii phenotype, yap, Extracellular matrix, Transplantation, Type ii phenotype

Aim: To unveil the influence of cell-matrix adhesions in the establishment of gap junction intercellular communication (GJIC) during cell condensation in chondrogenesis. Materials & methods: Previously developed nanopatterns of the cell adhesive ligand arginine-glycine-aspartic acid were used as cell culture substrates to control cell adhesion at the nanoscale. In vitro chondrogenesis of mesenchymal stem cells was conducted on the nanopatterns. Cohesion and GJIC were evaluated in cell condensates. Results: Mechanical stability and GJIC are enhanced by a nanopattern configuration in which 90% of the surface area presents adhesion sites separated less than 70 nm, thus providing an onset for cell signaling. Conclusion: Cell-matrix adhesions regulate GJIC of mesenchymal cell condensates during in vitro chondrogenesis from a threshold configuration at the nanoscale.

JTD Keywords: arginine-glycine-aspartic acid, arginine–glycine–aspartic acid, cell adhesion, condensation, dendrimer-based nanopatterning, gap junction intercellular communication, Actin, Adhesion, Arginine-glycine-aspartic acid, Cell adhesion, Collagen, Condensation, Connexin-43, Dendrimer-based nanopatterning, Dynamics, Extracellular-matrix, Fibronectin, Gap junction intercellular communication, Mesenchymal stem cells, Permeability, Phenotype, Vinculin

The influence of the properties of different solid substrates on the tethering of two antibodies, IgG1-CR3022 and IgG1-S309, which were specifically engineered for the detection of SARS-CoV-2, has been examined at the molecular level using conventional and accelerated Molecular Dynamics (cMD and aMD, respectively). Two surfaces with very different properties and widely used in immunosensors for diagnosis, amorphous silica and the most stable facet of the face-centered cubic gold structure, have been considered. The effects of such surfaces on the structure and orientation of the immobilized antibodies have been determined by quantifying the tilt and hinge angles that describe the orientation and shape of the antibody, respectively, and the dihedrals that measure the relative position of the antibody arms with respect to the surface. Results show that the interactions with amorphous silica, which are mainly electrostatic due to the charged nature of the surface, help to preserve the orientation and structure of the antibodies, especially of the IgG1-CR3022, indicating that the primary sequence of those antibodies also plays some role. Instead, short-range van der Waals interactions with the inert gold surface cause a higher degree tilting and fraying of the antibodies with respect to amorphous silica. The interactions between the antibodies and the surface also affect the correlation among the different angles and dihedrals, which increases with their strength. Overall, results explain why amorphous silica substrates are frequently used to immobilize antibodies in immunosensors. © 2022 The Authors

JTD Keywords: amorphous silica, antibody immobilization, enzyme, gol d, gold, immobilization, immunosensor, molecu l a r dynamics, molecular dynamics, protein adsorption, sars-cov-2 immunosensor, simulations, spike protein, surface interactions, target, vaccine, Amorphous silica, Antibodies, Antibody engineering, Antibody immobilization, Antibody structure, Article, Chemical detection, Computer model, Controlled study, Dihedral angle, Gold, In-silico, Molecular dynamics, Molecular levels, Molecular-dynamics, Nonhuman, Property, Sars, Sars-cov-2 immunosensor, Severe acute respiratory syndrome coronavirus 2, Silica, Silico studies, Silicon dioxide, Solid substrates, Structure analysis, Substrate chemistry, Substrates, Van der waals forces, Virus detection

There has been a renewed interest in the potential use of psychedelics for the treatment of psychiatric conditions. Nevertheless, little is known about the mechanism of action and molecular pathways influenced by ayahuasca use in humans. Therefore, for the first time, our study aims to investigate the human metabolomics signature after consumption of a psychedelic, ayahuasca, and its connection with both the psychedelic-induced subjective effects and the plasma concentrations of ayahuasca alkaloids. Plasma samples of 23 individuals were collected both before and after ayahuasca consumption. Samples were analysed through targeted metabolomics and further integrated with subjective ratings of the ayahuasca experience (i.e., using the 5-Dimension Altered States of Consciousness Rating Scale [ASC]), and plasma ayahuasca-alkaloids using integrated network analysis. Metabolic pathways enrichment analysis using diffusion algorithms for specific KEGG modules was performed on the metabolic output. Compared to baseline, the consumption of ayahuasca increased N-acyl-ethanolamine endocannabinoids, decreased 2-acyl-glycerol endocannabinoids, and altered several large-neutral amino acids (LNAAs). Integrated network results indicated that most of the LNAAs were inversely associated with 9 out of the 11 subscales of the ASC, except for tryptophan which was positively associated. Several endocannabinoids and hexosylceramides were directly associated with the ayahuasca alkaloids. Enrichment analysis confirmed dysregulation in several pathways involved in neurotransmission such as serotonin and dopamine synthesis. In conclusion, a crosstalk between the circulating LNAAs and the subjective effects is suggested, which is independent of the alkaloid concentrations and provides insights into the specific metabolic fingerprint and mechanism of action underlying ayahuasca experiences. © 2022 The Authors

JTD Keywords: anxiety, ayahuasca, dimethyltryptamine, integrative network analysis, metabolism, metabolomics, psychedelics, rats, subjective effects, system, tryptophan, Ayahuasca, Dimethyltryptamine, Integrative network analysis, Metabolomics, Psychedelics, Serotonin 5-ht2a, Subjective effects

Biobased epoxy-derived raw materials will be essential for future coating and adhesive designs in industry. Here, a facile approach is reported towards the incorporation of limonene into an epoxy-functionalized polycarbonate and its crosslinking with a polyamine curing agent to obtain a thermoset material. For the first time, a solvent-borne adhesive with excellent film-forming, mechanical and adhesion strength properties is described.

JTD Keywords: adhesives, biobased epoxies, limonene, polycarbonate, Adhesives, Biobased epoxies, Biobased epoxy, Carbon-dioxide, Curing agents, Design in industries, Epoxides, Epoxy, Epoxy resins, Film adhesion, Film-forming, Functionalized, Limonene, Mechanical, Monomer, Monoterpenes, Oil, Oxide, Performance, Polyamines, Polycarbonate, Polycarbonates, Terpenes, Thermoset materials, Thermosets

Poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) enhance the delivery of therapeutic enzymes for replacement therapy of lysosomal storage disorders. Previous studies examined NPs encapsulating or coated with enzymes, but these formulations have never been compared. We examined this using hyaluronidase (HAse), deficient in mucopolysaccharidosis IX, and acid sphingomyelinase (ASM), deficient in types A–B Niemann–Pick disease. Initial screening of size, PDI, ζ potential, and loading resulted in the selection of the Lactel II co-polymer vs. Lactel I or Resomer, and Pluronic F68 surfactant vs. PVA or DMAB. Enzyme input and addition of carrier protein were evaluated, rendering NPs having, e.g., 181 nm diameter, 0.15 PDI, −36 mV ζ potential, and 538 HAse molecules encapsulated per NP. Similar NPs were coated with enzyme, which reduced loading (e.g., 292 HAse molecules/NP). NPs were coated with targeting antibodies (> 122 molecules/NP), lyophilized for storage without alterations, and acceptably stable at physiological conditions. NPs were internalized, trafficked to lysosomes, released active enzyme at lysosomal conditions, and targeted both peripheral organs and the brain after i.v. administration in mice. While both formulations enhanced enzyme delivery compared to free enzyme, encapsulating NPs surpassed coated counterparts (18.4- vs. 4.3-fold enhancement in cells and 6.2- vs. 3-fold enhancement in brains), providing guidance for future applications.

