Publications

PurposeBiological matrices have been used to reinforce large rotator cuff tear repairs. However, rapid resorption and initial immune reactions presented challenges in clinical practice. This study evaluates whether a resorbable synthetic matrix (scaffold), used alone or with platelet-rich plasma (PRP), impacts repair processes at microscopic, ultrasound, and biomechanical levels in a rabbit model of induced tendon-bone interface injury.MethodsAn experimental study was performed on 24 rabbits. Two experimental groups (n = 12 each) and a control group (n = 24) were defined. In the first group (BioP), the internal gastrocnemius tendon was sectioned and repaired to bone using double-row sutures, reinforced with a PLC (poly-L-lactic-co-epsilon-caprolactone) and PLA (polylactic acid) scaffold. In the second group (BioP + PRP), autologous PRP was added to the repair. The control group received no scaffold or PRP. Euthanasia was performed at 8 weeks, followed by microscopic, ultrasound, and biomechanical evaluations.ResultsMicroscopically, a granulomatous reaction limited to the foreign body was observed in both scaffold groups. The healing process was not altered in any group, showing good biocompatibility of the scaffold. Echographically, a greater sagittal diameter was observed in the group without PRP compared to the other groups. Biomechanically, no significant differences in rupture zones were found across groups, but the scaffold-only group required a higher maximum applied force before rupture.ConclusionsAt 8 weeks, using a degradable synthetic PLC and PLA scaffold as support at the bone-tendon interface did not significantly alter the normal repair process, showed echographic and biomechanical benefits, and PRP did not show additional benefits in our experimental model.

JTD Keywords: Augmentation, Biology, Biomaterials, Cells, Efficacy, Ge, Matrix, Platelet-rich plasma, Regeneration, Rotator cuff repair, Shoulder, Surgical repair, Technologies, Tissue engineerin

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

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

Lagunas, Anna

JTD

The urgent need for safer and innovative antitubercular agents remains a priority for the scientific community. In pursuit of this goal, we designed and evaluated novel 5-phenylfuran-2-carboxylic acid derivatives targeting Mycobacterium tuberculosis (Mtb) salicylate synthase (MbtI), a key enzyme, absent in humans, that plays a crucial role in Mtb virulence. Several potent MbtI inhibitors demonstrating significant antitubercular activity and a favorable safety profile were identified. Structure-guided optimization yielded 5-(3-cyano-5-isobutoxyphenyl)furan-2-carboxylic acid (1e), which exhibited strong MbtI inhibition (IC50 = 11.2 mu M) and a promising in vitro antitubercular activity (MIC99 = 32 mu M against M. bovis BCG). Esters of 1e were effectively loaded into poly(2-methacryloyloxyethyl phosphorylcholine)-poly(2-(diisopropylamino)ethyl methacrylate) (PMPC-PDPA) polymersomes (POs) and delivered to intracellular mycobacteria, resulting in reduced Mtb viability. This study provides a foundation for the use of POs in the development of future MbtI-targeted therapies for tuberculosis.

JTD Keywords: Chemistry, Discovery, Drug-delivery, Insigh, Polymersomes, Salicylate synthase mbti, Siderophore, Strategies, Target, Visualization

Odorant discrimination mechanisms are based on the differential interactions between odorant molecules and olfactory receptors (ORs). Biohybrid sensors based on ORs described to date show selectivity towards specific versus non-specific binding of odorants, being unable to distinguish between specific ligands of different affinity. Here we disclose a method that enables odorant discrimination based on the modulation of the capacitive response of the receptor, which allows the differentiation of three high-affinity hOR1A1 agonists. We performed voltammetry and impedance measurements of the hOR1A1 receptor selectively immobilized on a gold electrode in the absence and presence of the agonists. Binding induces a decrease in the capacitive response of the receptor that is proportional to the ligand potency, reaching up to a 40% decrease for the cognate ligand dihydrojasmone, which is attributed to changes in the magnitude and orientation of the electric dipole in the receptor, thereby regulating its response to the applied electric field.

JTD Keywords: Agonist, Biosenso, Electric dipol, Electrochemical impedance spectroscopy, Impedance, Nose, Odorant receptors, Olfactory receptor, Selectivity, Specific capacitance

Objectives: Despite the high survival rates of dental implants, peri-implantitis is a prevalent complication. Periimplantitis is related to biofilm that adheres to the surface of implants and causes peri-implant chronic inflammation and bone destruction. Different surface treatments have been proposed to prevent biofilm formation. The objective of this systematic review was analyzing different types of antimicrobial coatings and identifying the most effective one(s) to control bacterial colonization over extended periods of analysis. Data, sources and study selection: We performed a bibliographic search in Pubmed and Cochrane base of articles published after 2010 to answer, according to the PICO system, the following question: What is the most effective antibacterial surface coating for dental implants? Only papers including a minimum follow-up bacteria growth analysis for at least 48 h were selected. After selection, the studies were classified using the PRISMA system. A total of 40 studies were included. Conclusions: Three main categories of coatings were identified: Antibacterial peptides, synthetic antimicrobial molecules (polymers, antibiotics, ...), and metallic nanoparticles (silver). Antibacterial peptide coatings to modify dental implant surfaces have been the most studied and effective surface modification to control bacterial colonization over extended periods of incubation as they are highly potent, durable and biocompatible. However, more in vitro and pre-clinical studies are needed to assess their true potential as a technology for preventing periimplant infections.

