DONATE

Publications

by Keyword: PLA

Moussa, Dina G., Sharma, Ashok K., Mansour, Tamer A, Witthuhn, Bruce, Perdigão, Jorge, Rudney, Joel D., Aparicio, Conrado, Gomez, Andres, (2022). Functional signatures of ex-vivo dental caries onset Journal Of Oral Microbiology 14, 2123624

Fischer NG, Aparicio C, (2022). Junctional epithelium and hemidesmosomes: Tape and rivets for solving the “percutaneous device dilemma” in dental and other permanent implants Bioactive Materials 18, 178-198

The percutaneous device dilemma describes etiological factors, centered around the disrupted epithelial tissue surrounding non-remodelable devices, that contribute to rampant percutaneous device infection. Natural percutaneous organs, in particular their extracellular matrix mediating the “device”/epithelium interface, serve as exquisite examples to inspire longer lasting long-term percutaneous device design. For example, the tooth's imperviousness to infection is mediated by the epithelium directly surrounding it, the junctional epithelium (JE). The hallmark feature of JE is formation of hemidesmosomes, cell/matrix adhesive structures that attach surrounding oral gingiva to the tooth's enamel through a basement membrane. Here, the authors survey the multifaceted functions of the JE, emphasizing the role of the matrix, with a particular focus on hemidesmosomes and their five main components. The authors highlight the known (and unknown) effects dental implant – as a model percutaneous device – placement has on JE regeneration and synthesize this information for application to other percutaneous devices. The authors conclude with a summary of bioengineering strategies aimed at solving the percutaneous device dilemma and invigorating greater collaboration between clinicians, bioengineers, and matrix biologists. © 2022 The Authors

JTD Keywords: amino-acid-sequence, bioinspired surfaces, cell-secreted protein, growth-factor receptor, hemidesmosome, integrin beta-4 subunit, junctional epithelium, keratinocyte-derived chemokine, laminin-binding integrins, marginal bone loss, percutaneous implant, pressure wound therapy, soft-tissue integration, Bioinspired surfaces, Bullous-pemphigoid antigen, Hemidesmosome, Junctional epithelium, Percutaneous device, Percutaneous implant


Mughal, S, Lopez-Munoz, GA, Fernandez-Costa, JM, Cortes-Resendiz, A, De Chiara, F, Ramon-Azcon, J, (2022). Organs-on-Chips: Trends and Challenges in Advanced Systems Integration Advanced Materials Interfaces , 2201618

Organ-on-chip platforms combined with high-throughput sensing technology allow bridging gaps in information presented by 2D cultures modeled on static microphysiological systems. While these platforms do not aim to replicate whole organ systems with all physiological nuances, they try to mimic relevant structural, physiological, and functional features of organoids and tissues to best model disease and/or healthy states. The advent of this platform has not only challenged animal testing but has also presented the opportunity to acquire real-time, high-throughput data about the pathophysiology of disease progression by employing biosensors. Biosensors allow monitoring of the release of relevant biomarkers and metabolites as a result of physicochemical stress. It, therefore, helps conduct quick lead validation to achieve personalized medicine objectives. The organ-on-chip industry is currently embarking on an exponential growth trajectory. Multiple pharmaceutical and biotechnology companies are adopting this technology to enable quick patient-specific data acquisition at substantially low costs.

JTD Keywords: A-chip, Biosensor, Biosensors, Cancer, Cells, Culture, Disease models, Epithelial electrical-resistance, Hydrogel, Microfabrication, Microphysiological systems, Models, Niches, Organ-on-a-chips, Platform


Bouzon-Arnaiz, I, Avalos-Padilla, Y, Biosca, A, Cano-Prades, O, Roman-Alamo, L, Valle, J, Andreu, D, Moita, D, Prudencio, M, Arce, EM, Munoz-Torrero, D, Fernandez-Busquets, X, (2022). The protein aggregation inhibitor YAT2150 has potent antimalarial activity in Plasmodium falciparum in vitro cultures Bmc Biology 20, 197

Background By 2016, signs of emergence of Plasmodium falciparum resistance to artemisinin and partner drugs were detected in the Greater Mekong Subregion. Recently, the independent evolution of artemisinin resistance has also been reported in Africa and South America. This alarming scenario calls for the urgent development of new antimalarials with novel modes of action. We investigated the interference with protein aggregation, which is potentially toxic for the cell and occurs abundantly in all Plasmodium stages, as a hitherto unexplored drug target in the pathogen. Results Attempts to exacerbate the P. falciparum proteome's propensity to aggregation by delivering endogenous aggregative peptides to in vitro cultures of this parasite did not significantly affect their growth. In contrast, protein aggregation inhibitors clearly reduced the pathogen's viability. One such compound, the bis(styrylpyridinium) salt YAT2150, exhibited potent antiplasmodial activity with an in vitro IC50 of 90 nM for chloroquine- and artemisinin-resistant lines, arresting asexual blood parasites at the trophozoite stage, as well as interfering with the development of both sexual and hepatic forms of Plasmodium. At its IC50, this compound is a powerful inhibitor of the aggregation of the model amyloid beta peptide fragment 1-40, and it reduces the amount of aggregated proteins in P. falciparum cultures, suggesting that the underlying antimalarial mechanism consists in a generalized impairment of proteostasis in the pathogen. YAT2150 has an easy, rapid, and inexpensive synthesis, and because it fluoresces when it accumulates in its main localization in the Plasmodium cytosol, it is a theranostic agent. Conclusions Inhibiting protein aggregation in Plasmodium significantly reduces the parasite's viability in vitro. Since YAT2150 belongs to a novel structural class of antiplasmodials with a mode of action that potentially targets multiple gene products, rapid evolution of resistance to this drug is unlikely to occur, making it a promising compound for the post-artemisinin era.

JTD Keywords: Amyloid formation, Amyloid pan-inhibitors, Antimalarial drugs, Colocalization, Cytosolic delivery, Derivatives, Disease, Drug, In-vitro, Malaria, Mechanism, Plasmodium falciparum, Polyglutamine, Protein aggregation, Yat2150


Sole-Marti, X, Labay, C, Raymond, Y, Franch, J, Benitez, R, Ginebra, MP, Canal, C, (2022). Ceramic-hydrogel composite as carrier for cold-plasma reactive-species: Safety and osteogenic capacity in vivo Plasma Processes And Polymers , e2200155

Plasma-treated hydrogels have been put forward as a potential selective osteosarcoma therapy through the release of reactive species to the diseased site. To allow their translation to the clinics, it is crucial to show that the oxidative stress delivered by such hydrogels does not adversely affect healthy tissues. This is evaluated here by investigating the in vivo performance of a robocasted calcium phosphate cement infiltrated by a plasma-treated hydrogel. The plasma-treated composite implanted in a critical size bone defect of healthy rabbits revealed its safety, allowing equivalent bone ingrowth compared to the control scaffolds and to that of direct plasma treatment of the bone defect. This opens the door for using composite biomaterials containing plasma-generated reactive species in bone therapies.

JTD Keywords: Atmospheric plasma, Bone, Bone graft, Ceramic-hydrogel composite, Cold atmospheric plasma, Local therapy, Osteosarcoma, Plasma-treated polymer solutions, Substitutes, Survival


Fernández-Garibay, Xiomara, Gomez-Florit, Manuel, Domingues, Rui M A, Gomes, Manuela, Fernandez-Costa, Juan M., Ramon, Javier, (2022). Xeno-free bioengineered human skeletal muscle tissue using human platelet lysate-based hydrogels Biofabrication 14, 045015

Abstract Bioengineered human skeletal muscle tissues have emerged in the last years as new in vitro systems for disease modeling. These bioartificial muscles are classically fabricated by encapsulating human myogenic precursor cells in a hydrogel scaffold that resembles the extracellular matrix. However, most of these hydrogels are derived from xenogenic sources, and the culture media is supplemented with animal serum, which could interfere in drug testing assays. On the contrary, xeno-free biomaterials and culture conditions in tissue engineering offer increased relevance for developing human disease models. In this work, we used human platelet lysate-based nanocomposite hydrogels (HUgel) as scaffolds for human skeletal muscle tissue engineering. These hydrogels consist of human platelet lysate reinforced with cellulose nanocrystals (a-CNC) that allow tunable mechanical, structural, and biochemical properties for the 3D culture of stem cells. Here, we developed hydrogel casting platforms to encapsulate human muscle satellite stem cells in HUgel. The a-CNC content was modulated to enhance matrix remodeling, uniaxial tension, and self-organization of the cells, resulting in the formation of highly aligned, long myotubes expressing sarcomeric proteins. Moreover, the bioengineered human muscles were subjected to electrical stimulation, and the exerted contractile forces were measured in a non-invasive manner. Overall, our results demonstrated that the bioengineered human skeletal muscles could be built in xeno-free cell culture platforms to assess tissue functionality, which is promising for drug development applications.

JTD Keywords: 3d culture, generation, identification, image, manipulate, matrigel, mechanics, model, platelet lysate, scaffolds, tissue engineering, xeno-free, Platform, Skeletal muscle


Fontana-Escartin, A, Lanzalaco, S, Bertran, O, Aleman, C, (2022). Electrochemical multi-sensors obtained by applying an electric discharge treatment to 3D-printed poly(lactic acid) Applied Surface Science 597, 153623

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


Phuyal, Santosh, Djaerff, Elena, Le Roux, Anabel‐Lise, Baker, Martin J, Fankhauser, Daniela, Mahdizadeh, Sayyed Jalil, Reiterer, Veronika, Parizadeh, Amirabbas, Felder, Edward, Kahlhofer, Jennifer C, Teis, David, Kazanietz, Marcelo G, Geley, Stephan, Eriksson, Leif, Roca‐Cusachs, Pere, Farhan, Hesso, (2022). Mechanical strain stimulates COPII-dependent secretory trafficking via Rac1 Embo Journal 41, e110596

Martínez-Ara G, Taberner N, Takayama M, Sandaltzopoulou E, Villava CE, Bosch-Padrós M, Takata N, Trepat X, Eiraku M, Ebisuya M, (2022). Optogenetic control of apical constriction induces synthetic morphogenesis in mammalian tissues Nature Communications 13, 5400

The emerging field of synthetic developmental biology proposes bottom-up approaches to examine the contribution of each cellular process to complex morphogenesis. However, the shortage of tools to manipulate three-dimensional (3D) shapes of mammalian tissues hinders the progress of the field. Here we report the development of OptoShroom3, an optogenetic tool that achieves fast spatiotemporal control of apical constriction in mammalian epithelia. Activation of OptoShroom3 through illumination in an epithelial Madin-Darby Canine Kidney (MDCK) cell sheet reduces the apical surface of the stimulated cells and causes displacements in the adjacent regions. Light-induced apical constriction provokes the folding of epithelial cell colonies on soft gels. Its application to murine and human neural organoids leads to thickening of neuroepithelia, apical lumen reduction in optic vesicles, and flattening in neuroectodermal tissues. These results show that spatiotemporal control of apical constriction can trigger several types of 3D deformation depending on the initial tissue context.© 2022. The Author(s).

JTD Keywords: build, developmental biology, disease, light, localization, multicellular structures, organization, plate, shroom, Epithelial-cell shape


Zamora RA, López-Ortiz M, Sales-Mateo M, Hu C, Croce R, Maniyara RA, Pruneri V, Giannotti MI, Gorostiza P, (2022). Light- and Redox-Dependent Force Spectroscopy Reveals that the Interaction between Plastocyanin and Plant Photosystem I Is Favored when One Partner Is Ready for Electron Transfer Acs Nano 16, 15155-15164

Photosynthesis is a fundamental process that converts photons into chemical energy, driven by large protein complexes at the thylakoid membranes of plants, cyanobacteria, and algae. In plants, water-soluble plastocyanin (Pc) is responsible for shuttling electrons between cytochrome b6f complex and the photosystem I (PSI) complex in the photosynthetic electron transport chain (PETC). For an efficient turnover, a transient complex must form between PSI and Pc in the PETC, which implies a balance between specificity and binding strength. Here, we studied the binding frequency and the unbinding force between suitably oriented plant PSI and Pc under redox control using single molecule force spectroscopy (SMFS). The binding frequency (observation of binding-unbinding events) between PSI and Pc depends on their respective redox states. The interaction between PSI and Pc is independent of the redox state of PSI when Pc is reduced, and it is disfavored in the dark (reduced P700) when Pc is oxidized. The frequency of interaction between PSI and Pc is higher when at least one of the partners is in a redox state ready for electron transfer (ET), and the post-ET situation (PSIRed-PcOx) leads to lower binding. In addition, we show that the binding of ET-ready PcRed to PSI can be regulated externally by Mg2+ ions in solution.

JTD Keywords: architecture, binding-site, complexes, ferredoxin, force spectroscopy, induced structural-changes, interprotein electron transfer, light-dependent interaction, mg2+ concentration, photosystem i, plastocyanin, probe, recognition, reduction, Force spectroscopy, Interprotein electron transfer, Light-dependent interaction, Photosynthetic reaction-center, Photosystem i, Plastocyanin, Single molecule measurements


Casanellas I, Samitier J, Lagunas A, (2022). Recent advances in engineering nanotopographic substrates for cell studies Frontiers In Bioengineering And Biotechnology 10, 1002967

Cells sense their environment through the cell membrane receptors. Interaction with extracellular ligands induces receptor clustering at the nanoscale, assembly of the signaling complexes in the cytosol and activation of downstream signaling pathways, regulating cell response. Nanoclusters of receptors can be further organized hierarchically in the cell membrane at the meso- and micro-levels to exert different biological functions. To study and guide cell response, cell culture substrates have been engineered with features that can interact with the cells at different scales, eliciting controlled cell responses. In particular, nanoscale features of 1-100 nm in size allow direct interaction between the material and single cell receptors and their nanoclusters. Since the first "contact guidance" experiments on parallel microstructures, many other studies followed with increasing feature resolution and biological complexity. Here we present an overview of the advances in the field summarizing the biological scenario, substrate fabrication techniques and applications, highlighting the most recent developments.Copyright © 2022 Casanellas, Samitier and Lagunas.