JTD Keywords: active enzymes, encapsulation, enhanced delivery, enzyme therapeutics, formulation parameters, icam-1 targeting, icam-1-targeted nanocarriers, in vivo biodistribution, in-vitro, lysosomal delivery, model, oral delivery, plga nanoparticles, poly(lactic-co-glycolic acid) nanoparticles, protein therapeutics, surface loading, Acid sphingomyelinase, Animals, Encapsulation, Enzyme replacement therapy, Enzyme therapeutics, Icam-1 targeting, In vivo biodistribution, Lysosomal delivery, Lysosomes, Mice, Nanoparticles, Poly(lactic-co-glycolic acid) nanoparticles, Polymers, Surface loading, Surface-active agents

Endothelial coverage of an exposed cardiovascular stent surface leads to the occurrence of restenosis and late-stent thrombosis several months after implantation. To overcome this difficulty, modification of stent surfaces with topographical or biochemical features may be performed to increase endothelial cells’ (ECs) adhesion and/or migration. This work combines both strategies on cobalt-chromium (CoCr) alloy and studies the potential synergistic effect of linear patterned surfaces that are obtained by direct laser interference patterning (DLIP), coupled with the use of Arg-Gly-Asp (RGD) and Tyr-Ile-Gly-Ser-Arg (YIGSR) peptides. An extensive characterization of the modified surfaces was performed by using AFM, XPS, surface charge, electrochemical analysis and fluorescent methods. The biological response was studied in terms of EC adhesion, migration and proliferation assays. CoCr surfaces were successfully patterned with a periodicity of 10 µm and two different depths, D (≈79 and 762 nm). RGD and YIGSR were immobilized on the surfaces by CPTES silanization. Early EC adhesion was increased on the peptide-functionalized surfaces, especially for YIGSR compared to RGD. High-depth patterns generated 80% of ECs’ alignment within the topographical lines and enhanced EC migration. It is noteworthy that the combined use of the two strategies synergistically accelerated the ECs’ migration and proliferation, proving the potential of this strategy to enhance stent endothelialization.

JTD Keywords: adhesion, bare-metal, biofunctionalization, biomaterials, cell adhesive peptides, cobalt-chromium alloy, direct laser interference patterning (dlip), endothelial cell migration, functionalization, matrix, proliferation, selectivity, shear-stress, surfaces, Biofunctionalization, Cell adhesive peptides, Cobalt-chromium alloy, Direct laser interference patterning (dlip), Drug-eluting stents, Endothelial cell migration

Many neurodegenerative disorders display protein aggregation as a hallmark, Huntingtin and TDP-43 aggregates being characteristic of Huntington disease and amyotrophic lateral sclerosis, respectively. However, whether these aggregates cause the diseases, are secondary by-products, or even have protective effects, is a matter of debate. Mutations in both human proteins can modulate the structure, number and type of aggregates, as well as their toxicity. To study the role of protein aggregates in cellular fitness, we have expressed in a highly tractable unicellular model different variants of Huntingtin and TDP-43. They each display specific patterns of aggregation and toxicity, even though in both cases proteins have to be very highly expressed to affect cell fitness. The aggregation properties of Huntingtin, but not of TDP-43, are affected by chaperones such as Hsp104 and the Hsp40 couple Mas5, suggesting that the TDP-43, but not Huntingtin, derivatives have intrinsic aggregation propensity. Importantly, expression of the aggregating form of Huntingtin causes a significant extension of fission yeast lifespan, probably as a consequence of kidnapping chaperones required for maintaining stress responses off. Our study demonstrates that in general these prion-like proteins do not cause toxicity under normal conditions, and in fact they can protect cells through indirect mechanisms which up-regulate cellular defense pathways. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.

JTD Keywords: aggregation, antioxidant, degradation, features, fission yeast, gene, huntingtin, neurodegenerative diseases, pap1, polyglutamine toxicity, protein aggregation, proteins, stress, tdp-43, Amyotrophic-lateral-sclerosis, Chaperone, Chemistry, Dna binding protein, Dna-binding proteins, Fission yeast, Genetics, Human, Humans, Huntingtin, Metabolism, Molecular chaperones, Neurodegenerative diseases, Prion, Prions, Protein aggregate, Protein aggregates, Protein aggregation, Schizosaccharomyces, Tdp-43

Decellularization procedures have been developed and optimized for the entire organ or tissue blocks, by either perfusion of decellularizing agents through the tissue’s vasculature or submerging large sections in decellularizing solutions. However, some research aims require the analysis of native as well as decellularized tissue slices side by side, but an optimal protocol has not yet been established to address this need. Thus, the main goal of this work was to develop a fast and efficient decellularization method for tissue slices—with an emphasis on lung—while attached to a glass slide. To this end, different decellularizing agents were compared for their effectiveness in cellular removal while preserving the extracellular matrix. The intensity of DNA staining was taken as an indicator of remaining cells and compared to untreated sections. The presence of collagen, elastin and laminin were quantified using immunostaining and signal quantification. Scaffolds resulting from the optimized protocol were mechanically characterized using atomic force microscopy. Lung scaffolds were recellularized with mesenchymal stromal cells to assess their biocompatibility. Some decellularization agents (CHAPS, triton, and ammonia hydroxide) did not achieve sufficient cell removal. Sodium dodecyl sulfate (SDS) was effective in cell removal (1% remaining DNA signal), but its sharp reduction of elastin signal (only 6% remained) plus lower attachment ratio (32%) singled out sodium deoxycholate (SD) as the optimal treatment for this application (6.5% remaining DNA signal), due to its higher elastin retention (34%) and higher attachment ratio (60%). Laminin and collagen were fully preserved in all treatments. The SD decellularization protocol was also successful for porcine and murine (mice and rat) lungs as well as for other tissues such as the heart, kidney, and bladder. No significant mechanical differences were found before and after sample decellularization. The resulting acellular lung scaffolds were shown to be biocompatible (98% cell survival after 72 h of culture). This novel method to decellularize tissue slices opens up new methodological possibilities to better understand the role of the extracellular matrix in the context of several diseases as well as tissue engineering research and can be easily adapted for scarce samples like clinical biopsies. Copyright © 2022 Narciso, Ulldemolins, Júnior, Otero, Navajas, Farré, Gavara and Almendros.

JTD Keywords: biocompatibility, bioscaffold recellularization, decellularization, extracellular matrix, flow, impact, lung, scaffolds, tissue slices, Ammonia, Bio-scaffolds, Biocompatibility, Biological organs, Bioscaffold recellularization, Cell removal, Cells, Collagen, Cytology, Decellularization, Dna, Dna signals, Elastin, Extracellular matrices, Extracellular matrix, Extracellular-matrix, Glycoproteins, Laminin, Lung, Mammals, Recellularization, Scaffolds (biology), Sodium deoxycholate, Sulfur compounds, Tissue, Tissue slice, Tissue slices

In the area of coating development, it is extremely difficult to find a substitute for bisphenol A diglycidyl ether (DGEBA), the classical petroleum-based raw material used for the formulation of epoxy thermosets. This epoxy resin offers fast curing reaction with several hardeners and the best thermal and chemical resistance properties for applications in coatings and adhesive technologies. In this work, a new biobased epoxy, derived from poly(limonene carbonate) oxide (PLCO), was combined with polyetheramine and polyamineamide curing agents, offering a spectrum of thermal and mechanical properties, superior to DGEBA-based thermosets. The best formulation was found to be a combination of PLCO and a commercial curing agent (Jeffamine) in a stoichiometric 1:1 ratio. Although PLCO is a solid due to its high molecular weight, it was possible to create a two-component partially biobased epoxy paint without the need of volatile organic compounds (i.e., solvent-free formulation), intended for use in coating technology to partially replace DGEBA-based thermosets.