JTD Keywords: Anti-infective coating, Antibiotics, Antimicrobial peptide coatings, Antimicrobial peptides, Antimicrobial polymers, Bacterial colonizatio, Biofilm formatio, Cationic peptides, Chimeric peptides, Dental implants, Human gingival fibroblasts, Metal nanoparticles, Osseointegrated oral implants, Peri-implantitis, Silver nanoparticles, Surface treatment, Sustained-release device, Titanium surfaces

Acetylcholine and the cholinergic system are crucial to brain function, including functions such as consciousness and cognition. Dysregulation of this system is implicated in the pathophysiology of neurological conditions such as Alzheimer's disease. For this reason, cholinergic neuromodulation is relevant in both basic neuroscience and clinical neurology. In this study, we used photopharmacology to modulate neuronal activity using the novel selective type-1 muscarinic (M1) photoswitchable drugs: the agonist benzyl quinolone carboxylic acid-azo-iperoxo (BAI) and the antagonist cryptozepine-2. Our aim was to investigate the control over these cholinergic receptors using light and to investigate the effects of these drugs on physiological spontaneous slow waves and on epileptic activity in the cerebral cortex. First, we used transfected HEK cell cultures and demonstrated BAI's preferential activation of M1 muscarinic acetylcholine receptors (mAChRs) compared with M2 mAChRs. Next, we found that white-light illumination of BAI increased the frequency of spontaneous slow-wave activity in brain cortical networks of both active slices and anesthetized mice, through M1-mAChRs activation. Illumination of cryptozepine-2 with UV light effectively suppressed not only the muscarinic-induced increase in slow-wave frequency, but also muscarinic-induced epileptiform discharges. These findings not only shed light on the role of M1 acetylcholine receptors in the cortical network dynamics but also lay the groundwork for developing advanced light-based pharmacological therapies. Photopharmacology offers the potential for high-precision spatiotemporal control of brain networks with high pharmacological specificity in both healthy and pathological conditions.

JTD Keywords: Acetylcholine, Acetylcholine-receptors, Biological health and medical sciences, Brain, Epilepsy, Hz oscillation, Less-than-1 hz, Modulation, Network mechanisms, Neuromodulation, Neuroscienc, Pathology, Photopharmacology, Seizures, Slee, Slow oscillations

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

Cell membranes play a key role in bottom-up synthetic biology, as they enable interaction control, transport, and other essential functions. These ultra-thin, flexible, yet stable structures form through the self-assembly of lipids and proteins. While liposomes are common mimics, their synthetic membranes often fail to replicate natural properties due to poor structural control. To address this, pepticombs are introduced, a new family of supramolecular building blocks. They are synthesized by regularly appending anionic surfactants with lipid-long alkyl tails to cationic amino acid residues of recombinant elastin-like supercharged unfolded polypeptides (SUPs). Using microscopy techniques and molecular dynamics simulations, the formation of giant unilamellar vesicles, termed pepticombisomes, is demonstrated and their membrane properties are characterized. The molecular topology of pepticombs allows for precise mimicry of membrane thickness and flexibility, beyond classic polymersomes. Unlike the previously introduced ionically-linked comb polymers, all pepticombs exhibit a uniform degree of polymerization, composition, sequence, and spontaneous curvature. This uniformity ensures consistent hydrophobic tail distribution, facilitating intermolecular hydrogen bonding within the backbone. This generates elastic heterogeneities and the concomitant formation of non-icosahedral faceted vesicles, as previously predicted. Additionally, pepticombisomes can incorporate functional lipids, enhancing design flexibility.

JTD Keywords: Biomimetic synthesis, Bottom-up synthetic biology, Cholesterol, Dynamics, Glycodendrimersomes, Janus dendrimers, Lipids, Nanoscal, Organization, Polymersome membranes, Protein, Stability, Supercharged peptides, Synthetic cells, Vesicle

Metabolism in biological systems involves the continuous formation and breakdown of chemical and structural components, driven by chemical energy. In specific, metabolic processes on cellular membranes result in in situ formation and degradation of the constituent phospholipid molecules, by consuming fuel, to dynamically regulate the properties. Synthetic analogs of such chemically fueled phospholipid vesicles have been challenging. Here we report a bio-inspired approach for the in situ formation of phospholipids, from water soluble precursors, and their fuel driven self-assembly into vesicles. We show that the kinetic competition between anabolic and catabolic-like reactions leads to the formation and enzymatic degradation of the double-tailed, vesicle-forming phospholipid. Spectroscopic and microscopic analysis demonstrate the formation of transient vesicles whose lifetime can be easily tuned from minutes to hours. Importantly, our design results in the formation of uniform sized (65 nm) vesicles simply by mixing the precursors, thus avoiding the traditional complex methods. Finally, our sub-100 nm vesicles are of the right size for application in drug delivery. We have demonstrated that the release kinetics of the incorporated cargo molecules can be dynamically regulated for potential applications in adaptive nanomedicine.