JTD Keywords: cell response, density, differentiation, lithography, micro, nanofabrication, nanopatterning, nanopatterns, nanoscale, nanotopography, organization, photolithography, Cell response, Nanofabrication, Nanopatterning, Nanotopography, Plasma-membrane, Receptor nanoclustering


Fernández‐Costa, Juan M., Ortega, María A., Rodríguez‐Comas, Júlia, Lopez‐Muñoz, Gerardo, Yeste, Jose, Mangas‐Florencio, Lluís, Fernández‐González, Miriam, Martin‐Lasierra, Eduard, Tejedera‐Villafranca, Ainoa, Ramon‐Azcon, Javier, (2022). Training-on-a-Chip: A MultiOrgan Device to Study the Effect of Muscle Exercise on Insulin Secretion in Vitro Advanced Materials Technologies , 2200873

Riedelová, Zuzana, de los Santos Pereira, Andres, Svoboda, Jan, Pop‐Georgievski, Ognen, Májek, Pavel, Pečánková, Klára, Dyčka, Filip, Rodriguez‐Emmenegger, Cesar, Riedel, Tomáš, (2022). The Relation Between Protein Adsorption and Hemocompatibility of Antifouling Polymer Brushes Macromolecular Bioscience , 2200247

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


F Amil A, Rubio Ballester B, Maier M, FMJ Verschure P, (2022). Chronic use of cannabis might impair sensory error processing in the cerebellum through endocannabinoid dysregulation Addictive Behaviors 131, 107297

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, 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


Rizzello, L, De Matteis, V, (2022). Identification of SARS-CoV-2 by Gold Nanoparticles Biocell 46, 2369-2380

The SARS-CoV-2 outbreaks highlighted the need for effective, reliable, fast, easy-to-do and cheap diagnostics procedures. We pragmatically experienced that an early positive-case detection, inevitably coupled with a mass vaccination campaign, is a milestone to control the COVID-19 pandemic. Gold nanoparticles (AuNPs) can indeed play a crucial role in this context, as their physicochemical, optics and electronics properties are being extensively used in photothermal therapy (PTT), radiation therapy (RT), drug delivery and diagnostic. AuNPs can be synthesized by several approaches to obtain different sizes and shapes that can be easily functionalized with many kinds of molecules such as antibodies, proteins, probes, and lipids. In addition, AuNPs showed high biocompatibility making them useful tool in medicine field. We thus reviewed here the most relevant evidence on AuNPs as effective way to detect the presence of SARS-CoV-2 antigens. We trust future diagnostic efforts must take this 'old-fashioned' nanotechnology tool into consideration for the development and commercialization of reliable and feasible detection kits.

JTD Keywords: Aggregation, Antibodies, Assay, Covid-19, Diagnosis, Enhanced raman-scattering, Gold nanoparticles, Immunoassay, Pandemic disease, Physicochemical properties, Rapid detection, Sars-cov-2, Sensors, Surface-plasmon resonance, Therapy


Biosca, A, Ramirez, M, Gomez-Gomez, A, Lafuente, A, Iglesias, V, Pozo, OJ, Imperial, S, Fernandez-Busquets, X, (2022). Characterization of Domiphen Bromide as a New Fast-Acting Antiplasmodial Agent Inhibiting the Apicoplastidic Methyl Erythritol Phosphate Pathway Pharmaceutics 14, 1320

The evolution of resistance by the malaria parasite to artemisinin, the key component of the combination therapy strategies that are at the core of current antimalarial treatments, calls for the urgent identification of new fast-acting antimalarials. The apicoplast organelle is a preferred target of antimalarial drugs because it contains biochemical processes absent from the human host. Fosmidomycin is the only drug in clinical trials targeting the apicoplast, where it inhibits the methyl erythritol phosphate (MEP) pathway. Here, we characterized the antiplasmodial activity of domiphen bromide (DB), another MEP pathway inhibitor with a rapid mode of action that arrests the in vitro growth of Plasmodium falciparum at the early trophozoite stage. Metabolomic analysis of the MEP pathway and Krebs cycle intermediates in 20 mu M DB-treated parasites suggested a rapid activation of glycolysis with a concomitant decrease in mitochondrial activity, consistent with a rapid killing of the pathogen. These results present DB as a model compound for the development of new, potentially interesting drugs for future antimalarial combination therapies.

JTD Keywords: Antibiotics, Antimalarial drugs, Antimalarial-drug, Artemisinin, Combination therapies, Domiphen bromide, Intraerythrocytic stages, Isoprenoid biosynthesis, Malaria, Methyl erythritol phosphate pathway, Nonmevalonate pathway, Plasmodium falciparum, Plasmodium-falciparum apicoplast, Red-blood-cells, Targeted delivery


Lopez-Muñoz GA, Mughal S, Ramón-Azcón J, (2022). Sensors and Biosensors in Organs-on-a-Chip Platforms Microfluidics And Biosensors In Cancer Research 1379, 55-80

Biosensors represent a powerful analytical tool for analyzing biomolecular interactions with the potential to achieve real-time quantitative analysis with high accuracy using low sample volumes, minimum sample pretreatment with high potential for the development of in situ and highly integrated monitoring platforms. Considering these advantages, their use in cell-culture systems has increased over the last few years. Between the different technologies for cell culture, organs-on-a-chip (OOCs) represent a novel technology that tries to mimic an organ's functionality by combining tissue engineering/organoid with microfluidics. Although there are still challenges to achieving OOC models with high organ mimicking relevance, these devices can offer effective models for drug treatment development by identifying drug targets, screening toxicity, and determining the potential effects of drugs in living beings. Consequently, in the future, we might replace animal studies by offering more ethical test models. Considering the relevance that different physiological and biochemical parameters have in the correct functionality of cells, sensing and biosensing platforms can offer an effective way for the real-time monitoring of physiological parameters and, in our opinion, more relevant, the secretion of biomarkers such as cytokines, growth factors, and others related with the influence of drugs or other types of stimulus in cell metabolism. Keeping this concept in mind, in this chapter, we focus on describing the potential use of sensors and biosensors in OOC devices to achieve fully integrated platforms that monitor physiological parameters and cell metabolism.© 2022. The Author(s), under exclusive license to Springer Nature Switzerland AG.

JTD Keywords: Biosensors, Organ-on-a-chip, Pre-clinical platforms, Screening, Sensors


Yang, Bingquan, Wang, Yangxin, Vorobii, Mariia, Sauter, Eric, Koenig, Meike, Kumar, Ravi, Rodriguez-Emmenegger, Cesar, Hirtz, Michael, (2022). Evaluation of Dibenzocyclooctyne and Bicyclononyne Click Reaction on Azido-Functionalized Antifouling Polymer Brushes via Microspotting Advanced Materials Interfaces 9, 2102325

Bohner, Marc, Maazouz, Yassine, Ginebra, Maria-Pau, Habibovic, Pamela, Schoenecker, Jonathan G., Seeherman, Howard, van den Beucken, Jeroen, Witte, Frank, (2022). Sustained local ionic homeostatic imbalance caused by calcification modulates inflammation to trigger heterotopic ossification Acta Biomaterialia 145, 1-24

Heterotopic ossification (HO) is a condition triggered by an injury leading to the formation of mature lamellar bone in extraskeletal soft tissues. Despite being a frequent complication of orthopedic and trauma surgery, brain and spinal injury, the etiology of HO is poorly understood. The aim of this study is to evaluate the hypothesis that a sustained local ionic homeostatic imbalance (SLIHI) created by mineral formation during tissue calcification modulates inflammation to trigger HO. This evaluation also considers the role SLIHI could play for the design of cell-free, drug-free osteoinductive bone graft substitutes. The evaluation contains five main sections. The first section defines relevant concepts in the context of HO and provides a summary of proposed causes of HO. The second section starts with a detailed analysis of the occurrence and involvement of calcification in HO. It is followed by an explanation of the causes of calcification and its consequences. This allows to speculate on the potential chemical modulators of inflammation and triggers of HO. The end of this second section is devoted to in vitro mineralization tests used to predict the ectopic potential of materials. The third section reviews the biological cascade of events occurring during pathological and material-induced HO, and attempts to propose a quantitative timeline of HO formation. The fourth section looks at potential ways to control HO formation, either acting on SLIHI or on inflammation. Chemical, physical, and drug-based approaches are considered. Finally, the evaluation finishes with a critical assessment of the definition of osteoinduction.

JTD Keywords: beta-tricalcium phosphate, bone, bone graft, bone morphogenetic protein, demineralized bone-matrix, experimental myositis-ossificans, extracellular calcium, heterotopic ossification, in-vitro, inflammation, multinucleated giant-cells, osteoinduction, spinal-cord-injury, total hip-arthroplasty, traumatic brain-injury, Apatite, Calcium-sensing receptor


Raymond, Y, Lehmann, C, Thorel, E, Benitez, R, Riveiro, A, Pou, J, Manzanares, MC, Franch, J, Canal, C, Ginebra, MP, (2022). 3D printing with star-shaped strands: A new approach to enhance in vivo bone regeneration Biomaterials Advances 137, 212807

Concave surfaces have shown to promote bone regeneration in vivo. However, bone scaffolds obtained by direct ink writing, one of the most promising approaches for the fabrication of personalized bone grafts, consist mostly of convex surfaces, since they are obtained by microextrusion of cylindrical strands. By modifying the geometry of the nozzle, it is possible to print 3D structures composed of non-cylindrical strands and favor the presence of concave surfaces. In this work, we compare the in vivo performance of 3D-printed calcium phosphate scaffolds with either conventional cylindrical strands or star-shaped strands, in a rabbit femoral condyle model. Mono cortical defects, drilled in contralateral positions, are randomly grafted with the two scaffold configurations, with identical composition. The samples are explanted eight weeks post-surgery and assessed by ??-CT and resin embedded histological observations. The results reveal that the scaffolds containing star-shaped strands have better osteoconductive properties, guiding the newly formed bone faster towards the core of the scaffolds, and enhance bone regeneration, although the increase is not statistically significant (p > 0.05). This new approach represents a turning point towards the optimization of pore shape in 3D-printed bone grafts, further boosting the possibilities that direct ink writing technology offers for patient-specific applications.

JTD Keywords: 3d printing, Architecture, Biomimetic calcium phosphate, Bone regeneration, Calcium-phosphate scaffolds, Geometry, Growth, Implants, In vivo, Induction, Microporosity, Osteoinduction, Pore architecture, Scaffold, Surfaces, Tissue


Marhuenda, Esther, Villarino, Alvaro, Narciso, Maria Leonor, Camprubí-Rimblas, Marta, Farré, Ramon, Gavara, Núria, Artigas, Antonio, Almendros, Isaac, Otero, Jorge, (2022). Lung Extracellular Matrix Hydrogels Enhance Preservation of Type II Phenotype in Primary Alveolar Epithelial Cells International Journal Of Molecular Sciences 23, 4888

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, hydrogels, pathway, surfactant, type ii phenotype, yap, Extracellular matrix, Transplantation


Bonilla-Pons SÀ, Nakagawa S, Bahima EG, Fernández-Blanco Á, Pesaresi M, D'Antin JC, Sebastian-Perez R, Greco D, Domínguez-Sala E, Gómez-Riera R, Compte RIB, Dierssen M, Montserrat Pulido, N, Cosma MP, (2022). Müller glia fused with adult stem cells undergo neural differentiation in human retinal models Ebiomedicine 77, 103914

Visual impairments are a critical medical hurdle to be addressed in modern society. Müller glia (MG) have regenerative potential in the retina in lower vertebrates, but not in mammals. However, in mice, in vivo cell fusion between MG and adult stem cells forms hybrids that can partially regenerate ablated neurons.We used organotypic cultures of human retina and preparations of dissociated cells to test the hypothesis that cell fusion between human MG and adult stem cells can induce neuronal regeneration in human systems. Moreover, we established a microinjection system for transplanting human retinal organoids to demonstrate hybrid differentiation.We first found that cell fusion occurs between MG and adult stem cells, in organotypic cultures of human retina as well as in cell cultures. Next, we showed that the resulting hybrids can differentiate and acquire a proto-neural electrophysiology profile when the Wnt/beta-catenin pathway is activated in the adult stem cells prior fusion. Finally, we demonstrated the engraftment and differentiation of these hybrids into human retinal organoids.We show fusion between human MG and adult stem cells, and demonstrate that the resulting hybrid cells can differentiate towards neural fate in human model systems. Our results suggest that cell fusion-mediated therapy is a potential regenerative approach for treating human retinal dystrophies.This work was supported by La Caixa Health (HR17-00231), Velux Stiftung (976a) and the Ministerio de Ciencia e Innovación, (BFU2017-86760-P) (AEI/FEDER, UE), AGAUR (2017 SGR 689, 2017 SGR 926).Published by Elsevier B.V.