JTD Keywords: acid, adhesion, epoxy thermoset, mechanical properties, monomer, polycarbonates, polymers, protection, resins, solvent-free paint, thermal properties, Adhesives, Biobased epoxy, Bisphenol-a-diglycidyl ethers, Carbonation, Coating development, Coating technologies, Curing, Curing agents, Epoxy coatings, Epoxy resins, Epoxy thermoset, Epoxy thermosets, Limonene oxide, Mechanical properties, Monoterpenes, Paint, Poly(limonene carbonate) oxide, Solvent free, Solvent-free paint, Thermal properties, Thermosets, Volatile organic compounds

The endosomal sorting complex required for transport (ESCRT) machinery mediates membrane fission reactions that exhibit a different topology from that observed in clathrin-coated vesicles. In all of the ESCRT-mediated events, the nascent vesicle buds away from the cytosol. However, ESCRT proteins are able to act upon membranes with different geometries. For instance, the formation of multivesicular bodies (MVBs) and the biogenesis of extracellular vesicles both require the participation of the ESCRT-III sub-complex, and they differ in their initial membrane geometry before budding starts: the protein complex acts either from outside the membrane organelle (causing inward budding) or from within (causing outward budding). Several studies have reconstituted the action of the ESCRT-III subunits in supported bilayers and cell-sized vesicles mimicking the geometry occurring during MVBs formation (in-bud), but extracellular vesicle budding (out-bud) mechanisms remain less explored, because of the outstanding difficulties encountered in encapsulation of functional ESCRT-III in vesicles. Here, we provide a different approach that allows the recreation of the out-bud formation, by combining giant unilamellar vesicles as a membrane model and a microinjection system. The vesicles are immobilized prior to injection via weak adhesion to the chamber coverslip, which also ensures preserving the membrane excess area required for budding. After protein injection, vesicles exhibit outward budding. The approach presented in this work can be used in the future to disentangle the mechanisms underlying ESCRT-III-mediated fission, recreating the geometry of extracellular bud production, which remains a challenge. Moreover, the microinjection methodology can be also adapted to interrogate the action of other cytosolic components on the encapsulating membranous organelle. Copyright: © 2022 The Authors.

JTD Keywords: adhesion, budding, electroformation, escrt-iii, exosomes, extracellular vesicles, giant unilamellar vesicle (guv), light, microinjection, microparticles, plasma, Adhesion, Budding, Escrt-iii, Extracellular vesicles, Giant unilamellar vesicle (guv), Membrane, Microinjection

JTD Keywords: augmentation, calcium-phosphate, expansion, fabrication, hydroxyapatite, made artificial bones, osteogenesis, reconstruction, regeneration, 3-d printing, 3d printers, 3d printing, 3d-printing, Abstracting, Additives, Biomaterial, Bone, Bone regeneration, Bone substitution, Bone tissue engineering, Ceramic, Ceramics, Clinic, Controlled drug delivery, Personalized medicines, Surgical planning, Targeted drug delivery, Three-dimensional-printing, Tricalcium phosphate scaffolds

The impact of diet and digestive disorders in flatus composition remains largely unexplored. This is partially due to the lack of standardized sampling collection methods, and the easy atmospheric contamination. This paper describes a method to quantitatively determine the major gases in flatus and their application in a nutritional intervention. We describe how to direct sample flatus into Tedlar bags, and simultaneous analysis by gas chromatography–thermal conductivity detection (GC–TCD). Results are analyzed by univariate hypothesis testing and by multilevel principal component analysis. The reported methodology allows simultaneous determination of the five major gases with root mean measurement errors of 0.8% for oxygen (O2), 0.9% for nitrogen (N2), 0.14% for carbon dioxide (CO2), 0.11% for methane (CH4), and 0.26% for hydrogen (H2). The atmospheric contamination was limited to 0.86 (95% CI: [0.7–1.0])% for oxygen and 3.4 (95% CI: [1.4–5.3])% for nitrogen. As an illustration, the method has been successfully applied to measure the response to a nutritional intervention in a reduced crossover study in healthy subjects. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.

JTD Keywords: breath, colonic microbiota, diet effect on flatus, disorders, evacuation, excretion, flatulence, hydrogen gas, major flatus gas components, multilevel principal component analysis, rectal gas collection, systems, volume, Atmospheric contamination, Carbon dioxide, Conductivity detection, Diet effect on flatus, Gas chromatography, Gas collections, Gas component, Gases, Major flatus gas component, Major flatus gas components, Multilevel principal component analyse, Multilevel principal component analysis, Multilevels, Nitrogen, Nutrition, Oxygen, Principal component analysis, Principal-component analysis, Rectal gas collection, Volatile organic-compounds

JTD Keywords: amino-acids, catalysis, dopant-free hydroxyapatite, electrical properties, electrophotosynthesis, nitrogen, thermally-stimulated polarization, Advanced materials, Biocompatibility, Biomedical applications, Brushite, Doped hydroxyapatites, Electric voltage, Electrical characterization, Electrochemical impedance spectroscopy, Equivalent circuits, Future perspectives, Highest temperature, Hydroxyapatite, Interfaces (materials), Material interfaces, Medical applications, Polarization, Polarization conditions, Surface-charges, Technological applications

Underpowered trials risk inaccurate results. Recruitment to stroke rehabilitation randomised controlled trials (RCTs) is often a challenge. Statistical simulations offer an important opportunity to explore the adequacy of sample sizes in the context of specific outcome measures. We aimed to examine and compare the adequacy of stroke rehabilitation RCT sample sizes using the Barthel Index (BI) or modified Rankin Scale (mRS) as primary outcomes.We conducted computer simulations using typical experimental event rates (EER) and control event rates (CER) based on individual participant data (IPD) from stroke rehabilitation RCTs. Event rates are the proportion of participants who experienced clinically relevant improvements in the RCT experimental and control groups. We examined minimum sample size requirements and estimated the number of participants required to achieve a number needed to treat within clinically acceptable boundaries for the BI and mRS.We secured 2350 IPD (18 RCTs). For a 90% chance of statistical accuracy on the BI a rehabilitation RCT would require 273 participants per randomised group. Accurate interpretation of effect sizes would require 1000s of participants per group. Simulations for the mRS were not possible as a clinically relevant improvement was not detected when using this outcome measure.Stroke rehabilitation RCTs with large sample sizes are required for accurate interpretation of effect sizes based on the BI. The mRS lacked sensitivity to detect change and thus may be unsuitable as a primary outcome in stroke rehabilitation trials.Copyright © 2021 Elsevier Inc. All rights reserved.