JTD Keywords: Droplets, Mode, Phospholipids, Supramolecular chemistry, Systems chemistry, Transient assembl, Vesicles

Flexible and biocompatible strain sensors are becoming increasingly important in fields such as health monitoring, wearable electronics, and environmental sensing because they offer significant advantages over conventional rigid systems. However, they lack the versatility and ecological and physiological biocompatibility necessary for broader integration within biological systems. Here, we describe the development of an inexpensive water-based plasticized chitosan-carbon black composite ink that can be used to produce conductive and biocompatible strain sensors. The ink can be applied to various surfaces, including skin, internal organs, and other biological tissues, using numerous methods, such as painting, dipping, and stamping. Furthermore, this unprecedented ability to attach and conform to biological surfaces allows the exploration of secondary sensing innovations, such as exploiting skin wrinkles to improve sensitivity. This study demonstrates that the ink exhibits a reliable change in electrical resistance in response to a wide range of motions, from subtle vibrations during speech and heartbeats to extensive articulations, like finger and elbow movements. This exceptional sensitivity range, biocompatibility, and the ink's low cost, biodegradability, and ease of removal enhance its applicability in sustainable, temporary, and customizable sensing solutions, highlighting its potential for versatile applications in human health monitoring, motion detection, and environmental sensing.

JTD Keywords: Blac, Chitin, Composites, Performance strain sensors

The Boston keratoprosthesis (BKPro) is a critical device for vision restoration in complex cases of corneal blindness, although its long-term retention is challenged by infection risks. This study aims to enhance the antimicrobial properties of the titanium (Ti) backplate of the BKPro by ion implanting silver and copper ions to achieve effective infection control while maintaining cytocompatibility. Research on antimicrobial modifications for BKPro is limited, and while metallic ions like Ag and Cu show promise for biomaterial improvement, their effects on human corneal keratocytes (HCKs) require further study. Ag and Cu were implanted onto rough Ti surfaces, as mono- and coimplantations. Cytotoxicity was assessed in HCKs, and antimicrobial efficacy was tested against Pseudomonas aeruginosa and Candida albicans. After 21 d, monoimplanted Ag samples released 300.4 ppb of Ag+, coimplanted samples released 427.5 ppb of Ag+ and 272.3 ppb of Cu ions, and monoimplanted Cu samples released 567.0 ppb of Cu ions. All ion-implanted surfaces supported HCK proliferation, exhibited no cytotoxicity, and showed strong antimicrobial activity. Ag-implanted surfaces provided antibacterial effects through membrane disruption and reactive oxygen species generation, while Cu-implanted surfaces exhibited antifungal effects via impaired enzymatic functions and reactive oxygen species. Coimplanted AgCu surfaces demonstrated synergistic antimicrobial effects, resulting from the synergy between the bactericidal actions of Ag and the oxidative stress contributions of Cu. Additionally, ion-implanted surfaces enhanced HCK adhesion under co-culture conditions. In conclusion, ion implantation effectively imparts antimicrobial properties to the Ti backplate of BKPro, reducing infection risks while preserving compatibility with corneal cells.

JTD Keywords: Endophthalmitis, Mechanisms, Nanoparticle, Silver

In this study, the extraction process of grape pomace from the Lebanese autochthonous cultivar Asswad Karech was enhanced through the selection of specific parameters, yielding an antioxidant extract (20 mg/mL) that was co-loaded with resveratrol (5 mg/mL) into phospholipid vesicles containing penetration enhancers (PEVs). Propylene glycol (PG) was incorporated as a penetration enhancer at concentrations of 10, 20, and 30 % to obtain 10 PG-PEVs, 20 PG-PEVs, and 30 PG-PEVs. Vesicle preparation was achieved through direct sonication, yielding unilamellar and bilamellar vesicles with an average size of similar to 205; 234 nm, a monodisperse distribution (polydispersity index

JTD Keywords: By-product valorisation, Cigarette smoke, Delivery, Dru, Grape pomace extract, In-vitro, Liposomes, Lung deliver, Oxidative stress, Phospholipids vesicles

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: Confinement, Dynamics, Epithelial cells, Extracellular matrix, Extracellular-matrix, Force transmission, Hydrogels, Mechanics, Micropatterning, Migration, Morphology, Motilit, Polyacrylamide hydrogels, Stiffness, Substrate, Viscoelasticit, Viscoelasticity