JTD Keywords: cell fusion, expression, fusion, ganglion-cells, in-vitro, mouse, müller glia, neural differentiation, organoids, regeneration, retina regeneration, stem cells, stromal cells, transplantation, 4',6 diamidino 2 phenylindole, 5' nucleotidase, Agarose, Alcohol, Arpe-19 cell line, Article, Beta catenin, Beta tubulin, Bone-marrow-cells, Bromophenol blue, Buffer, Calcium cell level, Calcium phosphate, Calretinin, Canonical wnt signaling, Cd34 antigen, Cell culture, Cell fusion, Cell viability, Coculture, Complementary dna, Confocal microscopy, Cornea transplantation, Cryopreservation, Cryoprotection, Crystal structure, Current clamp technique, Dimethyl sulfoxide, Dodecyl sulfate sodium, Edetic acid, Electrophysiology, Endoglin, Fetal bovine serum, Fibroblast growth factor 2, Flow cytometry, Fluorescence activated cell sorting, Fluorescence intensity, Glyceraldehyde 3 phosphate dehydrogenase, Glycerol, Glycine, Hoe 33342, Immunofluorescence, Immunohistochemistry, Incubation time, Interleukin 1beta, Lentivirus vector, Matrigel, Mercaptoethanol, Microinjection, Mueller cell, Müller glia, N methyl dextro aspartic acid, Nerve cell differentiation, Neural differentiation, Nitrogen, Nonhuman, Organoids, Paraffin, Paraffin embedding, Paraformaldehyde, Patch clamp technique, Penicillin derivative, Phenolsulfonphthalein, Phenotype, Phosphate buffered saline, Phosphoprotein phosphatase inhibitor, Polyacrylamide gel electrophoresis, Potassium chloride, Povidone iodine, Promoter region, Proteinase inhibitor, Real time polymerase chain reaction, Receptor type tyrosine protein phosphatase c, Restriction endonuclease, Retina, Retina dystrophy, Retina regeneration, Retinol, Rhodopsin, Rna extraction, Stem cell, Stem cells, Subcutaneous fat, Tunel assay, Visual impairment, Western blotting


Hüttener, Mário, Hergueta, Jon, Bernabeu, Manuel, Prieto, Alejandro, Aznar, Sonia, Merino, Susana, Tomás, Joan, Juárez, Antonio, (2022). Roles of Proteins Containing Immunoglobulin-Like Domains in the Conjugation of Bacterial Plasmids Msphere 7, e00978-21

Transmission of a plasmid from one bacterial cell to another, in several instances, underlies the dissemination of antimicrobial resistance (AMR) genes. The process requires well-characterized enzymatic machinery that facilitates cell-to-cell contact and the transfer of the plasmid.

JTD Keywords: antimicrobial resistance, bacterial ig-like proteins, bacterial lg-like proteins, chromosomal genes, identification, inca/c, mutational analysis, plasmid conjugation, products, r-factors, resistance plasmids, salmonella-enterica, sequence, Antimicrobial resistance, Bacterial ig-like proteins, Escherichia-coli, Plasmid conjugation


Georgiev VN, Avalos-Padilla Y, Fernàndez-Busquets X, Dimova R, (2022). Femtoliter Injection of ESCRT-III Proteins into Adhered Giant Unilamellar Vesicles Bio Protoc 12, e4328

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, light, microinjection, microparticles, plasma, Adhesion, Budding, Escrt-iii, Extracellular vesicles, Giant unilamellar vesicle (guv), Membrane, Microinjection


Raymond, Yago, Johansson, Linh, Thorel, Emilie, Ginebra, Maria-Pau, (2022). Translation of three-dimensional printing of ceramics in bone tissue engineering and drug delivery Mrs Bulletin 47, 59-69

López Ortiz, Manuel, Zamora, Ricardo A., Giannotti, Marina Inés, Hu, Chen, Croce, Roberta, Gorostiza, Pau, (2022). Distance and Potential Dependence of Charge Transport Through the Reaction Center of Individual Photosynthetic Complexes Small 18, 2104366

Charge separation and transport through the reaction center of photosystem I (PSI) is an essential part of the photosynthetic electron transport chain. A strategy is developed to immobilize and orient PSI complexes on gold electrodes allowing to probe the complex's electron acceptor side, the chlorophyll special pair P700. Electrochemical scanning tunneling microscopy (ECSTM) imaging and current-distance spectroscopy of single protein complex shows lateral size in agreement with its known dimensions, and a PSI apparent height that depends on the probe potential revealing a gating effect in protein conductance. In current-distance spectroscopy, it is observed that the distance-decay constant of the current between PSI and the ECSTM probe depends on the sample and probe electrode potentials. The longest charge exchange distance (lowest distance-decay constant ?) is observed at sample potential 0 mV/SSC (SSC: reference electrode silver/silver chloride) and probe potential 400 mV/SSC. These potentials correspond to hole injection into an electronic state that is available in the absence of illumination. It is proposed that a pair of tryptophan residues located at the interface between P700 and the solution and known to support the hydrophobic recognition of the PSI redox partner plastocyanin, may have an additional role as hole exchange mediator in charge transport through PSI.© 2021 Wiley-VCH GmbH.

JTD Keywords: azurin, current distance decay spectroscopy, cytochrome c(6), electrochemical scanning tunneling microscopy (ecstm), electrochemistry, photosystem i, photosystem-i, plastocyanin, protein electron transfer, recognition, single metalloprotein, single molecules, structural basis, tunneling spectroscopy, 'current, Amino acids, Charge transfer, Chlorine compounds, Current distance decay spectroscopy, Decay spectroscopies, Distance decay, Electrochemical scanning tunneling microscopy, Electrochemical scanning tunneling microscopy (ecstm), Electrodes, Electron transfer, Electron transport properties, Gold compounds, Photosystem i, Photosystems, Protein electron transfer, Protein electron-transfer, Proteins, Scanning tunneling microscopy, Silver halides, Single molecule, Single molecules


Bravo, J, Ribeiro, I, Terceiro, AF, Andrade, EB, Portugal, CC, Lopes, IM, Azevedo, MM, Sousa, M, Lopes, CDF, Lobo, AC, Canedo, T, Relvas, JB, Summavielle, T, (2022). Neuron-Microglia Contact-Dependent Mechanisms Attenuate Methamphetamine-Induced Microglia Reactivity and Enhance Neuronal Plasticity Cells 11, 355

Exposure to methamphetamine (Meth) has been classically associated with damage to neuronal terminals. However, it is now becoming clear that addiction may also result from the interplay between glial cells and neurons. Recently, we demonstrated that binge Meth administration promotes microgliosis and microglia pro-inflammation via astrocytic glutamate release in a TNF/IP(3)R2-Ca2+-dependent manner. Here, we investigated the contribution of neuronal cells to this process. As the crosstalk between microglia and neurons may occur by contact-dependent and/or contact-independent mechanisms, we developed co-cultures of primary neurons and microglia in microfluidic devices to investigate how their interaction affects Meth-induced microglia activation. Our results show that neurons exposed to Meth do not activate microglia in a cell-autonomous way but require astrocyte mediation. Importantly, we found that neurons can partially prevent Meth-induced microglia activation via astrocytes, which seems to be achieved by increasing arginase 1 expression and strengthening the CD200/CD200r pathway. We also observed an increase in synaptic individual area, as determined by co-localization of pre- and post-synaptic markers. The present study provides evidence that contact-dependent mechanisms between neurons and microglia can attenuate pro-inflammatory events such as Meth-induced microglia activation.

JTD Keywords: cd200, contact-dependent, methamphetamine, neuron-to-microglia, psd95, Activation, Cd200, Contact-dependent, Expression, Glutamate, Methamphetamine, Neuron-to-microglia, Neuroprotection, Platform, Psd95


Boda, SK, Aparicio, C, (2022). Dual keratinocyte-attachment and anti-inflammatory coatings for soft tissue sealing around transmucosal oral implants Biomaterials Science 10, 665-677

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


Páscoa dos Santos F, Verschure PFMJ, (2022). Excitatory-Inhibitory Homeostasis and Diaschisis: Tying the Local and Global Scales in the Post-stroke Cortex Frontiers In Systems Neuroscience 15, 806544

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: 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, Visual cortex


Dulay, S, Rivas, L, Pla, L, Berdun, S, Eixarch, E, Gratacos, E, Illa, M, Mir, M, Samitier, J, (2021). Fetal ischemia monitoring with in vivo implanted electrochemical multiparametric microsensors Journal Of Biological Engineering 15, 28

Under intrauterine growth restriction (IUGR), abnormal attainment of the nutrients and oxygen by the fetus restricts the normal evolution of the prenatal causing in many cases high morbidity being one of the top-ten causes of neonatal death. The current gold standards in hospitals to detect this relevant problem is the clinical observation by echography, cardiotocography and Doppler. These qualitative techniques are not conclusive and requires risky invasive fetal scalp blood testing and/or amniocentesis. We developed micro-implantable multiparametric electrochemical sensors for measuring ischemia in real time in fetal tissue and vascular. This implantable technology is designed to continuous monitoring for an early detection of ischemia to avoid potential fetal injury. Two miniaturized electrochemical sensors were developed based on oxygen and pH detection. The sensors were optimized in vitro under controlled concentration, to assess the selectivity and sensitivity required. The sensors were then validated in vivo in the ewe fetus model, by means of their insertion in the muscle leg and inside the iliac artery of the fetus. Ischemia was achieved by gradually obstructing the umbilical cord to regulate the amount of blood reaching the fetus. An important challenge in fetal monitoring is the detection of low levels of oxygen and pH changes under ischemic conditions, requiring high sensitivity sensors. Significant differences were observed in both; pH and pO(2) sensors under changes from normoxia to hypoxia states in the fetus tissue and vascular with both sensors. Herein, we demonstrate the feasibility of the developed sensors for future fetal monitoring in medical applications.

JTD Keywords: electrochemical biosensor, implantable sensor, in vivo validation, ischemia detection, tissue and vascular monitoring, Animal experiment, Animal model, Animal tissue, Article, Blood-gases, Brain, Classification, Controlled study, Diagnosis, Doppler, Early diagnosis, Electrochemical analysis, Electrochemical biosensor, Ewe, Feasibility study, Female, Fetus, Fetus disease, Fetus monitoring, Gestational age, Hypoxemia, Iliac artery, Implantable sensor, In vivo validation, Intrauterine growth restriction, Intrauterine growth retardation, Ischemia detection, Leg muscle, Management, Nonhuman, Oxygen consumption, Ph, Ph and oxygen detection, Ph measurement, Process optimization, Sheep, Tissue and vascular monitoring, Umbilical-cord occlusion


Duro-Castano, Aroa, Rodríguez-Arco, Laura, Ruiz-Pérez, Lorena, De Pace, Cesare, Marchello, Gabriele, Noble-Jesus, Carlos, Battaglia, Giuseppe, (2021). One-Pot Synthesis of Oxidation-Sensitive Supramolecular Gels and Vesicles Biomacromolecules 22, 5052-5064

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, One-pot synthesis, Organic solvents, Oxidation, Physiological condition, Polyethylene oxides, Post-polymerization, Ring-opening polymerization, Scaffolds (biology), Self assembly, Stimuli-responsive properties, Supramolecular chemistry, Supramolecular gels, Supramolecular micelles, Wormlike micelle


Vila, JC, Castro-Aguirre, N, Lopez-Munoz, GA, Ferret-Minana, A, De Chiara, F, Ramon-Azcon, J, (2021). Disposable Polymeric Nanostructured Plasmonic Biosensors for Cell Culture Adhesion Monitoring Frontiers In Bioengineering And Biotechnology 9, 799325

Over the last years, optical biosensors based on plasmonic nanomaterials have gained great scientific interest due to their unquestionable advantages compared to other biosensing technologies. They can achieve sensitive, direct, and label-free analysis with exceptional potential for multiplexing and miniaturization. Recently, it has been demonstrated the potential of using optical discs as high throughput nanotemplates for the development of plasmonic biosensors in a cost-effective way. This work is a pilot study focused on the development of an integrated plasmonic biosensor for the monitoring of cell adhesion and growth of human retinal pigmented cell line (ARPE-19) under different media conditions (0 and 2% of FBS). We observed an increase of the plasmonic band displacement under 2% FBS compared to 0% conditions over time (1, 3, and 5 h). These preliminary results show that the proposed plasmonic biosensing approach is a direct, non-destructive, and real-time tool that could be employed in the study of living cells behavior and culture conditions. Furthermore, this setup could assess the viability of the cells and their growth over time with low variability between the technical replicates improving the experimental replicability.

JTD Keywords: cell confluency, cell culture, nanocrystals, optical biosensor, Adhesion monitoring, Biosensing, Biosensors, Cell adhesion, Cell confluency, Cell culture, Cells, Condition, Cost effectiveness, Disposables, Nano-structured, Nanocrystals, Optical bio-sensors, Optical biosensor, Plasmonic biosensors, Plasmonic nanostructures, Plasmonics, Polylysine


Chacon DS, Torres TM, da Silva IB, de Araújo TF, Roque AdA, Pinheiro FASD, Selegato D, Pilon A, Reginaldo FPS, da Costa CT, Vilasboa J, Freire RT, Voigt EL, Zuanazzi JAS, Libonati R, Rodrigues JA, Santos FLM, Scortecci KC, Lopes NP, Ferreira LDS, dos Santos LV, Cavalheiro AJ, Fett-Neto AG, Giordani RB, (2021). Erythrina velutina Willd. alkaloids: Piecing biosynthesis together from transcriptome analysis and metabolite profiling of seeds and leaves Journal Of Advanced Research 34, 123-136

© 2021 Introduction: Natural products of pharmaceutical interest often do not reach the drug market due to the associated low yields and difficult extraction. Knowledge of biosynthetic pathways is a key element in the development of biotechnological strategies for plant specialized metabolite production. The scarce studies regarding non-model plants impair advances in this field. Erythrina spp. are mainly used as central nervous system depressants in folk medicine and are important sources of bioactive tetracyclic benzylisoquinoline alkaloids, which can act on several pathology-related biological targets. Objective: Herein the purpose is to employ combined transcriptome and metabolome analyses (seeds and leaves) of a non-model medicinal Fabaceae species grown in its unique arid natural habitat. The study tries to propose a putative biosynthetic pathway for the bioactive alkaloids by using an omic integrated approach. Methods: The Next Generation Sequencing-based transcriptome (de novo RNA sequencing) was carried out in a Illumina NextSeq 500 platform. Regarding the targeted metabolite profiling, Nuclear Magnetic Resonance and the High-Performance Liquid Chromatography coupled to a micrOTOF-QII, High Resolution Mass Spectrometer, were used. Results: This detailed macro and micromolecular approach applied to seeds and leaves of E. velutina revealed 42 alkaloids by metabolome tools. Based on the combined evidence, 24 gene candidates were put together in a putative pathway leading to the singular alkaloid diversity of this species. Conclusion: These results contribute by indicating potential biotechnological targets Erythrina alkaloids biosynthesis as well as to improve molecular databases with omic data from a non-model medicinal plant. Furthermore, they reveal an interesting chemical diversity in Erythrina velutina harvested in Caatinga. Last, but not least, this data may also contribute to tap Brazilian biodiversity in a rational and sustainable fashion, promoting adequate public policies for preservation and protection of sensitive areas within the Caatinga.