JTD Keywords:  , barthel index, design, increasing value, modified rankin scale, randomised controlled trials, recruitment, reducing waste, reliability, sample size calculations, simulations, stroke rehabilitation, Adult, Article, Barthel index, Calculation, Computer simulation, Controlled study, Effect size, Female, Human, Human experiment, Major clinical study, Male, Modified rankin scale, Numbers needed to treat, Outcome assessment, Randomised controlled trials, Randomized controlled trial, Randomized controlled-trials, Rankin scale, Recruitment, Rehabilitation, Sample size, Sample size calculations, Simulations, Stroke rehabilitation

[No abstract available]

JTD Keywords: Adult, Aged, Female, Hemiplegia, Humans, Male, Middle aged, Recovery of function, Rehabilitation, Retrospective studies, Stroke, Stroke recovery, Stroke rehabilitation

Unlike the attachment of soft epithelial skin tissue to penetrating solid natural structures like fingernails and teeth, sealing around percutaneous/permucosal devices such as dental implants is hindered by inflammation and epidermal down growth. Here, we employed a dual keratinocyte-adhesive peptide and anti-inflammatory biomolecule coating on titanium to promote oral epithelial tissue attachment. For minimizing inflammation-triggered epidermal down growth, we coated pristine and oxygen plasma pre-treated polished titanium (pTi) with conjugated linoleic acid (CLA). Further, in order to aid in soft tissue attachment via the formation of hemidesmosomes, adhesive structures by oral keratinocytes, we coated the anionic linoleic acid (LA) adsorbed titanium with cationic cell adhesive peptides (CAP), LamLG3, a peptide derived from Laminin 332, the major extracellular matrix component of the basement membrane in skin tissue and Net1, derived from Netrin-1, a neural chemoattractant capable of epithelial cell attachment via alpha 6 beta 4 integrins. The dual CLA-CAP coatings on pTi were characterized by X-ray photoelectron spectroscopy and dynamic water contact angle measurements. The proliferation of human oral keratinocytes (TERT-2/OKF6) was accelerated on the peptide coated titanium while also promoting the expression of Col XVII and beta-4 integrin, two markers for hemidesmosomes. Simultaneously, CLA coating suppressed the production of inducible nitric oxide synthase (anti-iNOS); a pro-inflammatory M1 marker expressed in lipopolysaccharide (LPS) stimulated murine macrophages (RAW 264.7) and elevated expression of anti-CD206, associated to an anti-inflammatory M2 macrophage phenotype. Taken together, the dual keratinocyte-adhesive peptide and anti-inflammatory biomolecule coating on titanium can help reduce inflammation and promote permucosal/peri-implant soft tissue sealing.

JTD Keywords: Adhesives, Animal, Animals, Anti-inflammatories, Anti-inflammatory agents, Antiinflammatory agent, Biomolecules, Bone, Cell adhesion, Cell-adhesives, Coatings, Conjugated linoleic acid, Conjugated linoleic-acid, Contact angle, Hemidesmosome, Hemidesmosomes, Human, Humans, Hydroxyapatite, Inflammation, Integrins, Keratinocyte, Keratinocytes, Linoleic acid, Macrophages, Mice, Mouse, Nitric oxide, Oral implants, Pathology, Peptides, Skin tissue, Soft tissue, Supplementation, Surface properties, Surface property, Tissue, Titania, Titanium, X ray photoelectron spectroscopy

Quantum dots (QDs) are semiconductor nanoparticles with unique optical and electronic properties, whose interest as potential nano-theranostic platforms for imaging and sensing is increasing. The design and use of QDs requires the understanding of cell-nanoparticle interactions at a microscopic and nanoscale level. Model systems such as supported lipid bilayers (SLBs) are useful, less complex platforms mimicking physico-chemical properties of cell membranes. In this work, we investigated the effect of topographical homogeneity of SLBs bearing different surface charge in the adsorption of hydrophilic QDs. Using quartz-crystal microbalance, a label-free surface sensitive technique, we show significant differences in the interactions of QDs onto homogeneous and inhomogeneous SLBs formed following different strategies. Within short time scales, QDs adsorb onto topographically homogeneous, defect-free SLBs is driven by electrostatic interactions, leading to no layer disruption. After prolonged QD exposure, the nanomechanical stability of the SLB decreases suggesting nanoparticle insertion. In the case of inhomogeneous, defect containing layers, QDs target preferentially membrane defects, driven by a subtle interplay of electrostatic and entropic effects, inducing local vesicle rupture and QD insertion at membrane edges. © 2021

JTD Keywords: adsorption, atomic force microscopy, bilayer formation, gold nanoparticles, hydrophilic quantum dots, lipid membrane defects, model, nanomechanics, quartz crystal microbalance with dissipation, size, supported lipid bilayers, surfaces, Atomic force microscopy, Atomic-force-microscopy, Cell membrane, Cytology, Defect-free, Electronic properties, Electrostatics, Hydrophilic quantum dot, Hydrophilic quantum dots, Hydrophilicity, Hydrophilics, Hydrophobic and hydrophilic interactions, Lipid bilayers, Lipid membrane defect, Lipid membrane defects, Lipid membranes, Lipids, Nanocrystals, Nanomechanics, Optical and electronic properties, Quantum dots, Quartz, Quartz crystal microbalance techniques, Quartz crystal microbalance with dissipation, Quartz crystal microbalances, Quartz-crystal microbalance, Semiconductor nanoparticles, Semiconductor quantum dots, Supported lipid bilayers

Despite tremendous progress in the understanding of COVID-19, mechanistic insight into immunological, disease-driving factors remains limited. We generated maVie16, a mouse-adapted SARS-CoV-2, by serial passaging of a human isolate. In silico modeling revealed how only three Spike mutations of maVie16 enhanced interaction with murine ACE2. maVie16 induced profound pathology in BALB/c and C57BL/6 mice, and the resulting mouse COVID-19 (mCOVID-19) replicated critical aspects of human disease, including early lymphopenia, pulmonary immune cell infiltration, pneumonia, and specific adaptive immunity. Inhibition of the proinflammatory cyto-kines IFN? and TNF substantially reduced immunopathology. Importantly, genetic ACE2-deficiency completely prevented mCOVID-19 development. Finally, inhalation therapy with recombinant ACE2 fully protected mice from mCOVID-19, revealing a novel and efficient treatment. Thus, we here present maVie16 as a new tool to model COVID-19 for the discovery of new therapies and show that disease severity is determined by cytokine-driven immunopathology and critically dependent on ACE2 in vivo. © Gawish et al.

JTD Keywords: covid-19 mouse model, covid-19 therapy, cytokine storm, immunology, inflammation, mavie16, mouse, mouse-adapted sars-cov-2, program, recombinant soluble ace2, tmprss2, Adaptive immunity, Angiotensin converting enzyme 2, Angiotensin-converting enzyme 2, Animal, Animal cell, Animal experiment, Animal model, Animal tissue, Animals, Apoptosis, Article, Bagg albino mouse, Breathing rate, Bronchoalveolar lavage fluid, C57bl mouse, Cell composition, Cell infiltration, Controlled study, Coronavirus disease 2019, Coronavirus spike glycoprotein, Covid-19, Cytokeratin 18, Cytokine production, Dipeptidyl carboxypeptidase, Disease model, Disease models, animal, Disease severity, Drosophila-melanogaster, Enzyme linked immunosorbent assay, Expression vector, Flow cytometry, Gamma interferon, Gene editing, Gene expression, Gene mutation, Genetic engineering, Genetics, Glycosylation, High mobility group b1 protein, Histology, Histopathology, Immune response, Immunocompetent cell, Immunology, Immunopathology, Interferon-gamma, Interleukin 2, Metabolism, Mice, inbred balb c, Mice, inbred c57bl, Mouse-adapted sars-cov-2, Myeloperoxidase, Neuropilin 1, Nonhuman, Nucleocapsid protein, Pathogenicity, Peptidyl-dipeptidase a, Pyroptosis, Recombinant soluble ace2, Renin angiotensin aldosterone system, Rna extraction, Rna isolation, Sars-cov-2, Severe acute respiratory syndrome coronavirus 2, Spike glycoprotein, coronavirus, T lymphocyte activation, Trabecular meshwork, Tumor necrosis factor, Virology, Virus load, Virus replication, Virus transmission, Virus virulence