The chemical and physical stability of bio-hydrogels are of utmost interest to avoid the premature degradation of the polymer and to favor cyclic material operations (i.e., material recovery and re-using). In this work, the stability of different alginate hydrogels semi-interpenetrated with poly(3,4-ethylenedioxythiophene):polystyrene sulfonate conducting polymer (Alg/PEDOT), which acts as a photothermal absorber is examined. More specifically, the behavior of Alg/PEDOT hydrogels ionically and covalently crosslinked with Ca2+ ions and glyoxal, respectively, has been compared when used as water purification platforms. The homogenous porosity and higher cycling capacity of the glyoxal-crosslinked gels provide superior performance for water-steam generation under sunlight irradiation than that of the ionically stabilized gel. Furthermore, increasing the glyoxal cross-linking reaction time prove to have little effect on the porosity and the efficiency of freshwater supply from an artificial seawater solution. Covalent cross-links provide thermal absorber (PEDOT:PSS) retention capacity in artificial seawater, which is critical to maintaining such efficiency with the increasing number of purification cycles. This research opens new frontiers to promote the use of alginate biopolymer in chemical engineering processes such as water desalination, directly addressing the United Nations Sustainable Development Goals for Clean Water & Life on Land.

JTD Keywords: 4-ethylenedioxythiophene, Alginate polysaccharide, Cell delivery, Glyoxal, Interpenetrating hydrogel network, Poly(3, Raman-spectroscopy, Sodium alginate, Tissu

In serology, each sample is typically tested individually, one antigen at a time. This is costly and time consuming. Serology techniques should ideally allow recurrent measurements in parallel in small sample volumes and be inexpensive and fast. Here we show that mass cytometry can be used to scale up multiplexed serology testing by leveraging polystyrene beads uniformly loaded with combinations of stable isotopes. We generated 18,480 unique isotopically barcoded beads to simultaneously detect, in a single tube with 924 serum samples, the levels of immunoglobulins G and M against 19 proteins from SARS-CoV-2 (a total of 36,960 tests in 400 nl of sample volume and 30 mu l of reaction volume). As a rapid, high-throughput and cost-effective technique, serology by mass cytometry may contribute to the effective management of public health emergencies originating from infectious diseases.

JTD Keywords: Biolog, Cytometer, Transmission

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

Defining the trajectory of cells during differentiation and disease is key for uncovering the mechanisms driving cell fate and identity. However, trajectories of human cells remain largely unexplored due to the challenges of studying them with human samples. In this study, we investigate the proteome trajectory of iPSCs differentiation to hepatic stellate cells (diHSCs) and identify RORA as a key transcription factor governing the metabolic reprogramming of HSCs necessary for diHSCs' commitment, identity, and activation. Using RORA deficient iPSCs and pharmacologic interventions, we show that RORA is required for early differentiation and prevents diHSCs activation by reducing the high energetic state of the cells. While RORA knockout mice have enhanced fibrosis, RORA agonists rescue multi-organ fibrosis in in vivo models. Notably, RORA expression correlates negatively with liver fibrosis and HSCs activation markers in patients with liver disease. This study reveals that RORA regulates cell metabolic plasticity, important for mesoderm differentiation, pericyte quiescence, and fibrosis, influencing cell commitment and disease.

JTD Keywords: Generation, Inhibition, Mic

Historical processes in community assembly, such as species arrival order, influence interactions, causing priority effects. Candida albicans and Pseudomonas aeruginosa often co-occur in biofilm-based infections of the skin, lungs, and medical devices. Their predominantly antagonistic relationship involves complex physical and chemical interactions. However, the presence and implications of priority effects among these microorganisms remain largely unexplored. Here, we investigate the presence and impact of priority effect in dual-species biofilms using clinical isolates. By varying inoculation order, we observe significant changes in biofilm composition, structure, virulence, and antimicrobial susceptibility. The first colonizer has an advantage for surface colonization. Consecutive colonization increases biofilm virulence and negates C. albicans' protective effect on P. aeruginosa PAET1 against meropenem treatment. Finally, we propose N-acetylcysteine as an adjuvant for treating C. albicans and P. aeruginosa interkingdom infections, working independently of priority effects.

JTD Keywords: Airway colonization, Bacteria, Infections, Pneumonia, Spp., Virulenc

Immunotherapies are beneficial for a considerable proportion of cancer patients, but ineffective in others. In vitro modelling of the complex interactions between cancer cells and their microenvironment could provide a path to understanding immune therapy sensitivity and resistance. Here we develop MIRO, a fully humanised in vitro platform to model the spatial organisation of the tumour/stroma interface and its interaction with immune cells. We find that stromal barriers are associated with immune exclusion and protect cancer cells from antibody-dependent cellular cytotoxicity, elicited by targeted therapy. We demonstrate that IL2-driven immunomodulation increases immune cell velocity and spreading to overcome stromal immunosuppression and restores anti-cancer response in refractory tumours. Collectively, our study underscores the translational value of MIRO as a powerful tool for exploring how the spatial organisation of the tumour microenvironment shapes the immune landscape and influences the responses to immunomodulating therapies.