JTD Keywords: benzylisoquinoline alkaloids, caatinga, codeinone reductase, erythrina velutina, expression, mass-spectrometry, molecular-cloning, morphine biosynthesis, natural-products, opium poppy, papaver-somniferum, plant-metabolism, targeted metabolite profile, transcriptome, Benzylisoquinoline alkaloids, Berberine bridge enzyme, Caatinga, Erythrina velutina, Targeted metabolite profile, Transcriptome


García-Mintegui C, Córdoba LC, Buxadera-Palomero J, Marquina A, Jiménez-Piqué E, Ginebra MP, Cortina JL, Pegueroles M, (2021). Zn-Mg and Zn-Cu alloys for stenting applications: From nanoscale mechanical characterization to in vitro degradation and biocompatibility Bioactive Materials 6, 4430-4446

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


Lopez-Muñoz, Gerardo A, Fernández-Costa, Juan M, Ortega, Maria Alejandra, Balaguer-Trias, Jordina, Martin-Lasierra, Eduard, Ramón-Azcón, Javier, (2021). Plasmonic nanocrystals on polycarbonate substrates for direct and label-free biodetection of Interleukin-6 in bioengineered 3D skeletal muscles Nanophotonics 10, 4477-4488

Abstract The development of nanostructured plasmonic biosensors has been widely widespread in the last years, motivated by the potential benefits they can offer in integration, miniaturization, multiplexing opportunities, and enhanced performance label-free biodetection in a wide field of applications. Between them, engineering tissues represent a novel, challenging, and prolific application field for nanostructured plasmonic biosensors considering the previously described benefits and the low levels of secreted biomarkers (?pM–nM) to detect. Here, we present an integrated plasmonic nanocrystals-based biosensor using high throughput nanostructured polycarbonate substrates. Metallic film thickness and incident angle of light for reflectance measurements were optimized to enhance the detection of antibody–antigen biorecognition events using numerical simulations. We achieved an enhancement in biodetection up to 3× as the incident angle of light decreases, which can be related to shorter evanescent decay lengths. We achieved a high reproducibility between channels with a coefficient of variation below 2% in bulk refractive index measurements, demonstrating a high potential for multiplexed sensing. Finally, biosensing potential was demonstrated by the direct and label-free detection of interleukin-6 biomarker in undiluted cell culture media supernatants from bioengineered 3D skeletal muscle tissues stimulated with different concentrations of endotoxins achieving a limit of detection (LOD) of ? 0.03 ng/mL (1.4 pM).

JTD Keywords: assay, crystals, drug, label-free biosensing, molecules, plasmonic nanostructures, sensors, skeletal muscle, tissue engineering, Biodetection, Biomarkers, Biosensors, Cell culture, Cells, Chemical detection, Histology, Interleukin-6, Interleukin6 (il6), Label free, Label-free biosensing, Muscle, Nano-structured, Nanocrystals, Plasmonic nanocrystals, Plasmonic nanostructures, Plasmonics, Polycarbonate substrates, Polycarbonates, Refractive index, Sensitivity, Skeletal muscle, Tissue engineering, Tissues engineerings


Garreta, E, Nauryzgaliyeva, Z, Montserrat, N, (2021). Human induced pluripotent stem cell-derived kidney organoids toward clinical implementations Curr Opin Biomed Eng 20, 100346

The generation of kidney organoids from human pluripotent stem cells (hPSCs) has represented a relevant scientific achievement in the organoid field. Importantly, hPSC-derived kidney organoids contain multiple nephron-like structures that exhibit some renal functional characteristics and have the capacity to respond to nephrotoxic agents. In this review, we first discuss how bioengineering approaches can help overcome current kidney organoid challenges. Next, we focus on recent works exploiting kidney organoids for drug screening and disease modeling applications. Finally, we provide a state of the art on current research toward the potential application of kidney organoids and renal cells derived from hPSCs for future renal replacement therapies.

JTD Keywords: Bioengineering, Converting enzyme-ii, Crispr/cas9 gene editing, Disease, Disease modeling, Extracellular-matrix, Generation, Human pluripotent stem cells, Kidney organoids, Kidney regeneration, Model, Mouse, Reveals, Scaffold, Transplantation


Guasch-Girbau A, Fernàndez-Busquets X, (2021). Review of the current landscape of the potential of nanotechnology for future malaria diagnosis, treatment, and vaccination strategies Pharmaceutics 13, 2189

Malaria eradication has for decades been on the global health agenda, but the causative agents of the disease, several species of the protist parasite Plasmodium, have evolved mechanisms to evade vaccine-induced immunity and to rapidly acquire resistance against all drugs entering clinical use. Because classical antimalarial approaches have consistently failed, new strategies must be explored. One of these is nanomedicine, the application of manipulation and fabrication technology in the range of molecular dimensions between 1 and 100 nm, to the development of new medical solutions. Here we review the current state of the art in malaria diagnosis, prevention, and therapy and how nanotechnology is already having an incipient impact in improving them. In the second half of this review, the next generation of antimalarial drugs currently in the clinical pipeline is presented, with a definition of these drugs’ target product profiles and an assessment of the potential role of nanotechnology in their development. Opinions extracted from interviews with experts in the fields of nanomedicine, clinical malaria, and the economic landscape of the disease are included to offer a wider scope of the current requirements to win the fight against malaria and of how nanoscience can contribute to achieve them. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

JTD Keywords: antibody-bearing liposomes, antimalarial drugs, combination therapies, drug-delivery strategies, malaria diagnosis, malaria prophylaxis, malaria therapy, nanocarriers, nanomedicine, nanoparticles, nanotechnology, plasmodium, plasmodium-falciparum, red-blood-cells, targeted delivery, targeted drug delivery, vitro antimalarial activity, Antimalarial drugs, Isothermal amplification lamp, Malaria diagnosis, Malaria prophylaxis, Malaria therapy, Nanocarriers, Nanomedicine, Nanotechnology, Plasmodium, Targeted drug delivery


Nashimoto Y, Abe M, Fujii R, Taira N, Ida H, Takahashi Y, Ino K, Ramon-Azcon J, Shiku H, (2021). Topography and Permeability Analyses of Vasculature-on-a-Chip Using Scanning Probe Microscopies Advanced Healthcare Materials 10, 2101186

Microphysiological systems (MPS) or organs-on-chips (OoC) can emulate the physiological functions of organs in vitro and are effective tools for determining human drug responses in preclinical studies. However, the analysis of MPS has relied heavily on optical tools, resulting in difficulties in real-time and high spatial resolution imaging of the target cell functions. In this study, the role of scanning probe microscopy (SPM) as an analytical tool for MPS is evaluated. An access hole is made in a typical MPS system with stacked microchannels to insert SPM probes into the system. For the first study, a simple vascular model composed of only endothelial cells is prepared for SPM analysis. Changes in permeability and local chemical flux are quantitatively evaluated during the construction of the vascular system. The morphological changes in the endothelial cells after flow stimulation are imaged at the single-cell level for topographical analysis. Finally, the possibility of adapting the permeability and topographical analysis using SPM for the intestinal vascular system is further evaluated. It is believed that this study will pave the way for an in situ permeability assay and structural analysis of MPS using SPM.

JTD Keywords: cell, electrochemical microscopy, membrane-permeability, microphysiological systems, organs-chips, platform, scanning electrochemical microscopy, scanning ion conductance microscopy, scanning probe microscopy, shear-stress, surface-topography, Ion conductance microscopy, Microphysiological systems, Organs-chips, Scanning electrochemical microscopy, Scanning ion conductance microscopy, Scanning probe microscopy


Avalos-Padilla Y, Georgiev VN, Dimova R, (2021). ESCRT-III induces phase separation in model membranes prior to budding and causes invagination of the liquid-ordered phase Biochimica et Biophysica Acta - Biomembranes 1863, 183689

Membrane fission triggered by the endosomal sorting complex required for transport (ESCRT) is an important process observed in several pathogenic and non-pathogenic cellular events. From a synthetic-biology viewpoint, ESCRT proteins represent an interesting machinery for the construction of cell mimetic sub-compartments produced by fission. Since their discovery, the studies on ESCRT-III-mediated action, have mainly focused on protein dynamics, ignoring the role of lipid organization and membrane phase state. Recently, it has been suggested that membrane buds formed by the action of ESCRT-III are generated from transient microdomains in endosomal membranes. However, the interplay between membrane domain formation and ESCRT remodeling pathways has not been investigated. Here, giant unilamellar vesicles made of ternary lipid mixtures, either homogeneous in phase or exhibiting liquid-ordered/liquid-disordered phase coexistence, were employed as a model membrane system. These vesicles were incubated with purified recombinant ESCRT-III proteins from the parasite Entamoeba histolytica. In homogeneous membranes, we observe that EhVps32 can trigger domain formation while EhVps20 preferentially co-localizes in the liquid disordered phase. The addition of EhVps24 appears to induce the formation of intraluminal vesicles produced from the liquid-ordered phase. In phase separated membranes, the intraluminal vesicles are also generated from the liquid-ordered phase and presumably emerge from the phase boundary region. Our findings reinforce the hypothesis that ESCRT-mediated remodeling depends on the membrane phase state. Furthermore, the obtained results point to a potential synthetic biology approach for establishing eukaryotic mimics of artificial cells with microcompartments of specific membrane composition, which can also differ from that of the mother vesicle.

JTD Keywords: cell-membranes, coexistence, complex, escrt-iii, fission, guvs, lipid domains, lipid rafts, membrane fission, microcompartments, microscopy, phase separation, plasma-membrane, protein microarrays, structural basis, ternary mixtures, Escrt-iii, Giant unilamellar vesicles, Guvs, Lipid domains, Membrane fission, Microcompartments, Phase separation, Ternary mixtures


Mestre R, García N, Patiño T, Guix M, Fuentes J, Valerio-Santiago M, Almiñana N, Sánchez S, (2021). 3D-bioengineered model of human skeletal muscle tissue with phenotypic features of aging for drug testing purposes Biofabrication 13, 45011

Three-dimensional engineering of skeletal muscle is becoming increasingly relevant for tissue engineering, disease modeling and bio-hybrid robotics, where flexible, versatile and multidisciplinary approaches for the evaluation of tissue differentiation, functionality and force measurement are required. This works presents a 3D-printed platform of bioengineered human skeletal muscle which can efficiently model the three-dimensional structure of native tissue, while providing information about force generation and contraction profiles. Proper differentiation and maturation of myocytes is demonstrated by the expression of key myo-proteins using immunocytochemistry and analyzed by confocal microscopy, and the functionality assessed via electrical stimulation and analysis of contraction kinetics. To validate the flexibility of this platform for complex tissue modeling, the bioengineered muscle is treated with tumor necrosis factor α to mimic the conditions of aging, which is supported by morphological and functional changes. Moreover, as a proof of concept, the effects of Argireline® Amplified peptide, a cosmetic ingredient that causes muscle relaxation, are evaluated in both healthy and aged tissue models. Therefore, the results demonstrate that this 3D-bioengineered human muscle platform could be used to assess morphological and functional changes in the aging process of muscular tissue with potential applications in biomedicine, cosmetics and bio-hybrid robotics.

JTD Keywords: 3d bioprinting, bio-actuator, drug testing, human skeletal muscle, muscle ageing, platform, tnf-alpha, 3d bioprinting, Bio-actuator, Drug testing, Human skeletal muscle, Muscle ageing, Necrosis-factor-alpha


Rodríguez-Contreras A, Torres D, Rafik B, Ortiz-Hernandez M, Ginebra MP, Calero JA, Manero JM, Ruperez E, (2021). Bioactivity and antibacterial properties of calcium- and silver-doped coatings on 3D printed titanium scaffolds Surface & Coatings Technology 421

One of the major problems faced by metallic implants is the high probability of bacterial infections, with significant consequences for the patient. In this work, a thermochemical treatment is proposed to obtain silver-doped calcium titanate coatings on the Ti surface to improve the bioactivity of porous 3D-printed Ti structures and simultaneously provide them with antibacterial properties. A complete characterization of the new coating, the study of the ion release and the analysis of its cytotoxicity were carried out together with evaluation of the natural apatite forming in simulated body fluid (SBF). Moreover, the antibacterial properties of the coatings were assessed against Pseudomona aeruginosa and Escherichia coli as gram-negative and Staphylococcus aureus and Staphylococcus epidermidis as gram-positive bacterial strains. Ag ions were integrated into the Ca titanate layer and Ag nanoparticles were formed within the entire 3D Ti surface. Ca and Ag ions were released from both porous and solid samples into the Hanks' solution for 48 h. The treated surfaces showed no cytotoxicity and an apatite layer precipitated on the entire porous surface when the samples were immersed in SBF. The release of Ag from the surface had a strong antibacterial effect and prevented bacterial adhesion and proliferation on the surface. Moreover, the nanostructured topography of the coating resulted also in a reduction of bacterial adhesion and proliferation, even in absence of Ag. In conclusion, the cost-effective approach here reported provided protection against the most predominant bacterial colonizers to the Ti porous implants, while maintaining their bioactivity.