Several studies have demonstrated the different characteristics of tau seeding and spreading following intracerebral inoculation in murine models of tau-enriched fractions of brain homogenates from AD and other tauopathies. The present study is centered on the importance of host tau in tau seeding and the molecular changes associated with the transformation of host tau into abnormal tau. The brains of three adult murine genotypes expressing different forms of tau—WT (murine 4Rtau), hTau (homozygous transgenic mice knock-out for murine tau protein and heterozygous expressing human forms of 3Rtau and 4Rtau proteins), and mtWT (homozygous transgenic mice knock-out for murine tau protein)—were analyzed following unilateral hippocampal inoculation of sarkosyl-insoluble tau fractions from the same AD and control cases. The present study reveals that (a) host tau is mandatory for tau seeding and spreading following tau inoculation from sarkosyl-insoluble fractions obtained from AD brains; (b) tau seeding does not occur following intracerebral inoculation of sarkosyl-insoluble fractions from controls; (c) tau seeding and spreading are characterized by variable genotype-dependent tau phosphorylation and tau nitration, MAP2 phosphorylation, and variable activation of kinases that co-localize with abnormal tau deposits; (d) transformation of host tau into abnormal tau is an active process associated with the activation of specific kinases; (e) tau seeding is accompanied by modifications in tau splicing, resulting in the expression of new 3Rtau and 4Rtau isoforms, thus indicating that inoculated tau seeds have the capacity to model exon 10 splicing of the host mapt or MAPT with a genotype-dependent pattern; (e) selective regional and cellular vulnerabilities, and different molecular compositions of the deposits, are dependent on the host tau of mice injected with identical AD tau inocula.

JTD Keywords: 3rtau and 4rtau, alzheimer's disease, alzheimer’s disease, brains, granulovacuolar degeneration, host tau, htau, intranuclear distribution, messenger-rna, pathological tau, propagation, protein-kinases, seeding and spreading, tauopathies, transmission, 3rtau and 4rtau, Alzheimer disease, Alzheimers-disease, Alzheimer’s disease, Animals, Biomarkers, Brain, Disease models, animal, Disease susceptibility, Fluorescent antibody technique, Genotype, Hippocampus, Host tau, Htau, Humans, Immunohistochemistry, Mice, Mice, knockout, Mice, transgenic, Mutation, Neurons, Seeding and spreading, Tau proteins, Tauopathies

Maintaining a balance between excitatory and inhibitory activity is an essential feature of neural networks of the neocortex. In the face of perturbations in the levels of excitation to cortical neurons, synapses adjust to maintain excitatory-inhibitory (EI) balance. In this review, we summarize research on this EI homeostasis in the neocortex, using stroke as our case study, and in particular the loss of excitation to distant cortical regions after focal lesions. Widespread changes following a localized lesion, a phenomenon known as diaschisis, are not only related to excitability, but also observed with respect to functional connectivity. Here, we highlight the main findings regarding the evolution of excitability and functional cortical networks during the process of post-stroke recovery, and how both are related to functional recovery. We show that cortical reorganization at a global scale can be explained from the perspective of EI homeostasis. Indeed, recovery of functional networks is paralleled by increases in excitability across the cortex. These adaptive changes likely result from plasticity mechanisms such as synaptic scaling and are linked to EI homeostasis, providing a possible target for future therapeutic strategies in the process of rehabilitation. In addition, we address the difficulty of simultaneously studying these multiscale processes by presenting recent advances in large-scale modeling of the human cortex in the contexts of stroke and EI homeostasis, suggesting computational modeling as a powerful tool to tie the meso- and macro-scale processes of recovery in stroke patients. Copyright © 2022 Páscoa dos Santos and Verschure.

JTD Keywords: balanced excitation, canonical microcircuit, cerebral-cortex, cortical excitability, cortical reorganization, diaschisis, excitability, excitatory-inhibitory balance, functional networks, homeostatic plasticity, ischemic-stroke, neuronal avalanches, photothrombotic lesions, state functional connectivity, whole-brain models, Algorithm, Biological marker, Brain, Brain cell, Brain cortex, Brain function, Brain radiography, Cerebrovascular accident, Cortical reorganization, Diaschisis, Down regulation, Excitability, Excitatory-inhibitory balance, Fluorine magnetic resonance imaging, Functional networks, Homeostasis, Homeostatic plasticity, Human, Motor dysfunction, Neuromodulation, Plasticity, Pyramidal nerve cell, Review, Simulation, Stroke, Stroke patient, Theta-burst stimulation, Visual cortex

Different copolymers incorporating terpene oxide units (e.g., limonene oxide) have been evaluated considering thermal properties, degradability, and biocompatibility. Thus, polycarbonates and polyesters derived from aromatic, monocyclic and bicyclic anhydrides have been considered. Furthermore, ring substitution with myrcene terpene has been evaluated. All polymers were amorphous when evaluated directly from synthesis. However, spherulites could be observed after the slow evaporation of diluted chloroform solutions of polylimonene carbonate, with all isopropene units possessing an R configuration. This feature was surprising considering the reported information that suggested only the racemic polymer was able to crystallize. All polymers were thermally stable and showed a dependence of the maximum degradation rate temperature (from 242 °C to 342 °C) with the type of terpene oxide. The graduation of glass transition temperatures (from 44 °C to 172 °C) was also observed, being higher than those corresponding to the unsubstituted polymers. The chain stiffness of the studied polymers hindered both hydrolytic and enzymatic degradation while a higher rate was detected when an oxidative medium was assayed (e.g., weight losses around 12% after 21 days of exposure). All samples were biocompatible according to the adhesion and proliferation tests performed with fibroblast cells. Hydrophobic and mechanically consistent films (i.e., contact angles between 90° and 110°) were obtained after the evaporation of chloroform from the solutions, having different ratios of the studied biobased polyterpenes and poly(butylene succinate) (PBS). The blend films were comparable in tensile modulus and tensile strength with the pure PBS (e.g., values of 330 MPa and 7 MPa were determined for samples incorporating 30 wt.% of poly(PA-LO), the copolyester derived from limonene oxide and phthalic anhydride. Blends were degradable, biocompatible and appropriate to produce oriented-pore and random-pore scaffolds via a thermally-induced phase separation (TIPS) method and using 1,4-dioxane as solvent. The best results were attained with the blend composed of 70 wt.% PBS and 30 wt.% poly(PA-LO). In summary, the studied biobased terpene derivatives showed promising properties to be used in a blended form for biomedical applications such as scaffolds for tissue engineering.