JTD Keywords: Activation, Animals, Architecture, Breast-cancer, Cancer-associated fibroblasts, Cell line, tumor, Collagen, Female, Humans, Immunomodulation, Immunotherapy, Interleukin-2, Lab-on-a-chip devices, Mechanism, Mice, Microenvironment, Migration, Neoplasms, Stromal cells, T-cells, Therap, Tumor microenvironment

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

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

ObjectivesThis study evaluated different designs of the conical implant-abutment connection (IAC) and their resistance to microgap formation under oblique loads as specified by the ISO standard for testing dental implants. Also evaluated was the effect of deviations from the ISO specifications on the outcomes.MethodsFinite element analysis was conducted to compare the microgap formation and stress distribution among three conical IAC designs (A, B, and C) in two loading configurations: one compliant with ISO 14801 and one with a modified load adaptor (non-ISO). The different IAC designs varied in the taper, diameter, and cone height. The cone angle mismatch (Cam) between the implant and abutment was considered. A torque of 20 Ncm and oblique loads (up to 400 N) were simulated.ResultsThe stresses produced by the screw-tightening torque varied among the different IAC designs. The contact height was approximately 0.3 mm for Designs A and B, and less than 0.03 mm for Design C. Under oblique loads, Design A maintained IAC sealing without gap formation up to 400 N. With the ISO adaptor, gaps appeared in Design B at 300 N and in Design C at 90 N. The non-ISO adaptor resulted in gap formation at 160 N for Design B and at 50 N for Design C.ConclusionsThe IAC design and cone angle mismatch significantly influenced microgap formation, with some designs showing zero gaps even when the oblique load reached 400 N. The non-ISO adaptor increased gap formation in IACs B and C.

JTD Keywords: Bacterial leakage, Behavior, Dental implant, Dental implant-abutments design, Dimensional measurement accuracy, Finite element analysis, In-vitro, Interface, Mechanical, Peri-implantitis, Scre, Sealant agents, Stres, Taper

Dementia affects a large proportion of the world's population. Approaches that allow for early disease detection and non-invasive monitoring of disease progression are desperately needed. Current approaches are centred on costly imaging technologies such as positron emission tomography and magnetic resonance imaging. We propose an alternative approach to assess neurodegeneration based on diffuse correlation spectroscopy (DCS), a remote and optical sensing technique. We employ this approach to assess neurodegeneration in mouse brains from healthy animals and those with prion disease. We find a statistically significant difference in the optical speckle decor relation times between prion-diseased and healthy animals. We directly calibrated our DCS technique using hydrogel samples of varying Young's modulus, indicating that we can optically measure changes in the brain tissue stiffness in the order of 60 Pa (corresponding to a 1 s change in speckle decor relation time). DCS holds promise for contact-free assessment of tissue stiffness alteration due to neurodegeneration, with a similar sensitivity to contact-based (e.g. nanoindentation) approaches.

JTD Keywords: Correlation spectroscopy, Decorrelation time, Disease, In-vivo, Magnetic-resonance elastography, Prion neurodegeneration, Tomograph

Gas sensor-based electronic noses (e-noses) have gained considerable attention over the past thirty years, leading to the publication of numerous research studies focused on both the development of these instruments and their various applications. Nonetheless, the limited specificity of gas sensors, along with the common requirement for chemical identification, has led to the adaptation and incorporation of analytical chemistry instruments into the e-nose framework. Although instrument-based e-noses exhibit greater specificity to gasses than traditional ones, they still produce data that require correction in order to build reliable predictive models. In this work, we introduce the use of a multivariate signal processing workflow for datasets from a multi-capillary column ion mobility spectrometer-based e-nose. Adhering to the electronic nose philosophy, these workflows prioritized untargeted approaches, avoiding dependence on traditional peak integration techniques. A comprehensive validation process demonstrates that the application of this preprocessing strategy not only mitigates overfitting but also produces parsimonious models, where classification accuracy is maintained with simpler, more interpretable structures. This reduction in model complexity offers significant advantages, providing more efficient and robust models without compromising predictive performance. This strategy was successfully tested on an olive oil dataset, showcasing its capability to improve model parsimony and generalization performance.

JTD Keywords: Breath, Chromatography, Classification, Electronic nose, Hs-gc-ims, Ion mobility spectrometry, Mcc-ims-based e-nose, Olive oi, Olive oil, Parsimony, Preprocessing, Quality control, Selection, Signal processing workflow, System, Too, Validation, Wavelet

The intricate interactions between the host immune system and its microbiome constituents undergo dynamic shifts in response to perturbations to the intestinal tissue environment. Our ability to study these events on the systems level is significantly limited by in situ approaches capable of generating simultaneous insights from both host and microbial communities. Here, we introduce Microbiome Cartography (MicroCart), a framework for simultaneous in situ probing of host and microbiome across multiple spatial modalities. We demonstrate MicroCart by investigating gut host and microbiome changes in a murine colitis model, using spatial proteomics, transcriptomics, and glycomics. Our findings reveal a global but systematic transformation in tissue immune responses, encompassing tissue-level remodeling in response to host immune and epithelial cell state perturbations, bacterial population shifts, localized inflammatory responses, and metabolic process alterations during colitis. MicroCart enables a deep investigation of the intricate interplay between the host tissue and its microbiome with spatial multi-omics.