JTD Keywords: 3d-printing, alkaline, antibacterial activity, arthroplasty, bacterial adhesion, biomaterials, generation, ions, nanoparticles, osseointegration, silver, surface-layer, titanium implants, toxicity, 3d-printing, Antibacterial activity, Biomaterials, Porous structures, Silver, Ti metal, Titanium implants


Pilat N, Lefsihane K, Brouard S, Kotsch K, Falk C, Steiner R, Thaunat O, Fusil F, Montserrat N, Amarelli C, Casiraghi F, (2021). T- and B-cell therapy in solid organ transplantation: current evidence and future expectations Transplant International 34, 1594-1606

Cell therapy has emerged as an attractive therapeutic option in organ transplantation. During the last decade, the therapeutic potency of Treg immunotherapy has been shown in various preclinical animal models and safety was demonstrated in first clinical trials. However, there are still critical open questions regarding specificity, survival, and migration to the target tissue so the best Treg population for infusion into patients is still under debate. Recent advances in CAR technology hold the promise for Treg-functional superiority. Another exciting strategy is the generation of B-cell antibody receptor (BAR) Treg/cytotoxic T cells to specifically regulate or deplete alloreactive memory B cells. Finally, B cells are also capable of immune regulation, making them promising candidates for immunomodulatory therapeutic strategies. This article summarizes available literature on cell-based innovative therapeutic approaches aiming at modulating alloimmune response for transplantation. Crucial areas of investigation that need a joined effort of the transplant community for moving the field toward successful achievement of tolerance are highlighted.

JTD Keywords: allograft, autoimmune, b-cell antibody receptor t cells, chimeric antigen receptor tregs, expansion, expression, identification, infectious tolerance, mouse, prevention, regulatory b cells, regulatory t cells, signature, B-cell antibody receptor t cells, Chimeric antigen receptor tregs, Kidney-transplantation, Regulatory b cells, Regulatory t cells


Hamouda I, Labay C, Cvelbar U, Ginebra MP, Canal C, (2021). Selectivity of direct plasma treatment and plasma-conditioned media in bone cancer cell lines Scientific Reports 11, 17521

Atmospheric pressure plasma jets have been shown to impact several cancer cell lines, both in vitro and in vivo. These effects are based on the biochemistry of the reactive oxygen and nitrogen species generated by plasmas in physiological liquids, referred to as plasma-conditioned liquids. Plasma-conditioned media are efficient in the generation of reactive species, inducing selective cancer cell death. However, the concentration of reactive species generated by plasma in the cell culture media of different cell types can be highly variable, complicating the ability to draw precise conclusions due to the differential sensitivity of different cells to reactive species. Here, we compared the effects of direct and indirect plasma treatment on non-malignant bone cells (hOBs and hMSCs) and bone cancer cells (SaOs-2s and MG63s) by treating the cells directly or exposing them to previously treated cell culture medium. Biological effects were correlated with the concentrations of reactive species generated in the liquid. A linear increase in reactive species in the cell culture medium was observed with increased plasma treatment time independent of the volume treated. Values up to 700 µM for H2O2 and 140 µM of NO2− were attained in 2 mL after 15 min of plasma treatment in AdvDMEM cell culture media. Selectivity towards bone cancer cells was observed after both direct and indirect plasma treatments, leading to a decrease in bone cancer cell viability at 72 h to 30% for the longest plasma treatment times while maintaining the survival of non-malignant cells. Therefore, plasma-conditioned media may represent the basis for a potentially novel non-invasive technique for bone cancer therapy.

JTD Keywords: expression, in-vitro, jet, mechanisms, nitrate, nitrite, osteosarcoma cells, reactive oxygen, Cold atmospheric plasma


Alcaraz J, Ikemori R, Llorente A, Díaz-valdivia N, Reguart N, Vizoso M, (2021). Epigenetic reprogramming of tumor-associated fibroblasts in lung cancer: Therapeutic opportunities Cancers 13, 3782

Lung cancer is the leading cause of cancer-related death worldwide. The desmoplastic stroma of lung cancer and other solid tumors is rich in tumor-associated fibroblasts (TAFs) exhibiting an activated/myofibroblast-like phenotype. There is growing awareness that TAFs support key steps of tumor progression and are epigenetically reprogrammed compared to healthy fibroblasts. Although the mechanisms underlying such epigenetic reprogramming are incompletely understood, there is increasing evidence that they involve interactions with either cancer cells, pro-fibrotic cytokines such as TGF-β, the stiffening of the surrounding extracellular matrix, smoking cigarette particles and other environmental cues. These aberrant interactions elicit a global DNA hypomethylation and a selective transcriptional repression through hypermethylation of the TGF-β transcription factor SMAD3 in lung TAFs. Likewise, similar DNA methylation changes have been reported in TAFs from other cancer types, as well as histone core modifications and altered microRNA expression. In this review we summarize the evidence of the epigenetic reprogramming of TAFs, how this reprogramming contributes to the acquisition and maintenance of a tumor-promoting phenotype, and how it provides novel venues for therapeutic intervention, with a special focus on lung TAFs.

JTD Keywords: cancer-associated fibroblasts, desmoplasia, dna methylation, epigenetics, expression, genomic dna, lung cancer, mechanical memory, myofibroblast differentiation, pulmonary fibroblasts, smoking, stromal fibroblasts, tgf-?, tgf-beta, transforming growth-factor-beta-1, tumor stroma, Cancer-associated fibroblasts, Carcinoma-associated fibroblasts, Desmoplasia, Epigenetics, Lung cancer, Smoking, Tgf-β, Tumor stroma


Villasante A, Robinson ST, Cohen AR, Lock R, Guo XE, Vunjak-Novakovic G, (2021). Human Serum Enhances Biomimicry of Engineered Tissue Models of Bone and Cancer Frontiers In Bioengineering And Biotechnology 9, 658472

For decades, fetal bovine serum (FBS) has been used routinely for culturing many cell types, based on its empirically demonstrated effects on cell growth, and the lack of suitable non-xenogeneic alternatives. The FBS-based culture media do not represent the human physiological conditions, and can compromise biomimicry of preclinical models. To recapitulate in vitro the features of human bone and bone cancer, we investigated the effects of human serum and human platelet lysate on modeling osteogenesis, osteoclastogenesis, and bone cancer in two-dimensional (2D) and three-dimensional (3D) settings. For monitoring tumor growth within tissue-engineered bone in a non-destructive fashion, we generated cancer cell lines expressing and secreting luciferase. Culture media containing human serum enhanced osteogenesis and osteoclasts differentiation, and provided a more realistic in vitro mimic of human cancer cell proliferation. When human serum was used for building 3D engineered bone, the tissue recapitulated bone homeostasis and response to bisphosphonates observed in native bone. We found disparities in cell behavior and drug responses between the metastatic and primary cancer cells cultured in the bone niche, with the effectiveness of bisphosphonates observed only in metastatic models. Overall, these data support the utility of human serum for bioengineering of bone and bone cancers.

JTD Keywords: 3d cancer models, 3rs, alpha tnf-alpha, culture, cypridina luciferase, ewings-sarcoma, ewing’s sarcoma, human platelet lysate, human serum, human tumor, in-vitro, osteogenic differentiation, stem-cells, zoledronic acid, 3d cancer models, 3rs, Cypridina luciferase, Ewing's sarcoma, Ewing’s sarcoma, Fetal bovine serum, Human serum


Mendoza MB, Gutierrez S, Ortiz R, Moreno DF, Dermit M, Dodel M, Rebollo E, Bosch M, Mardakheh FK, Gallego C, (2021). The elongation factor eEF1A2 controls translation and actin dynamics in dendritic spines Science Signaling 14, eabf5594

Synaptic plasticity involves structural modifications in dendritic spines that are modulated by local protein synthesis and actin remodeling. Here, we investigated the molecular mechanisms that connect synaptic stimulation to these processes. We found that the phosphorylation of isoform-specific sites in eEF1A2-an essential translation elongation factor in neurons-is a key modulator of structural plasticity in dendritic spines. Expression of a nonphosphorylatable eEF1A2 mutant stimulated mRNA translation but reduced actin dynamics and spine density. By contrast, a phosphomimetic eEF1A2 mutant exhibited decreased association with F-actin and was inactive as a translation elongation factor. Activation of metabotropic glutamate receptor signaling triggered transient dissociation of eEF1A2 from its regulatory guanine exchange factor (GEF) protein in dendritic spines in a phosphorylation-dependent manner. We propose that eEF1A2 establishes a cross-talk mechanism that coordinates translation and actin dynamics during spine remodeling.

JTD Keywords: cytoskeleton, expression, f-actin, factor 1-alpha, factor 1a, messenger-rna, nucleotide exchange, protein-synthesis, synaptic plasticity, Aminoacyl-transfer-rna


Andreu, I, Falcones, B, Hurst, S, Chahare, N, Quiroga, X, Le Roux, AL, Kechagia, Z, Beedle, AEM, Elosegui-Artola, A, Trepat, X, Farre, R, Betz, T, Almendros, I, Roca-Cusachs, P, (2021). The force loading rate drives cell mechanosensing through both reinforcement and cytoskeletal softening Nature Communications 12, 4229

Cell response to force regulates essential processes in health and disease. However, the fundamental mechanical variables that cells sense and respond to remain unclear. Here we show that the rate of force application (loading rate) drives mechanosensing, as predicted by a molecular clutch model. By applying dynamic force regimes to cells through substrate stretching, optical tweezers, and atomic force microscopy, we find that increasing loading rates trigger talin-dependent mechanosensing, leading to adhesion growth and reinforcement, and YAP nuclear localization. However, above a given threshold the actin cytoskeleton softens, decreasing loading rates and preventing reinforcement. By stretching rat lungs in vivo, we show that a similar phenomenon may occur. Our results show that cell sensing of external forces and of passive mechanical parameters (like tissue stiffness) can be understood through the same mechanisms, driven by the properties under force of the mechanosensing molecules involved. Cells sense mechanical forces from their environment, but the precise mechanical variable sensed by cells is unclear. Here, the authors show that cells can sense the rate of force application, known as the loading rate, with effects on YAP nuclear localization and cytoskeletal stiffness remodelling.

JTD Keywords: Actin cytoskeleton, Actin filament, Actin-filament, Adhesion, Animal, Animals, Atomic force microscopy, Breathing, Cell, Cell adhesion, Cell culture, Cell nucleus, Cells, cultured, Cytoplasm, Extracellular-matrix, Fibroblast, Fibroblasts, Fibronectin, Frequency, Gene knockdown, Gene knockdown techniques, Genetics, Germfree animal, Integrin, Intracellular signaling peptides and proteins, Knockout mouse, Lung, Male, Mechanotransduction, Mechanotransduction, cellular, Metabolism, Mice, Mice, knockout, Microscopy, atomic force, Mouse, Optical tweezers, Paxillin, Physiology, Primary cell culture, Pxn protein, mouse, Rat, Rats, Rats, sprague-dawley, Respiration, Signal peptide, Softening, Specific pathogen-free organisms, Sprague dawley rat, Stress, Substrate, Substrate rigidity, Talin, Talin protein, mouse, Tln2 protein, mouse, Traction, Transmission, Ultrastructure, Yap1 protein, rat


Molina BG, del Valle LJ, Casanovas J, Lanzalaco S, Pérez-Madrigal MM, Turon P, Armelin E, Alemán C, (2021). Plasma-Functionalized Isotactic Polypropylene Assembled with Conducting Polymers for Bacterial Quantification by NADH Sensing Advanced Healthcare Materials 10, 2100425

Rapid detection of bacterial presence on implantable medical devices is essential to prevent biofilm formation, which consists of densely packed bacteria colonies able to withstand antibiotic-mediated killing. In this work, a smart approach is presented to integrate electrochemical sensors for detecting bacterial infections in biomedical implants made of isotactic polypropylene (i-PP) using chemical assembly. The electrochemical detection is based on the capacity of conducting polymers (CPs) to detect extracellular nicotinamide adenine dinucleotide (NADH) released from cellular respiration of bacteria, which allows distinguishing prokaryotic from eukaryotic cells. Oxygen plasma-functionalized free-standing i-PP, coated with a layer (≈1.1 µm in thickness) of CP nanoparticles obtained by oxidative polymerization, is used as working electrode for the anodic polymerization of a second CP layer (≈8.2 µm in thickness), which provides very high electrochemical activity and stability. The resulting layered material, i-PP /CP , detects the electro-oxidation of NADH in physiological media with a sensitivity 417 µA cm and a detection limit up to 0.14 × 10 m, which is below the concentration of extracellular NADH found for bacterial cultures of biofilm-positive and biofilm-negative strains. f 2 −2 −3

JTD Keywords: bacteria respiration, bacteria sensors, biomedical implants, flexible sensors, poly(3,4-ethylenedioxythiophene), Bacteria respiration, Bacteria sensors, Biomedical implants, Flexible sensors, Poly(3,4-ethylenedioxythiophene)


López-Canosa A, Perez-Amodio S, Yanac-Huertas E, Ordoño J, Rodriguez-Trujillo R, Samitier J, Castaño O, Engel E, (2021). A microphysiological system combining electrospun fibers and electrical stimulation for the maturation of highly anisotropic cardiac tissue Biofabrication 13, 35047

The creation of cardiac tissue models for preclinical testing is still a non-solved problem in drug discovery, due to the limitations related to thein vitroreplication of cardiac tissue complexity. Among these limitations, the difficulty of mimicking the functional properties of the myocardium due to the immaturity of the used cells hampers the obtention of reliable results that could be translated into human patients.In vivomodels are the current gold standard to test new treatments, although it is widely acknowledged that the used animals are unable to fully recapitulate human physiology, which often leads to failures during clinical trials. In the present work, we present a microfluidic platform that aims to provide a range of signaling cues to immature cardiac cells to drive them towards an adult phenotype. The device combines topographical electrospun nanofibers with electrical stimulation in a microfabricated system. We validated our platform using a co-culture of neonatal mouse cardiomyocytes and cardiac fibroblasts, showing that it allows us to control the degree of anisotropy of the cardiac tissue inside the microdevice in a cost-effective way. Moreover, a 3D computational model of the electrical field was created and validated to demonstrate that our platform is able to closely match the distribution obtained with the gold standard (planar electrode technology) using inexpensive rod-shaped biocompatible stainless-steel electrodes. The functionality of the electrical stimulation was shown to induce a higher expression of the tight junction protein Cx-43, as well as the upregulation of several key genes involved in conductive and structural cardiac properties. These results validate our platform as a powerful tool for the tissue engineering community due to its low cost, high imaging compatibility, versatility, and high-throughput configuration capabilities.