JTD Keywords: alternating copolymerization, biobased materials, biodegradability, composites, crystallization, cyclohexene oxide, induced phase-separation, limonene oxide, mechanical-properties, polyesters, scaffolds, spherulites, terpene derivatives, thermal properties, thermally-induced phase separation, Acetone, Bio-based, Bio-based materials, Biobased materials, Biocompatibility, Biodegradability, Butenes, Cell culture, Chlorine compounds, Degradation, Evaporation, Glass transition, Limonene oxide, Monoterpenes, Phase separation, Poly (butylenes succinate), Polybutylene succinate, Property, Ring-opening copolymerization, Scaffolds, Spheru-lites, Tensile strength, Terpene derivatives, Thermal properties, Thermally induced phase separation, Thermally-induced phase separation, Thermally?induced phase separation, Thermodynamic properties, Thermogravimetric analysis

MicroRNAs (miRNAs) are small non-coding endogenous RNAs, which are attracting a growing interest as therapeutic molecules due to their central role in major diseases. However, the transformation of these biomolecules into drugs is limited due to their unstability in the bloodstream, caused by nucleases abundantly present in the blood, and poor capacity to enter cells. The conjugation of miRNAs to nanoparticles (NPs) could be an effective strategy for their clinical delivery. Herein, the engineering of non-liposomal lipid nanovesicles, named quatsomes (QS), for the delivery of miRNAs and other small RNAs into the cytosol of tumor cells, triggering a tumor-suppressive response is reported. The engineered pH-sensitive nanovesicles have controlled structure (unilamellar), size (<150 nm) and composition. These nanovesicles are colloidal stable (>24 weeks), and are prepared by a green, GMP compliant, and scalable one-step procedure, which are all unavoidable requirements for the arrival to the clinical practice of NP based miRNA therapeutics. Furthermore, QS protect miRNAs from RNAses and when injected intravenously, deliver them into liver, lung, and neuroblastoma xenografts tumors. These stable nanovesicles with tunable pH sensitiveness constitute an attractive platform for the efficient delivery of miRNAs and other small RNAs with therapeutic activity and their exploitation in the clinics.

JTD Keywords: cancer therapy, mirnas delivery, nanocarriers, nanovesicles, neuroblastoma, pediatric cancer, quatsomes, Biodistribution, Cancer therapy, Cell engineering, Cells, Cholesterol, Controlled drug delivery, Diseases, Dna, Dysregulated ph, Lipoplex, Microrna delivery, Mirnas delivery, Nanocarriers, Nanoparticles, Nanovesicle, Nanovesicles, Neuroblastoma, Neuroblastomas, Pediatric cancer, Ph sensitive, Ph sensors, Quatsome, Quatsomes, Rna, Sirna, Sirna delivery, Sirnas delivery, Small interfering rna, Small rna, Targeted drug delivery, Tumors, Vesicles

The recovery of motor function after suffering a brain injury is still poorly understood. There are complex interactions between motor learning and motor recovery principles, which guide neural reorganization and allow the reacquisition and restitution of goal-oriented motor patterns (Maier et al. in Front. Syst. Neurosci. 13:74 (2019) [1]). This paper, reviews several studies and applications for rehabilitation and impairment assessment. Were we capitalize on the interaction between motor learning and motor recovery principles and investigate the manipulation of visuomotor feedback for the delivery of optimal neurorehabilitation and diagnostics for motor deficits after stroke and cannabis addiction.

JTD Keywords: Adaptation, Good health and well-being

Mycobacterium bovis bacillus Calmette-Guérin (BCG) efficacy as an immunotherapy tool can be influenced by the genetic background or immune status of the treated population and by the BCG substrain used. BCG comprises several substrains with genetic differences that elicit diverse phenotypic characteristics. Moreover, modifications of phenotypic characteristics can be influenced by culture conditions. However, several culture media formulations are used worldwide to produce BCG. To elucidate the influence of growth conditions on BCG characteristics, five different substrains were grown on two culture media, and the lipidic profile and physico-chemical properties were evaluated. Our results show that each BCG substrain displays a variety of lipidic profiles on the outermost surface depending on the growth conditions. These modifications lead to a breadth of hydrophobicity patterns and a different ability to reduce neutral red dye within the same BCG substrain, suggesting the influence of BCG growth conditions on the interaction between BCG cells and host cells.

JTD Keywords: cell wall, efficacy, glycerol, hydrophobicity, lipid, neutral red, pdim, pgl, protein, strains, viability, virulence, Acylglycerol, Albumin, Article, Asparagine, Bacterial cell wall, Bacterial gene, Bacterium culture, Bcg vaccine, Catalase, Cell wall, Chloroform, Controlled study, Escherichia coli, Gene expression, Genomic dna, Glycerol, Glycerol monomycolate, Hexadecane, Housekeeping gene, Hydrophobicity, Immune response, Immunogenicity, Immunotherapy, Lipid, Lipid fingerprinting, Magnesium sulfate, Mercaptoethanol, Methanol, Methylglyoxal, Molybdatophosphoric acid, Mycobacterium bovis bcg, Neutral red, Nonhuman, Pdim, Petroleum ether, Pgl, Phenotype, Physical chemistry, Real time reverse transcription polymerase chain reaction, Rna 16s, Rna extraction, Rv0577, Staining, Thin layer chromatography, Unclassified drug

Introduction: After a stroke, a wide range of deficits can occur with varying onset latencies. As a result, assessing impairment and recovery are enormous challenges in neurorehabilitation. Although several clinical scales are generally accepted, they are time-consuming, show high inter-rater variability, have low ecological validity, and are vulnerable to biases introduced by compensatory movements and action modifications. Alternative methods need to be developed for efficient and objective assessment. In this study, we explore the potential of computer-based body tracking systems and classification tools to estimate the motor impairment of the more affected arm in stroke patients. Methods: We present a method for estimating clinical scores from movement parameters that are extracted from kinematic data recorded during unsupervised computer-based rehabilitation sessions. We identify a number of kinematic descriptors that characterise the patients' hemiparesis (e.g., movement smoothness, work area), we implement a double-noise model and perform a multivariate regression using clinical data from 98 stroke patients who completed a total of 191 sessions with RGS. Results: Our results reveal a new digital biomarker of arm function, the Total Goal-Directed Movement (TGDM), which relates to the patients work area during the execution of goal-oriented reaching movements. The model's performance to estimate FM-UE scores reaches an accuracy of R-2: 0.38 with an error (sigma: 12.8). Next, we evaluate its reliability (r = 0.89 for test-retest), longitudinal external validity (95% true positive rate), sensitivity, and generalisation to other tasks that involve planar reaching movements (R-2: 0.39). The model achieves comparable accuracy also for the Chedoke Arm and Hand Activity Inventory (R-2: 0.40) and Barthel Index (R-2: 0.35). Conclusions: Our results highlight the clinical value of kinematic data collected during unsupervised goal-oriented motor training with the RGS combined with data science techniques, and provide new insight into factors underlying recovery and its biomarkers.