JTD Keywords: Animals, Bacteria, Cellular microenvironment, Colitis, Design, Disease models, animal, Environment, Fis, Gastrointestinal microbiome, Glycomics, Host microbial interactions, Intestinal mucosa, Intestines, Mice, Mice, inbred c57bl, Multiomics, Organization, Probes, Proteins, Proteomics, Rna, Subcellular resolution, Transcriptome

Nanoparticle (NP) morphology holds significant importance in nanomedicine, particularly concerning its implications for biological responses. This study investigates the impact of synthesizing polymers with varying degrees of methionine (MET) polymerization on three distinct drug delivery systems: spherical micelles, worm-like micelles, and vesicles, all loaded with the photosensitizer chlorin e6 (Ce6). We analyzed their distribution at both cellular and animal levels, revealing how NP morphology influences cellular uptake, subcellular localization, penetration of multicellular spheroids, blood half-life, and biodistributions across major organs. Employing a physiologically based pharmacokinetic (PBPK) model enabled us to simulate diverse distribution patterns and quantify the targeting efficiency of NPs toward tumors. Our investigation elucidates that spherical micelles exhibit lower accumulation levels within the reticuloendothelial system, potentially mitigating adverse side effects despite their higher glomerular filtration rate. This nuanced understanding underscores the complex interplay between NP morphology and biological responses, providing valuable insights into optimizing therapeutic efficacy while minimizing undesirable effects. We thus report the integration of experimental analyses with PBPK modeling to elucidate the topological characteristics of NP, thereby shedding light on their distribution patterns, therapeutic efficacy, and potential side effects.

JTD Keywords: Drug, Nanorod, Polymersomes, Strategies

The global dental implant market is projected to reach $9.5 billion by 2032, growing at a 6.5% compound annual growth rate due to the rising prevalence of dental diseases. Importantly, this growth raises concerns about postoperative infections, which present significant challenges within our healthcare system and lead to a two-thirds failure rate for infected implants. In this study, we present an innovative multilevel coating system that makes the surface of dental titanium implants resistant to bacterial colonization, thereby minimizing the risk of infection development. This multilevel coating features a nanometer-thick biohybrid coating layer combined with a microgroove surface microstructuring, creating physical barriers that enhance the stability of the biohybrids against mechanical abrasion. Our coating demonstrates excellent biocompatibility and strong antifouling properties against undiluted blood plasma proteins. Furthermore, the combination of surface microstructuring and the biohybrid coating remains stable under prolonged mechanical stress simulation and effectively repels clinically relevant bacteria, achieving a 99% reduction in bacterial colonization on the implant. These findings underscore the potential of this approach to prevent implant-associated infections and highlight the critical role of surface engineering in ensuring long-term implant performance.

JTD Keywords: Antifouling, Bacteriarepellency, Biofilm formatio, Dental titanium implants, Infection preventio, Protein-polymer biohybrids, Ti-binding peptide, Usp laser microstructuring

We recently characterized the potent antiplasmodial activity of the aggregated protein dye YAT2150, whose presumed mode of action is the inhibition of protein aggregation in the malaria parasite. Using single-dose and ramping methods, assays were done to select Plasmodium falciparum parasites resistant to YAT2150 concentrations ranging from 3x to 0.25x the in vitro IC50 of the compound (in the two-digit nM range) and performed a cross-resistance assessment in P. falciparum lines harboring mutations that make them resistant to a variety of antimalarial drugs. Resistant parasites did not emerge in vitro after 60 days of incubation, which postulates YAT2150 as an 'irresistible' antimalarial. The lyophilized compound is stable for at least one year stored at 25 degrees C. Tests performed in clinical isolates indicated that YAT2150 had also strong activity against Plasmodium vivax (IC50 between 4 and 36 nM) and Leishmania infantum (1.27 and 1.11 mu M), placing it as a unique compound with perspectives of becoming the first drug to be used against both malaria and leishmaniasis.

JTD Keywords: Artemisinin, Complexit, Cross-resistance, Genome, In-vitro, Malaria, Mefloquine, Pfmdr1 gene, Selection

Mechanical forces influence mitochondrial dynamics through previously unexplored mechanisms. A new study demonstrates that actomyosin tension inhibits mitochondrial fission by phosphorylating a key component of the fission complex and that this event regulates the nuclear accumulation of critical transcription factors.