JTD Keywords: bioreactor, cardiac tissue engineering, cardiomyocytes, electrospinning, fabrication, fibroblasts, heart-on-a-chip, heart-tissue, in vitro models, myocardium, orientation, platform, scaffolds, Cardiac tissue engineering, Electrospinning, Field stimulation, Heart-on-a-chip, In vitro models, Microphysiological system


Parra-Monreal V, Ortega-Machuca MA, Ramon-Azcon J, Svendsen W, Romano-Rodriguez A, Moreno-Sereno M, (2021). Detection of cytokines in skeletal muscle tissue using optical SPR sensing platform Proceedings Of The 2021 13th Spanish Conference On Electron Devices, Cde 2021 , 102-105

In this work we have explored the use of a Surface Plasmon resonance (SPR) phenomenon for the detection of interleukin-6 (IL-6), a pro-inflammatory cytokine. It plays an important role in the muscle tissues, having direct relation with muscle contraction and, thus, it is considered a biomarker for some types of muscular dystrophies. Here we show that SPR can be used as a real-time monitoring of the shift of the reflectance dip of a gold diffraction grating in front to the antibody adhesion to gold.

JTD Keywords: Antibodies, Gratings, Interleukin-6 (il-6), Proteins, Surface plasmon resonance


Minguela J, Müller DW, Mücklich F, Llanes L, Ginebra MP, Roa JJ, Mas-Moruno C, (2021). Peptidic biofunctionalization of laser patterned dental zirconia: A biochemical-topographical approach Materials Science & Engineering C-Materials For Biological Applications 125, 112096

A dual approach employing peptidic biofunctionalization and laser micro-patterns on dental zirconia was explored, with the aim of providing a flexible tool to improve tissue integration of restorations. Direct laser interference patterning with a femtosecond Ti:Sapphire laser was employed, and two periodic grooved patterns were produced with a periodicity of 3 and 10 μm. A platform containing the cell-adhesive RGD and the osteogenic DWIVA peptides was used to functionalize the grooved surfaces. Topography and surface damage were characterized by confocal laser scanning (CLSM), scanning electron and scanning transmission electron microscopy techniques. The surface patterns exhibited a high homogeneity and subsurface damage was found in the form of nano-cracks and nano-pores, at the bottom of the valleys. Accelerated tests in water steam were carried out to assess hydrothermal degradation resistance, which slightly decreased after the laser treatment. Interestingly, the detrimental effects of the laser modification were reverted by a post-laser thermal treatment. The attachment of the molecule was verified trough fluorescence CLSM and X-ray photoelectron spectroscopy. Finally, the biological properties of the surfaces were studied in human mesenchymal stem cells. Cell adhesion, morphology, migration and differentiation were investigated. Cells on grooved surfaces displayed an elongated morphology and aligned along the patterns. On these surfaces, migration was greatly enhanced along the grooves, but also highly restricted in the perpendicular direction as compared to flat specimens. After biofunctionalization, cell number and cell area increased and well-developed cell cytoskeletons were observed. However, no effects on cell migration were found for the peptidic platform. Although some osteogenic potential was found in specimens grooved with a periodicity of 10 μm, the largest effects were observed from the biomolecule, which favored upregulation of several genes related to osteoblastic differentiation in all the surfaces.

JTD Keywords: alumina toughened zirconia, cell alignment, grain-size, implants, interference, laser patterning, osteogenic differentiation, osteointegration, peptides, surface functionalization, surface-topography, tissue, titanium surface, Laser patterning, Low-temperature degradation, Osteointegration, Peptides, Surface functionalization, Zirconia


Diaz-Lucena D, Kruse N, Thüne K, Schmitz M, Villar-Piqué A, da Cunha JEG, Hermann P, López-Pérez Ó, Andrés-Benito P, Ladogana A, Calero M, Vidal E, Riggert J, Pineau H, Sim V, Zetterberg H, Blennow K, del Río JA, Marín-Moreno A, Espinosa JC, Torres JM, Sánchez-Valle R, Mollenhauer B, Ferrer I, Zerr I, Llorens F, (2021). TREM2 expression in the brain and biological fluids in prion diseases Acta Neuropathologica 141, 841-859

Triggering receptor expressed on myeloid cells 2 (TREM2) is an innate immune cell surface receptor that regulates microglial function and is involved in the pathophysiology of several neurodegenerative diseases. Its soluble form (sTREM2) results from shedding of the TREM2 ectodomain. The role of TREM2 in prion diseases, a group of rapidly progressive dementias remains to be elucidated. In the present study, we analysed the expression of TREM2 and its main sheddase ADAM10 in the brain of sporadic Creutzfeldt-Jakob disease (sCJD) patients and evaluated the role of CSF and plasma sTREM2 as a potential diagnostic marker of prion disease. Our data indicate that, compared to controls, TREM2 is increased in sCJD patient brains at the mRNA and protein levels in a regional and subtype dependent fashion, and expressed in a subpopulation of microglia. In contrast, ADAM10 is increased at the protein, but not the mRNA level, with a restricted neuronal expression. Elevated CSF sTREM2 is found in sCJD, genetic CJD with mutations E200K and V210I in the prion protein gene (PRNP), and iatrogenic CJD, as compared to healthy controls (HC) (AUC = 0.78–0.90) and neurological controls (AUC = 0.73–0.85), while CSF sTREM2 is unchanged in fatal familial insomnia. sTREM2 in the CSF of cases with Alzheimer’s disease, and multiple sclerosis was not significantly altered in our series. CSF sTREM2 concentrations in sCJD are PRNP codon 129 and subtype-related, correlate with CSF 14-3-3 positivity, total-tau and YKL-40, and increase with disease progression. In plasma, sTREM2 is increased in sCJD compared with HC (AUC = 0.80), displaying positive correlations with plasma total-tau, neurofilament light, and YKL-40. We conclude that comparative study of TREM2 in brain and biological fluids of prion diseases reveals TREM2 to be altered in human prion diseases with a potential value in target engagement, patient stratification, and disease monitoring.

JTD Keywords: cerebrospinal fluid, creutzfeldt-jakob disease, microglia, plasma, prion diseases, Cerebrospinal fluid, Creutzfeldt-jakob disease, Microglia, Plasma, Prion diseases, Trem2


Fontana-Escartin A, Puiggalí-Jou A, Lanzalaco S, Bertran O, Alemán C, (2021). Manufactured Flexible Electrodes for Dopamine Detection: Integration of Conducting Polymer in 3D-Printed Polylactic Acid Advanced Engineering Materials 23, 2100002

Flexible electrochemical sensors based on electroactive materials have emerged as powerful analytical tools for biomedical applications requiring bioanalytes detection. Within this context, 3D printing is a remarkable technology for developing electrochemical devices, due to no design constraints, waste minimization, and batch manufacturing with high reproducibility. However, the fabrication of 3D printed electrodes is still limited by the in-house fabrication of conductive filaments, which requires the mixture of the electroactive material with melted of thermoplastic polymer (e.g., polylactic acid, PLA). Herein, a simple approach is presented for preparing electrochemical dopamine (DA) biosensors. Specifically, the surface of 3D-printed PLA specimens, which exhibit an elastic modulus and a tensile strength of 3.7 +/- 0.3 GPa and 47 +/- 1 MPa, respectively, is activated applying a 0.5 m NaOH solution for 30 min and, subsequently, poly(3,4-ethylenedioxythiophene) is polymerized in situ using aqueous solvent. The detection of DA with the produced sensors has been demonstrated by cyclic voltammetry, differential pulse voltammetry, and chronoamperometry. In summary, the obtained results reflect that low-cost electrochemical sensors, which are widely used in medicine and biotechnology, can be rapidly fabricated using the proposed approach that, although based on additive manufacturing, does not require the preparation of conductive filaments.

JTD Keywords: 3d printers, Additive manufacturing, Amines, Batch manufacturing, Biomedical applications, Chronoamperometry, Conducting polymer, Conducting polymers, Conductive filaments, Conservation, Cyclic voltammetry, Differential pulse voltammetry, Electroactive material, Electrochemical biosensor, Electrochemical devices, Electrochemical sensors, Electrodes, Electron emission, Flexible electrode, High reproducibility, Medical applications, Neurophysiology, Poly-3 ,4-ethylenedioxythiophene, Polyesters, Polylactic aci, Sodium hydroxide, Tensile strength, Thermoplastic polymer


Tornin J, Labay C, Tampieri F, Ginebra M-P, Canal C, (2021). Evaluation of the effects of cold atmospheric plasma and plasma-treated liquids in cancer cell cultures Nature Protocols 16, 2826-2850

Cold atmospheric plasma (CAP) is a potential anticancer therapy. CAP has cytotoxic effects when applied either directly to cancer cell cultures or indirectly through plasma-conditioned liquids. This protocol describes how to treat adherent cultures of human cancer cell lines with CAP or plasma-conditioned medium and determine cell viability following treatment. The protocol also includes details on how to quantify the reactive oxygen and nitrogen species present in medium following CAP treatment, using chemical probes using UV-visible or fluorescence spectroscopy. CAP treatment takes ~30 min, and 3 h are required to complete quantification of reactive oxygen and nitrogen species. By providing a standardized protocol for evaluation of the effects of CAP and plasma-conditioned medium, we hope to facilitate the comparison and interpretation of results seen across different laboratories. © 2021, The Author(s), under exclusive licence to Springer Nature Limited.

JTD Keywords: bacteria, decontamination, jet, skin, surface, Physical plasma


Torp N, Israelsen M, Madsen B, Lutz P, Jansen C, Strassburg C, Mortensen C, Knudsen AW, Sorensen GL, Holmskov U, Schlosser A, Thiele M, Trebicka J, Krag A, (2021). Level of MFAP4 in ascites independently predicts 1-year transplant-free survival in patients with cirrhosis Jhep Rep 3, 100287

Background & Aims: Prognostic models of cirrhosis underestimate disease severity for patients with cirrhosis and ascites. Microfibrillar-associated protein 4 (MFAP4) is an extracellular matrix protein linked to hepatic neoangiogenesis and fibrogenesis. We investigated ascites MFAP4 as a predictor of transplant-free survival in patients with cirrhosis and ascites. Methods: A dual-centre observational study of patients with cirrhosis and ascites recruited consecutively in relation to a paracentesis was carried out. Patients were followed up for 1 year, until death or liver transplantation (LTx). Ascites MFAP4 was tested with the model for end-stage liver disease (MELD-Na), CLIF Consortium Acute Decompensation (CLIF-C AD), and Child-Pugh score in Cox regression models. Results: Ninety-three patients requiring paracentesis were included. Median ascites MFAP4 was 29.7 U/L [22.3–41.3], and MELD-Na was 19 [16–23]. A low MELD-Na score (<20) was observed in 49 patients (53%). During follow-up, 20 patients died (22%), and 6 received LTx (6%). High ascites MFAP4 (>29.7 U/L) was associated with 1-year transplant-free survival (p = 0.002). In Cox regression, ascites MFAP4 and MELD-Na independently predicted 1-year transplant-free survival (hazard ratio [HR] = 0.97, p = 0.03, and HR = 1.08, p = 0.01, respectively). Ascites MFAP4 and CLIF-C AD also predicted survival independently (HR = 0.96, p = 0.02, and HR = 1.05, p = 0.03, respectively), whereas only ascites MFAP4 did, controlling for the Child-Pugh score (HR = 0.97, p = 0.03, and HR = 1.18, p = 0.16, respectively). For patients with MELD-Na <20, ascites MFAP4 but not ascites protein predicted 1-year transplant-free survival (HR 0.91, p = 0.02, and HR = 0.94, p = 0.17, respectively). Conclusions: Ascites MFAP4 predicts 1-year transplant-free survival in patients with cirrhosis and ascites. In patients with low MELD-Na scores, ascites MFAP4, but not total ascites protein, significantly predicted 1-year transplant-free survival. Lay summary: Patients with cirrhosis who have fluid in the abdomen, ascites, are at an increased risk of death and in need for liver transplantation. Our study identified patients with ascites and a poor prognosis by measuring microfibrillar associated protein 4 (MFAP4), a protein present in the abdominal fluid. Patients with low levels of the MFAP4 protein are at particularly increased risk of death or liver transplantation, suggesting that clinical care should be intensified in this group of patients. © 2021 The Authors

JTD Keywords: biomarker, clif-c ad, clif consortium acute decompensation, cps, child-pugh score, crp, c-reactive protein, ct, computed tomography, decompensated, ecm, extracellular matrix, fibrosis, fluid protein, gfr, glomerular filtration rate, hr, hazard ratio, inr, internationalised normal ratio, liver disease, liver-cirrhosis, ltx, liver transplantation, markers, meld-na, model for end-stage liver disease, mfap4, microfibrillar associated protein 4, mortality, nash, non-alcoholic steatohepatitis, natural-history, prognosis, risk-factors, sbp, spontaneous bacterial peritonitis, scores, stage, Biomarker, Decompensated, Egfr, estimated gfr, Fibrosis, Liver disease, Mortality, Prognosis, Spontaneous bacterial peritonitis


Ojosnegros, S, Seriola, A, Godeau, AL, Veiga, A, (2021). Embryo implantation in the laboratory: an update on current techniques Human Reproduction Update 27, 501-530

BACKGROUND: The embryo implantation process is crucial for the correct establishment and progress of pregnancy. During implantation, the blastocyst trophectoderm cells attach to the epithelium of the endometrium, triggering intense cell-to-cell crosstalk that leads to trophoblast outgrowth, invasion of the endometrial tissue, and formation of the placenta. However, this process, which is vital for embryo and foetal development in utero, is still elusive to experimentation because of its inaccessibility. Experimental implantation is cumbersome and impractical in adult animal models and is inconceivable in humans. OBJECTIVE AND RATIONALE: A number of custom experimental solutions have been proposed to recreate different stages of the implantation process in vitro, by combining a human embryo (or a human embryo surrogate) and endometrial cells (or a surrogate for the endometrial tissue). In vitro models allow rapid high-throughput interrogation of embryos and cells, and efficient screening of molecules, such as cytokines, drugs, or transcription factors, that control embryo implantation and the receptivity of the endometrium. However, the broad selection of available in vitro systems makes it complicated to decide which system best fits the needs of a specific experiment or scientific question. To orient the reader, this review will explore the experimental options proposed in the literature, and classify them into amenable categories based on the embryo/cell pairs employed. The goal is to give an overview of the tools available to study the complex process of human embryo implantation, and explain the differences between them, including the advantages and disadvantages of each system. SEARCH METHODS: We performed a comprehensive review of the literature to come up with different categories that mimic the different stages of embryo implantation in vitro, ranging from initial blastocyst apposition to later stages of trophoblast invasion or gastrulation. We will also review recent breakthrough advances on stem cells and organoids, assembling embryo-like structures and endometrial tissues. OUTCOMES: We highlight the most relevant systems and describe the most significant experiments. We focus on in vitro systems that have contributed to the study of human reproduction by discovering molecules that control implantation, including hormones, signalling molecules, transcription factors and cytokines. WIDER IMPLICATIONS: The momentum of this field is growing thanks to the use of stem cells to build embryo-like structures and endometrial tissues, and the use of bioengineering to extend the life of embryos in culture. We propose to merge bioengineering methods derived from the fields of stem cells and reproduction to develop new systems covering a wider window of the implantation process.