JTD Keywords: interactive feedback, motion classification, motion sensing, multivariate regression, posture monitoring, rehabilitation, stroke, Adult, Aged, Analytic method, Arm movement, Article, Barthel index, Brain hemorrhage, Cerebrovascular accident, Chedoke arm and hand activity inventory, Clinical protocol, Cognitive defect, Computer analysis, Controlled study, Convergent validity, Correlation coefficient, Disease severity, External validity, Female, Fugl meyer assessment for the upper extremity, Functional assessment, Functional status assessment, General health status assessment, Hemiparesis, Human, Interactive feedback, Ischemic stroke, Kinematics, Major clinical study, Male, Mini mental state examination, Motion classification, Motion sensing, Motor analog scale, Movement, Multivariate regression, Muscle function, Posture monitoring, Probability, Recovery, Rehabilitation, Reliability, Retrospective study, Stroke, Stroke patient, Test retest reliability, Therapy, Total goal directed movement, Upper extremities, Upper limb, Upper-limb, Wolf motor function test

Polypeptide-based nanoparticles offer unique advantages from a nanomedicine perspective such as biocompatibility, biodegradability, and stimuli-responsive properties to (patho)physiological conditions. Conventionally, self-assembled polypeptide nanostructures are prepared by first synthesizing their constituent amphiphilic polypeptides followed by postpolymerization self-assembly. Herein, we describe the one-pot synthesis of oxidation-sensitive supramolecular micelles and vesicles. This was achieved by polymerization-induced self-assembly (PISA) of the N-carboxyanhydride (NCA) precursor of methionine using poly(ethylene oxide) as a stabilizing and hydrophilic block in dimethyl sulfoxide (DMSO). By adjusting the hydrophobic block length and concentration, we obtained a range of morphologies from spherical to wormlike micelles, to vesicles. Remarkably, the secondary structure of polypeptides greatly influenced the final morphology of the assemblies. Surprisingly, wormlike micellar morphologies were obtained for a wide range of methionine block lengths and solid contents, with spherical micelles restricted to very short hydrophobic lengths. Wormlike micelles further assembled into oxidation-sensitive, self-standing gels in the reaction pot. Both vesicles and wormlike micelles obtained using this method demonstrated to degrade under controlled oxidant conditions, which would expand their biomedical applications such as in sustained drug release or as cellular scaffolds in tissue engineering.

JTD Keywords: alpha-amino-acid, hydrogels, leuchs anhydrides, platform, polypeptides, transformation, triggered cargo release, Amino acids, Amphiphilics, Biocompatibility, Biodegradability, Block lengths, Controlled drug delivery, Dimethyl sulfoxide, Ethylene, Gels, Hydrophobicity, Medical nanotechnology, Methionine, Micelles, Morphology, Nanoparticles, One-pot synthesis, Organic solvents, Oxidation, Physiological condition, Polyethylene glycols, Polyethylene oxides, Polymerization, Post-polymerization, Ring-opening polymerization, Scaffolds (biology), Self assembly, Stimuli-responsive properties, Supramolecular chemistry, Supramolecular gels, Supramolecular micelles, Wormlike micelle

JTD Keywords: added value chemicals, amino-acids, catalytic-hydrogenation, climate, design, electrochemical reduction, electroreduction, green co2 conversion to ethanol, nitrogen, photocatalytic reduction, polarized hydroxyapatite, recycling co2, sea-level, Acetone, Added value chemicals, Added-value chemicals, C-c coupling, Calcium apatites, Carbon dioxide, Carbon-dioxide, Co 2 reduction, Co2 reduction, Ethanol, Green co2 conversion to ethanol, Hard tissues, Hydroxyapatite, Mixtures, Morphology, Morphology and composition, Naturally occurring, Organic carbon, Phosphate minerals, Polarized hydroxyapatite, Recycling co2

JTD Keywords: biocompatibility, bioresorbable stents, degradation, mechanical-properties, poly(epsilon-caprolactone), poly-l-lactic acid, polylactide, radiopacity, thermogel, x-ray imaging, Barium sulfate, Biocompatibility, Bioresorbable, Bioresorbable scaffolds, Bioresorbable stent, Bioresorbable stents, Blood vessels, Computerized tomography, Controlled drug delivery, Coronary heart disease, Direct-writing, Endothelial cells, Fabrication strategies, Injection molding, Lactic acid, Poly-l-lactic acid, Poly-l-lactic acids, Radiopacity, Scaffolds (biology), Solvent cast, Solvent-cast direct-writing, Solvents, Stents, Struts, Sulfur compounds, Targeted drug delivery, X-ray imaging

In the recent decades, zinc (Zn) and its alloys have been drawing attention as promising candidates for bioresorbable cardiovascular stents due to its degradation rate more suitable than magnesium (Mg) and iron (Fe) alloys. However, its mechanical properties need to be improved in order to meet the criteria for vascular stents. This work investigates the mechanical properties, biodegradability and biocompatibility of Zn-Mg and Zn-Cu alloys in order to determine a proper alloy composition for optimal stent performance. Nanoindentation measurements are performed to characterize the mechanical properties at the nanoscale as a function of the Zn microstructure variations induced by alloying. The biodegradation mechanisms are discussed and correlated to microstructure, mechanical performance and bacterial/cell response. Addition of Mg or Cu alloying elements refined the microstructure of Zn and enhanced yield strength (YS) and ultimate tensile strength (UTS) proportional to the volume fraction of secondary phases. Zn-1Mg showed the higher YS and UTS and better performance in terms of degradation stability in Hanks’ solution. Zn-Cu alloys presented an antibacterial effect for S. aureus controlled by diffusion mechanisms and by contact. Biocompatibility was dependent on the degradation rate and the nature of the corrosion products.

JTD Keywords: behavior, biocompatibility, biodegradable metals, bioresorbable metals, bioresorbable scaffold, copper, corrosion properties, elastic-modulus, galvanic corrosion, microstructure, nanoindentation, redox homeostasis, zinc, Biocompatibility, Bioresorbable metals, Galvanic corrosion, Nanoindentation, Room-temperature superplasticity, Zinc alloys

This work reports a simple and scalable strategy to prepare a series of thermoresponsive polyurethanes synthesized via copolymerization of dicyclohexyl diisocyanate with glycerol ethoxylate in a single one-pot system. These polyurethanes exhibit lower critical solution temperatures (LCST) at 57 degrees C. The LCST of synthesized polyurethane was determined from Dynamic Scanning Calorimetry and UV-vis measurements. Both the LCST and T-g of synthesized polyurethane was tuned by varying the ratio between hard segment (dicyclohexyl diisocyanate) and soft segment (glycerol ethoxylate). Thus, T-g values could be tuned from -54.6 degrees C to -19.9 degrees C for samples with different flexibility. The swelling and deswelling studies were done at room temperature and above the LCST respectively. The results showed that the swelling ratio increases with the increase of soft segment (glycerol ethoxylate) in synthesized polyurethanes. Furthermore, the mechanical properties of the membrane were studied by universal tensile testing measurements. Specifically, stress at break values varied from 0.35 +/- 0.07 MPa to 0.91 +/- 0.15 MPa for the tested membranes, whereas elongation at break data ranged from 101.9 +/- 20.9 % to 192.4 +/- 24.4 %, and Young's modulus varied from 0.35 +/- 0.03 MPa to 1.85 +/- 0.19 MPa. Tensile strength of the films increased with the increase of the hard segment and elongation at break decreased.