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With the aim of healing challenging skin wounds, electro-responsive click-hydrogels made of hyaluronic acid (clickHA) crosslinked with a modified polyethylene glycol precursor (PEG) were prepared by semi- interpenetrating a conducting polymer, poly(hydroxymethyl-3,4-ethylenedioxythiophene) (PEDOT-MeOH) by oxidative polymerization. The porosity and pore size of the mixed hydrogel, clickHA/PEDOT-MeOH, were both higher than those determined for the hydrogel without PEDOT-MeOH, while a honeycomb-like morphology with PEDOT-MeOH covering the pore walls was observed. Although such PEDOT-MeOH-induced changes did not influence the water absorption capacity of clickHA, they drastically affected the mechanical and electrochemical behavior. More specifically, the semi-interpenetration of PEDOT-MeOH into clickHA resulted in an increase of the Young's modulus, the compressive strength and, especially, the electrochemical activity. The biocompatibility and the potential for skin regeneration of clickHA/PEDOT-MeOH were preliminary assessed using viability and wound-healing assays with epithelial cells. Not only is the conducting hydrogel formulation biocompatible, but also promotes efficient cell migration by electrostimulation using a small voltage (0.5 V) for a short time (15 min). Thus, in just 1 h the wound gap was repaired, and a homogeneous monolayer of migrated cells was formed.

JTD Keywords: 4-ethylenedioxythiophene, Car, Click hydrogel, Conducting polymer, Hyaluronic acid, Poly(3, Proliferation, Rational design, Scaffold, Skin, Wound dressing

Nuclear magnetic resonance (NMR) spectroscopy is a valuable diagnostic tool limited by low sensitivity due to low nuclear spin polarization. Hyperpolarization techniques, such as dissolution dynamic nuclear polarization, significantly enhance sensitivity, enabling real-time tracking of cellular metabolism. However, traditional high-field NMR systems and bioreactor platforms pose challenges, including the need for specialized equipment and fixed sample volumes. This study introduces a scalable, 3D-printed bioreactor platform compatible with low-field NMR spectrometers, designed to accommodate bioengineered 3D cell models. The bioreactor is fabricated using biocompatible materials and features a microfluidic system for media recirculation, ensuring optimal culture conditions during NMR acquisition and cell maintenance. We characterized the NMR compatibility of the bioreactor components and confirmed minimal signal distortion. The bioreactor's efficacy was validated using HeLa and HepG2 cells, demonstrating prolonged cell viability and enhanced metabolic activity in 3D cultures compared to 2D cultures. Hyperpolarized [1-13C] pyruvate experiments revealed distinct metabolic profiles for the two cell types, highlighting the bioreactor's ability to discern metabolic profiles among samples. Our results indicate that the bioreactor platform supports the maintenance and analysis of 3D cell models in NMR studies, offering a versatile and accessible tool for metabolic and biochemical research in tissue engineering. This platform bridges the gap between advanced cellular models and NMR spectroscopy, providing a robust framework for future applications in nonspecialized laboratories. The design files for the 3D printed components are shared within the text for easy download and customization, promoting their use and adaptation for further applications.

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Chronic obstructive pulmonary disease (COPD) is a leading cause of death worldwide and greatly reduces the quality of life. Utilizing remote monitoring has been shown to improve quality of life and reduce exacerbations, but remains an ongoing area of research. We introduce a novel method for estimating changes in ease of breathing for COPD patients, using obstructed breathing data collected via wearables. Physiological signals were recorded, including respiratory airflow, acceleration, audio, and bio-impedance. By comparing patient-specific measurements, this approach enables non-intrusive remote monitoring. We analyze the influence of signal selection, window parameters, feature engineering, and classification models on predictive performance, finding that acceleration signals are most effective, complemented by audio signals. The best model achieves an F1-score of 0.83. To facilitate clinical adoption, we incorporate interpretability by designing novel saliency map methods, highlighting important aspects of the signals. We adapt local explainability techniques to time series and introduce a novel imputation method for periodic signals, improving faithfulness to the data and interpretability.

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Blood-contacting medical devices, especially extracorporeal membrane oxygenators (ECMOs), are highly susceptible to surface-induced coagulation because of their extensive surface area. This can compromise device functionality and lead to life-threatening complications. High doses of anticoagulants, combined with anti-thrombogenic surface coatings, are typically employed to mitigate this risk, but such treatment can lead to hemorrhagic complications. Therefore, bioactive surface coatings that mimic endothelial blood regulation are needed. However, evaluating these coatings under realistic ECMO conditions is both expensive and challenging. This study utilizes microchannel devices to simulate ECMO fluid dynamics and assess the clot-lysis efficacy of a self-activating fibrinolytic coating system. The system uses antifouling polymer brushes combined with tissue plasminogen activator (tPA) to induce fibrinolysis at the surface. Here, tPA catalyzes the conversion of blood plasminogen into plasmin, which dissolves clots. This positive feedback loop enhances clot digestion under ECMO-like conditions. This findings demonstrate that this coating system can significantly improve the hemocompatibility of medical device surfaces.