JTD Keywords: in vitro models, blastocyst, blastocyst-like structures, early-pregnancy, endometrial cells, epidermal-growth-factor, gene-expression, implantation, in vitro models, in-vitro model, indian hedgehog, organoids, receptivity, self-organization, spheroids, trophoblast, trophoblast invasion, uterine receptivity, Blastocyst, Blastocyst-like structures, Early-pregnancy, Endometrial cells, Endometrial stromal cells, Epidermal-growth-factor, Gene-expression, Implantation, In vitro models, In-vitro model, Indian hedgehog, Organoids, Receptivity, Self-organization, Spheroids, Trophoblast, Trophoblast invasion, Uterine receptivity


Dulay S, Rivas L, Miserere S, Pla L, Berdún S, Parra J, Eixarch E, Gratacós E, Illa M, Mir M, Samitier J, (2021). in vivo Monitoring with micro-implantable hypoxia sensor based on tissue acidosis Talanta 226, 122045

© 2020 Elsevier B.V. Hypoxia is a common medical problem, sometimes difficult to detect and caused by different situations. Control of hypoxia is of great medical importance and early detection is essential to prevent life threatening complications. However, the few current methods are invasive, expensive, and risky. Thus, the development of reliable and accurate sensors for the continuous monitoring of hypoxia is of vital importance for clinical monitoring. Herein, we report an implantable sensor to address these needs. The developed device is a low-cost, miniaturised implantable electrochemical sensor for monitoring hypoxia in tissue by means of pH detection. This technology is based on protonation/deprotonation of polypyrrole conductive polymer. The sensor was optimized in vitro and tested in vivo intramuscularly and ex vivo in blood in adult rabbits with respiration-induced hypoxia and correlated with the standard device ePOCTM. The sensor demonstrated excellent sensitivity and reproducibility; 46.4 ± 0.4 mV/pH in the pH range of 4–9 and the selectivity coefficient exhibited low interference activity in vitro. The device was linear (R2 = 0.925) with a low dispersion of the values (n = 11) with a cut-off of 7.1 for hypoxia in vivo and ex vivo. Statistics with one-way ANOVA (α = 0.05), shows statistical differences between hypoxia and normoxia states and the good performance of the pH sensor, which demonstrated good agreement with the standard device. The sensor was stable and functional after 18 months. The excellent results demonstrated the feasibility of the sensors in real-time monitoring of intramuscular tissue and blood for medical applications.

JTD Keywords: biocompatibility, blood-flow, clinical monitoring, electrochemical biosensor, electrodes, hypoxia, implantable sensor, in vivo tissue monitoring, ischemia, lactate, ph, ph sensor, rabbits, responses, vitro, Clinical monitoring, Dual signal outputs, Hypoxia, Implantable sensor, In vivo tissue monitoring, Ischemia, Ph sensor


Ortega MA, Rodríguez-Comas J, Yavas O, Velasco-Mallorquí F, Balaguer-Trias J, Parra V, Novials A, Servitja JM, Quidant R, Ramón-Azcón J, (2021). In Situ LSPR Sensing of Secreted Insulin in Organ-on-Chip Biosensors 11, 138

Organ-on-a-chip (OOC) devices offer new approaches for metabolic disease modeling and drug discovery by providing biologically relevant models of tissues and organs in vitro with a high degree of control over experimental variables for high-content screening applications. Yet, to fully exploit the potential of these platforms, there is a need to interface them with integrated non-labeled sensing modules, capable of monitoring, in situ, their biochemical response to external stimuli, such as stress or drugs. In order to meet this need, we aim here to develop an integrated technology based on coupling a localized surface plasmon resonance (LSPR) sensing module to an OOC device to monitor the insulin in situ secretion in pancreatic islets, a key physiological event that is usually perturbed in metabolic diseases such as type 2 diabetes (T2D). As a proof of concept, we developed a biomimetic islet-on-a-chip (IOC) device composed of mouse pancreatic islets hosted in a cellulose-based scaffold as a novel approach. The IOC was interfaced with a state-of-the-art on-chip LSPR sensing platform to monitor the in situ insulin secretion. The developed platform offers a powerful tool to enable the in situ response study of microtissues to external stimuli for applications such as a drug-screening platform for human models, bypassing animal testing.

JTD Keywords: biosensor, cytoarchitecture, dna hybridization, gelatin, in situ insulin monitoring, langerhans, lspr sensors, microfluidic device, organ-on-a-chip, parallel, platform, scaffold, Human pancreatic-islets, In situ insulin monitoring, Lspr sensors, Organ-on-a-chip


Guix M, Mestre R, Patiño T, de Corato M, Fuentes J, Zarpellon G, Sánchez S, (2021). Biohybrid soft robots with self-stimulating skeletons Science Robotics 6, eabe7577

Bioinspired hybrid soft robots that combine living and synthetic components are an emerging field in the development of advanced actuators and other robotic platforms (i.e., swimmers, crawlers, and walkers). The integration of biological components offers unique characteristics that artificial materials cannot precisely replicate, such as adaptability and response to external stimuli. Here, we present a skeletal muscle–based swimming biobot with a three-dimensional (3D)–printed serpentine spring skeleton that provides mechanical integrity and self-stimulation during the cell maturation process. The restoring force inherent to the spring system allows a dynamic skeleton compliance upon spontaneous muscle contraction, leading to a cyclic mechanical stimulation process that improves the muscle force output without external stimuli. Optimization of the 3D-printed skeletons is carried out by studying the geometrical stiffnesses of different designs via finite element analysis. Upon electrical actuation of the muscle tissue, two types of motion mechanisms are experimentally observed: directional swimming when the biobot is at the liquid-air interface and coasting motion when it is near the bottom surface. The integrated compliant skeleton provides both the mechanical self-stimulation and the required asymmetry for directional motion, displaying its maximum velocity at 5 hertz (800 micrometers per second, 3 body lengths per second). This skeletal muscle–based biohybrid swimmer attains speeds comparable with those of cardiac-based biohybrid robots and outperforms other muscle-based swimmers. The integration of serpentine-like structures in hybrid robotic systems allows self-stimulation processes that could lead to higher force outputs in current and future biomimetic robotic platforms. Copyright © 2021 The Authors, some rights reserved;

JTD Keywords: actuators, design, fabrication, mechanics, mems, myotubes, platform, tissue, 3d printers, Agricultural robots, Biological components, Biomimetic processes, Electrical actuation, Geometrical stiffness, Intelligent robots, Liquefied gases, Liquid-air interface, Mechanical integrity, Mechanical stimulation, Muscle, Muscle contractions, Phase interfaces, Robotics, Serpentine, Springs (components), Threedimensional (3-d)


de la Serna E, Arias-Alpízar K, Borgheti-Cardoso LN, Sanchez-Cano A, Sulleiro E, Zarzuela F, Bosch-Nicolau P, Salvador F, Molina I, Ramírez M, Fernàndez-Busquets X, Sánchez-Montalvá A, Baldrich E, (2021). Detection of Plasmodium falciparum malaria in 1 h using a simplified enzyme-linked immunosorbent assay Analytica Chimica Acta 1152, 338254

© 2021 Elsevier B.V. Malaria is a parasitic disease caused by protists of the genus Plasmodium, which are transmitted to humans through the bite of infected female Anopheles mosquitoes. Analytical methodologies and efficient drugs exist for the early detection and treatment of malaria, and yet this disease continues infecting millions of people and claiming several hundred thousand lives each year. One of the reasons behind this failure to control the disease is that the standard method for malaria diagnosis, microscopy, is time-consuming and requires trained personnel. Alternatively, rapid diagnostic tests, which have become common for point-of-care testing thanks to their simplicity of use, tend to be insufficiently sensitive and reliable, and PCR, which is sensitive, is too complex and expensive for massive population screening. In this work, we report a sensitive simplified ELISA for the quantitation of Plasmodium falciparum lactate dehydrogenase (Pf-LDH), which is capable of detecting malaria in 45–60 min. Assay development was founded in the selection of high-performance antibodies, implementation of a poly-horseradish peroxidase (polyHRP) signal amplifier, and optimization of whole-blood sample pre-treatment. The simplified ELISA achieved limits of detection (LOD) and quantification (LOQ) of 0.11 ng mL−1 and 0.37 ng mL−1, respectively, in lysed whole blood, and an LOD comparable to that of PCR in Plasmodium in vitro cultures (0.67 and 1.33 parasites μL−1 for ELISA and PCR, respectively). Accordingly, the developed immunoassay represents a simple and effective diagnostic tool for P. falciparum malaria, with a time-to-result of <60 min and sensitivity similar to the reference PCR, but easier to implement in low-resource settings.

JTD Keywords: malaria quantitative diagnosis, plasmodium culture, plasmodium ldh, polyhrp signal amplifier, simplified elisa, Malaria quantitative diagnosis, Plasmodium culture, Plasmodium ldh, Polyhrp signal amplifier, Simplified elisa


Castaño O, López-Mengual A, Reginensi D, Matamoros-Angles A, Engel E, del Rio JA, (2021). Chemotactic TEG3 Cells’ Guiding Platforms Based on PLA Fibers Functionalized With the SDF-1α/CXCL12 Chemokine for Neural Regeneration Therapy Frontiers In Bioengineering And Biotechnology 9, 627805

(Following spinal cord injury, olfactory ensheathing cell (OEC) transplantation is a promising therapeutic approach in promoting functional improvement. Some studies report that the migratory properties of OECs are compromised by inhibitory molecules and potentiated by chemical concentration differences. Here we compare the attachment, morphology, and directionality of an OEC-derived cell line, TEG3 cells, seeded on functionalized nanoscale meshes of Poly(l/dl-lactic acid; PLA) nanofibers. The size of the nanofibers has a strong effect on TEG3 cell adhesion and migration, with the PLA nanofibers having a 950 nm diameter being the ones that show the best results. TEG3 cells are capable of adopting a bipolar morphology on 950 nm fiber surfaces, as well as a highly dynamic behavior in migratory terms. Finally, we observe that functionalized nanofibers, with a chemical concentration increment of SDF-1α/CXCL12, strongly enhance the migratory characteristics of TEG3 cells over inhibitory substrates.

JTD Keywords: cell migration, cxcl12, electrospinning, gradients, pla nanofibers, sdf-1alpha, Cell migration, Cxcl12, Electrospinning, Gradients, Olfactory ensheathing cells, Pla nanofibers, Sdf-1alpha


Biosca A, Cabanach P, Abdulkarim M, Gumbleton M, Gómez-Canela C, Ramírez M, Bouzón-Arnáiz I, Avalos-Padilla Y, Borros S, Fernàndez-Busquets X, (2021). Zwitterionic self-assembled nanoparticles as carriers for Plasmodium targeting in malaria oral treatment Journal of Controlled Release 331, 364-375

© 2021 Elsevier B.V. The current decline in antimalarial drug efficacy due to the evolution of resistant Plasmodium strains calls for new strategies capable of improving the bioavailability of antimalarials, especially of those whose lipophilic character imparts them a low solubility in biological fluids. Here we have designed, synthesized and characterized amphiphilic zwitterionic block copolymers forming nanoparticles capable of penetrating the intestinal epithelium that can be used for oral administration. Poly(butyl methacrylate-co-morpholinoethyl sulfobetaine methacrylate) (PBMA-MESBMA)-based nanoparticles exhibited a specific targeting to Plasmodium falciparum-infected vs. parasite-free red blood cells (74.8%/0.8% respectively), which was maintained upon encapsulation of the lipophilic antimalarial drug curcumin (82.6%/0.3%). The in vitro efficacy of curcumin upon encapsulation was maintained relative to the free compound, with an IC50 around 5 μM. In vivo assays indicated a significantly increased curcumin concentration in the blood of mice one hour after being orally fed PBMA-MESBMA-curcumin in comparison to the administration of free drug (18.7 vs. 2.1 ng/ml, respectively). At longer times, however, plasma curcumin concentration equaled between free and encapsulated drug, which was reflected in similar in vivo antimalarial activities in Plasmodium yoelii yoelii-infected mice. Microscopic analysis in blood samples of fluorescently labeled PBMA-MESBMA revealed the presence of the polymer inside P. yoelii yoelii-parasitized erythrocytes one hour after oral administration to infected animals.