JTD Keywords: copolymerization, critical solution temperatures, polyurethanes, tensile strength, Biodegradable polyurethanes, Copolymerization, Critical solution temperatures, Glycol), Polymers, Polyurethanes, Solvent-free, Tensile strength, Thermoresponsive materials

Enteroaggregative Escherichia coli (EAEC) strains are one of the diarrheagenic pathotypes. EAEC strains harbor a virulence plasmid (pAA2) that encodes, among other virulence determinants, the aggR gene. The expression of the AggR protein leads to the expression of several virulence determinants in both plasmids and chromosomes. In this work, we describe a novel mechanism that influences AggR expression. Because of the absence of a Rho-independent terminator in the 3?UTR, aggR transcripts extend far beyond the aggR ORF. These transcripts are prone to PNPase-mediated degradation. Structural alterations in the 3?UTR result in increased aggR transcript stability, leading to increased AggR levels. We therefore investigated the effect of increased AggR levels on EAEC virulence. Upon finding the previously described AggR-dependent virulence factors, we detected novel AggR-regulated genes that may play relevant roles in EAEC virulence. Mutants exhibiting high AggR levels because of structural alterations in the aggR 3?UTR show increased mobility and increased pAA2 conjugation frequency. Furthermore, among the genes exhibiting increased fold change values, we could identify those of metabolic pathways that promote increased degradation of arginine, fatty acids and gamma-aminobutyric acid (GABA), respectively. In this paper, we discuss how the AggR-dependent increase in specific metabolic pathways activity may contribute to EAEC virulence.

JTD Keywords: aggregative adherence, arginine metabolism, biofilm formation, escherichia-coli, gene-expression, messenger-rna, operon, persistent diarrhea, untranslated region, Aggr protein, e coli, Escherichia coli, Escherichia coli proteins, Fimbria-i expression, Trans-activators, Virulence

Tau hyperphosphorylation is the first step of neurofibrillary tangle (NFT) formation. In the present study, samples of the entorhinal cortex (EC) and frontal cortex area 8 (FC) of cases with NFT pathology classified as stages I–II, III–IV, and V–VI without comorbidities, and of middle-aged (MA) individuals with no NFT pathology, were analyzed by conventional label-free and SWATH-MS (sequential window acquisition of all theoretical fragment ion spectra mass spectrometry) to assess the (phospho)proteomes. The total number of identified dysregulated phosphoproteins was 214 in the EC, 65 of which were dysregulated at the first stages (I–II) of NFT pathology; 167 phosphoproteins were dysregulated in the FC, 81 of them at stages I–II of NFT pathology. A large percentage of dysregulated phosphoproteins were identified in the two regions and at different stages of NFT progression. The main group of dysregulated phosphoproteins was made up of components of the membranes, cytoskeleton, synapses, proteins linked to membrane transport and ion channels, and kinases. The present results show abnormal phosphorylation of proteins at the first stages of NFT pathology in the elderly (in individuals clinically considered representative of normal aging) and sporadic Alzheimer's disease (sAD). Dysregulated protein phosphorylation in the FC precedes the formation of NFTs and SPs. The most active period of dysregulated phosphorylation is at stages III–IV when a subpopulation of individuals might be clinically categorized as suffering from mild cognitive impairment which is a preceding determinant stage in the progression to dementia. Altered phosphorylation of selected proteins, carried out by activation of several kinases, may alter membrane and cytoskeletal functions, among them synaptic transmission and membrane/cytoskeleton signaling. Besides their implications in sAD, the present observations suggest a molecular substrate for “benign” cognitive deterioration in “normal” brain aging.

JTD Keywords: (phospho)proteomics, alzheimer's disease, amyloid-beta, association guidelines, brain aging, cytoskeleton, frontal-cortex, kinases, lipid rafts, membranes, national institute, neuropathologic assessment, pathological process, protein phosphorylation, synapse pathology, synapses, tau, tau pathology, (phospho)proteomics, Adult, Age-related tauopathy, Aged, Aged, 80 and over, Aging, Alzheimer disease, Alzheimer's disease, Brain, Brain aging, Cytoskeleton, Female, Humans, Kinases, Male, Membranes, Middle aged, Neurofibrillary tangles, Phosphorylation, Protein phosphorylation, Synapses, Tau, Tau proteins

Chronic wounds represent a major health burden and drain on medical system. Efficient wound repair is only possible if the dressing materials target simultaneously multiple factors involved in wound chronicity, such as deleterious proteolytic and oxidative enzymes and high bacterial load. Here we develop multifunctional hydrogels for chronic wound management through self-assembling of thiolated hyaluronic acid (HA-SH) and bioactive silver-lignin nanoparticles (Ag@Lig NPs). Dynamic and reversible interactions between the polymer and Ag@Lig NPs yield hybrid nanocomposite hydrogels with shear-thinning and self-healing properties, coupled to zero-order kinetics release of antimicrobial silver in response to infection-related hyalurodinase. The hydrogels inhibit the major enzymes myeloperoxidase and matrix metalloproteinases responsible for wound chronicity in a patient's wound exudate. Furthermore, the lignin-capped AgNPs provide the hydrogel with antioxidant properties and strong antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa. The nanocomposite hydrogels are not toxic to human keratinocytes after 7 days of direct contact. Complete tissue remodeling and restoration of skin integrity is demonstrated in vivo in a diabetic mouse model. Hematological analysis reveals lack of wound inflammation due to bacterial infection or toxicity, confirming the potential of HA-SH/Ag@Lig NPs hydrogels for chronic wound management. Statement of significance: Multifunctional hydrogels are promising materials to promote healing of complex wounds. Herein, we report simple and versatile route to prepare biocompatible and multifunctional self-assembled hydrogels for efficient chronic wound treatment utilizing polymer-nanoparticle interactions. Hybrid silver-lignin nanoparticles (Ag@Lig NPs) played both: i) structural role, acting as crosslinking nodes in the hydrogel and endowing it with shear-thinning (ability to flow under applied shear stress) and self-healing properties, and ii) functional role, imparting strong antibacterial and antioxidant activity. Remarkably, the in situ self-assembling of thiolated hyaluronic acid and Ag@Lig NPs yields nanocomposite hydrogels able to simultaneously inhibits the major factors involved in wound chronicity, namely the overexpressed deleterious proteolytic and oxidative enzymes, and high bacterial load.

JTD Keywords: catechol, chronic wounds, dressing materials, inhibition, mechanism, nano-enabled hydrogels, polyphenols, promogran, self-assembling, silver-lignin nanoparticles, systems, tannins, Chronic wounds, Degradation, Dressing materials, Nano-enabled hydrogels, Self-assembling, Silver-lignin nanoparticles, Thiolated hyaluronic acid

The unique ability to identify one's own body and experience it as one's own is fundamental in goal-oriented behavior and survival. However, the mechanisms underlying the so-called body ownership are yet not fully understood. Evidence based on Rubber Hand Illusion (RHI) paradigms has demonstrated that body ownership is a product of reception and integration of self and externally generated multisensory information, feedforward and feedback processing of sensorimotor signals, and prior knowledge about the body. Crucially, however, these designs commonly involve the processing of proximal modalities while the contribution of distal sensory signals to the experience of ownership remains elusive. Here we propose that, like any robust percept, body ownership depends on the integration and prediction across all sensory modalities, including distal sensory signals pertaining to the environment. To test our hypothesis, we created an embodied goal-oriented Virtual Air Hockey Task, in which participants were to hit a virtual puck into a goal. In two conditions, we manipulated the congruency of distal multisensory cues (auditory and visual) while preserving proximal and action-driven signals entirely predictable. Compared to a fully congruent condition, our results revealed a significant decrease on three dimensions of ownership evaluation when distal signals were incongruent, including the subjective report as well as physiological and kinematic responses to an unexpected threat. Together, these findings support the notion that the way we represent our body is contingent upon all the sensory stimuli, including distal and action-independent signals. The present data extend the current framework of body ownership and may also find applications in rehabilitation scenarios.