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Bio-nano interactions have been extensively explored in nanomedicine to develop selective delivery strategies and reduce systemic toxicity. To enhance the delivery of nanocarriers to cancer cells and improve the therapeutic efficiency, different nanomaterials have been developed. However, the limited clinical translation of nanoparticle-based therapies, largely due to issues associated with poor targeting, requires a deeper understanding of the biological phenomena underlying cell-nanoparticle interactions. In this context, we investigate the molecular and cellular mechanobiology parameters that control such interactions. We demonstrate that the pharmacological inhibition or the genetic ablation of the key mechanosensitive component of the Hippo pathway, i.e., yes-associated protein, enhances nanoparticle internalization by 1.5-fold. Importantly, this phenomenon occurs independently of nanoparticle properties, such as size, or cell properties such as surface area and stiffness. Our study reveals that the internalization of nanoparticles in target cells can be controlled by modulating cell mechanosensing pathways, potentially enhancing nanotherapy specificity.

JTD Keywords: Bio-nano interactions, Comple, Mechanobiology, Mechanotransductio, Nanoparticles, Yap/taz

The sustainable synthesis of urea from ammonia (NH3) and carbon dioxide (CO2) using ultraporous permanently polarized hydroxyapatite (upp-HAp) as catalyst has been explored as an advantageous CO2-revalorization strategy. As the simultaneous activation of N-2 and CO2 (single-step) demands an increase of the reaction conditions, we have re-visited the industrial two-step Bazarov reaction. upp-HAp has been designed as a stable multifunctional catalyst capable of promoting both CO2 and NH3 adsorption for their subsequent C-N bond formation. Herein we report the synthesis of 1 mmol/g(cat) of urea with a selectivity of 97 % under strictly mild conditions (95-120 degrees C and 1 bar of CO2; without applying any electrical currents or UV irradiation) which represents an efficiency of similar to 2 % and similar to 30 % with respect to the NH3 and CO2 content, respectively. The study of the NH3 content, products adsorbed in the catalyst, presence of intermediates and temperature of the reaction allows unveiling the great potential of upp-HAp as a green catalyst for sustainable Bazarov reactions. Results suggest that the double-step approach could be more advantageous for both synthesizing urea and as a CO2-revalorization strategy, which in turn promotes the development of specific technologies for the independent synthesis of green NH3.

JTD Keywords: Co2-revalorization, Hydroxyapatite, Permanently polarized materials, Tsp treatment, Ure, Urea

The tumor microenvironment (TME) comprises a heterogenous cell population within a complex threedimensional (3D) extracellular matrix (ECM). Stromal cells within this TME are altered by signaling cues from cancer cells to support uncontrolled tumor growth and invasion events. Moreover, the ECM also plays a fundamental role in tumor development through pathological remodeling, stiffening and interaction with TME cells. In healthy tissues, Hippo signaling pathway actively contributes to tissue growth, cell proliferation and apoptosis. However, in cancer, the Hippo signaling pathway is highly dysregulated, leading to nuclear translocation of the YAP/TAZ complex, which directly contributes to uncontrolled cell proliferation and tissue growth, and ECM remodeling and stiffening processes. Here, we compare the effect of increasing cell culture substrate stiffness, derived from tumor progression, upon the dysregulation of the Hippo signaling pathway in colorectal cancer-associated fibroblasts (CAFs) and normal colorectal fibroblasts (NFs). We correlate the dysregulation of Hippo pathway with the magnitude of the traction forces exerted by healthy and malignant stromal cells. We found that ECM stiffening is crucial in Hippo pathway dysregulation in CAFs, but not in normal fibroblasts.

JTD Keywords: Cancer-associated fibroblasts, Hippo pathway, Organ size control, Tissu, Tumor microenvironment, Yap-ta, Yap/taz

The inclusion of microexons by alternative splicing occurs frequently in neuronal proteins. The roles of these sequences are largely unknown, and changes in their degree of inclusion are associated with neurodevelopmental disorders1. We have previously shown that decreased inclusion of a 24-nucleotide neuron-specific microexon in CPEB4, a RNA-binding protein that regulates translation through cytoplasmic changes in poly(A) tail length, is linked to idiopathic autism spectrum disorder (ASD)2. Why this microexon is required and how small changes in its degree of inclusion have a dominant-negative effect on the expression of ASD-linked genes is unclear. Here we show that neuronal CPEB4 forms condensates that dissolve after depolarization, a transition associated with a switch from translational repression to activation. Heterotypic interactions between the microexon and a cluster of histidine residues prevent the irreversible aggregation of CPEB4 by competing with homotypic interactions between histidine clusters. We conclude that the microexon is required in neuronal CPEB4 to preserve the reversible regulation of CPEB4-mediated gene expression in response to neuronal stimulation.

JTD Keywords: Alternative splicing, Animals, Autism spectrum disorder, Cpeb4 protein, human, Cpeb4 protein, mouse, Exons, Gene expression regulation, Humans, Mice, Neurons, Protein aggregates, Protein biosynthesis, Rna-binding proteins