JTD Keywords: curcumin, drug delivery, malaria, pbma-mesbma, plasmodium, zwitterionic block copolymers, Curcumin, Drug delivery, Malaria, Pbma-mesbma, Plasmodium, Zwitterionic block copolymers


Fonte M, Tassi N, Fontinha D, Bouzón-Arnáiz I, Ferraz R, Araújo MJ, Fernàndez-Busquets X, Prudêncio M, Gomes P, Teixeira C, (2021). 4,9-Diaminoacridines and 4-Aminoacridines as Dual-Stage Antiplasmodial Hits Chemmedchem 16, 788-792

© 2020 Wiley-VCH GmbH Multi-stage drugs have been prioritized in antimalarial drug discovery, as targeting more than one process in the Plasmodium life cycle is likely to increase efficiency, while decreasing the chances of emergence of resistance by the parasite. Herein, we disclose two novel acridine-based families of compounds that combine the structural features of primaquine and chloroquine. Compounds prepared and studied thus far retained the in vitro activity displayed by the parent drugs against the erythrocytic stages of chloroquine-sensitive and -resistant Plasmodium falciparum strains, and against the hepatic stages of Plasmodium berghei, hence acting as dual-stage antiplasmodial hits.

JTD Keywords: acridines, antimalarial activity, blood-stage, liver-stage, malaria, plasmodium, Acridines, Antimalarial activity, Blood-stage, Liver-stage, Malaria, Plasmodium, Synthesis


Estrada-Petrocelli L, Torres A, Sarlabous L, Rafols-De-Urquia M, Ye-Lin Y, Prats-Boluda G, Jane R, Garcia-Casado J, (2021). Evaluation of Respiratory Muscle Activity by Means of Concentric Ring Electrodes Ieee Transactions On Biomedical Engineering 68, 1005-1014

© 1964-2012 IEEE. Surface electromyography (sEMG) can be used for the evaluation of respiratory muscle activity. Recording sEMG involves the use of surface electrodes in a bipolar configuration. However, electrocardiographic (ECG) interference and electrode orientation represent considerable drawbacks to bipolar acquisition. As an alternative, concentric ring electrodes (CREs) can be used for sEMG acquisition and offer great potential for the evaluation of respiratory muscle activity due to their enhanced spatial resolution and simple placement protocol, which does not depend on muscle fiber orientation. The aim of this work was to analyze the performance of CREs during respiratory sEMG acquisitions. Respiratory muscle sEMG was applied to the diaphragm and sternocleidomastoid muscles using a bipolar and a CRE configuration. Thirty-two subjects underwent four inspiratory load spontaneous breathing tests which was repeated after interchanging the electrode positions. We calculated parameters such as (1) spectral power and (2) median frequency during inspiration, and power ratios of inspiratory sEMG without ECG in relation to (3) basal sEMG without ECG (Rins/noise), (4) basal sEMG with ECG (Rins/cardio) and (5) expiratory sEMG without ECG (Rins/exp). Spectral power, Rins/noise and Rins/cardio increased with the inspiratory load. Significantly higher values (p < 0.05) of Rins/cardio and significantly higher median frequencies were obtained for CREs. Rins/noise and Rins/exp were higher for the bipolar configuration only in diaphragm sEMG recordings, whereas no significant differences were found in the sternocleidomastoid recordings. Our results suggest that the evaluation of respiratory muscle activity by means of sEMG can benefit from the remarkably reduced influence of cardiac activity, the enhanced detection of the shift in frequency content and the axial isotropy of CREs which facilitates its placement.

JTD Keywords: atmospheric measurements, concentric ring electrodes, electrocardiography, electrodes, electromyography, laplacian potential, non-invasive respiratory monitoring, particle measurements, respiratory muscles, surface electromyography, Concentric ring electrodes, Laplacian potential, Muscles, Non-invasive respiratory monitoring, Respiratory muscles, Surface electromyography


Seras-Franzoso J, Díaz-Riascos ZV, Corchero JL, González P, García-Aranda N, Mandaña M, Riera R, Boullosa A, Mancilla S, Grayston A, Moltó-Abad M, Garcia-Fruitós E, Mendoza R, Pintos-Morell G, Albertazzi L, Rosell A, Casas J, Villaverde A, Schwartz S, Abasolo I, (2021). Extracellular vesicles from recombinant cell factories improve the activity and efficacy of enzymes defective in lysosomal storage disorders Journal Of Extracellular Vesicles 10, e12058

In the present study the use of extracellular vesicles (EVs) as vehicles for therapeutic enzymes in lysosomal storage disorders was explored. EVs were isolated from mammalian cells overexpressing alpha-galactosidase A (GLA) or N-sulfoglucosamine sulfohydrolase (SGSH) enzymes, defective in Fabry and Sanfilippo A diseases, respectively. Direct purification of EVs from cell supernatants was found to be a simple and efficient method to obtain highly active GLA and SGSH proteins, even after EV lyophilization. Likewise, EVs carrying GLA (EV-GLA) were rapidly uptaken and reached the lysosomes in cellular models of Fabry disease, restoring lysosomal functionality much more efficiently than the recombinant enzyme in clinical use. In vivo, EVs were well tolerated and distributed among all main organs, including the brain. DiR-labelled EVs were localized in brain parenchyma 1 h after intra-arterial (internal carotid artery) or intravenous (tail vein) administrations. Moreover, a single intravenous administration of EV-GLA was able to reduce globotriaosylceramide (Gb3) substrate levels in clinically relevant tissues, such kidneys and brain. Overall, our results demonstrate that EVs from cells overexpressing lysosomal enzymes act as natural protein delivery systems, improving the activity and the efficacy of the recombinant proteins and facilitating their access to organs neglected by conventional enzyme replacement therapies.

JTD Keywords: alpha?galactosidase a, drug delivery, enzyme replacement therapy, fabry disease, lysosomal storage disorders, n-sulfoglucosamine sulfohydrolase, n?sulfoglucosamine sulfohydrolase, sanfilippo syndrome, Alpha-galactosidase a, Drug delivery, Enzyme replacement therapy, Fabry disease, Lysosomal storage disorders, N-sulfoglucosamine sulfohydrolase, Sanfilippo syndrome


Tornín J, Villasante A, Solé-Martí X, Ginebra MP, Canal C, (2021). Osteosarcoma tissue-engineered model challenges oxidative stress therapy revealing promoted cancer stem cell properties Free Radical Biology And Medicine 164, 107-118

© 2020 The Author(s) The use of oxidative stress generated by Cold Atmospheric Plasma (CAP) in oncology is being recently studied as a novel potential anti-cancer therapy. However, the beneficial effects of CAP for treating osteosarcoma have mostly been demonstrated in 2-dimensional cultures of cells, which do not mimic the complexity of the 3-dimensional (3D) bone microenvironment. In order to evaluate the effects of CAP in a relevant context of the human disease, we developed a 3D tissue-engineered model of osteosarcoma using a bone-like scaffold made of collagen type I and hydroxyapatite nanoparticles. Human osteosarcoma cells cultured within the scaffold showed a high capacity to infiltrate and proliferate and to exhibit osteomimicry in vitro. As expected, we observed significantly different functional behaviors between monolayer and 3D cultures when treated with Cold Plasma-Activated Ringer's Solution (PAR). Our data reveal that the 3D environment not only protects cells from PAR-induced lethality by scavenging and diminishing the amount of reactive oxygen and nitrogen species generated by CAP, but also favours the stemness phenotype of osteosarcoma cells. This is the first study that demonstrates the negative effect of PAR on cancer stem-like cell subpopulations in a 3D biomimetic model of cancer. These findings will allow to suitably re-focus research on plasma-based therapies in future.

JTD Keywords: 3d tumor model, cancer stem-like cells, cold atmospheric plasma, osteosarcoma, oxidative stress, plasma activated liquids, reactive oxygen and nitrogen species, 3d tumor model, Cancer stem-like cells, Cold atmospheric plasma, Osteosarcoma, Oxidative stress, Plasma activated liquids, Reactive oxygen and nitrogen species


Solé-Martí X, Espona-Noguera A, Ginebra MP, Canal C, (2021). Plasma-conditioned liquids as anticancer therapies In Vivo: Current state and future directions Cancers 13, 452

© 2021 by the authors. Licensee MDPI, Basel, Switzerland. During the last decade, cold atmospheric plasmas (CAP) have been broadly investigated for their therapeutic effect against cancer. CAP sources can be used to treat liquid media, thereby generating plasma-conditioned liquids (PCL). PCL represent a very interesting alternative to direct CAP treatment, because they may allow treatment of malignant tumors located in inner organs of the body by means of an injection, thus avoiding multiple surgeries. Although research on this therapy is still in its early stage, PCL have already demonstrated their potential anticancer effect in different types of cancer in vivo. This review gathers the existing literature involving PCL treatments in vivo, highlighting the differences between the approaches undertaken and the need for establishing standardized protocols in order to better understand the effects of PCL-based therapies in vivo. Plasma-conditioned liquids (PCL) are gaining increasing attention in the medical field, especially in oncology, and translation to the clinics is advancing on a good path. This emerging technology involving cold plasmas has great potential as a therapeutic approach in cancer diseases, as PCL have been shown to selectively kill cancer cells by triggering apoptotic mechanisms without damaging healthy cells. In this context, PCL can be injected near the tumor or intratumorally, thereby allowing the treatment of malignant tumors located in internal organs that are not accessible for direct cold atmospheric plasma (CAP) treatment. Therefore, PCL constitutes a very interesting and minimally invasive alternative to direct CAP treatment in cancer therapy, avoiding surgeries and allowing multiple local administrations. As the field advances, it is progressively moving to the evaluation of the therapeutic effects of PCL in in vivo scenarios. Exciting developments are pushing forward the clinical translation of this novel therapy. However, there is still room for research, as the quantification and identification of reactive oxygen and nitrogen species (RONS) in in vivo conditions is not yet clarified, dosage regimens are highly variable among studies, and other more relevant in vivo models could be used. In this context, this work aims to present a critical review of the state of the field of PCL as anticancer agents applied in in vivo studies.

JTD Keywords: cancer, cold atmospheric plasma, in vivo, Cancer, Cold atmospheric plasma, In vivo, Plasma-conditioned liquids


Fernández-Costa JM, Fernández-Garibay X, Velasco-Mallorquí F, Ramón-Azcón J, (2021). Bioengineered in vitro skeletal muscles as new tools for muscular dystrophies preclinical studies Journal Of Tissue Engineering 12, 2041731420981339

© The Author(s) 2021. Muscular dystrophies are a group of highly disabling disorders that share degenerative muscle weakness and wasting as common symptoms. To date, there is not an effective cure for these diseases. In the last years, bioengineered tissues have emerged as powerful tools for preclinical studies. In this review, we summarize the recent technological advances in skeletal muscle tissue engineering. We identify several ground-breaking techniques to fabricate in vitro bioartificial muscles. Accumulating evidence shows that scaffold-based tissue engineering provides topographical cues that enhance the viability and maturation of skeletal muscle. Functional bioartificial muscles have been developed using human myoblasts. These tissues accurately responded to electrical and biological stimulation. Moreover, advanced drug screening tools can be fabricated integrating these tissues in electrical stimulation platforms. However, more work introducing patient-derived cells and integrating these tissues in microdevices is needed to promote the clinical translation of bioengineered skeletal muscle as preclinical tools for muscular dystrophies.

JTD Keywords: biomaterials, drug screening platforms, muscular dystrophy, skeletal muscle, tissue engineering, Biomaterials, Drug screening platforms, Muscular dystrophy, Skeletal muscle, Tissue engineering


Mateu-Sanz, M, Tornin, J, Ginebra, MP, Canal, C, (2021). Cold Atmospheric Plasma: A New Strategy Based Primarily on Oxidative Stress for Osteosarcoma Therapy Journal Of Clinical Medicine 10, 893

Osteosarcoma is the most common primary bone tumor, and its first line of treatment presents a high failure rate. The 5-year survival for children and teenagers with osteosarcoma is 70% (if diagnosed before it has metastasized) or 20% (if spread at the time of diagnosis), stressing the need for novel therapies. Recently, cold atmospheric plasmas (ionized gases consisting of UV-Vis radiation, electromagnetic fields and a great variety of reactive species) and plasma-treated liquids have been shown to have the potential to selectively eliminate cancer cells in different tumors through an oxidative stress-dependent mechanism. In this work, we review the current state of the art in cold plasma therapy for osteosarcoma. Specifically, we emphasize the mechanisms unveiled thus far regarding the action of plasmas on osteosarcoma. Finally, we review current and potential future approaches, emphasizing the most critical challenges for the development of osteosarcoma therapies based on this emerging technique.

JTD Keywords: cancer stem cells, cold atmospheric plasma, osteosarcoma, oxidative stress, plasma treated liquids, reactive oxygen and nitrogen species, Antineoplastic activity, Antineoplastic agent, Cancer chemotherapy, Cancer stem cell, Cancer stem cells, Cancer surgery, Cancer survival, Cell therapy, Cold atmospheric plasma, Cold atmospheric plasma therapy, Electromagnetism, Human, In vitro study, Intracellular signaling, Oncogene, Osteosarcoma, Oxidative stress, Plasma treated liquids, Reactive nitrogen species, Reactive oxygen and nitrogen species, Reactive oxygen metabolite, Review, Tumor microenvironment


Pinheiro, ND, Freire, RT, Conrado, JAM, Batista, AD, Petruci, JFD, (2021). Paper-based optoelectronic nose for identification of indoor air pollution caused by 3D printing thermoplastic filaments Analytica Chimica Acta 1143, 1-8

Commercial printers based on fused deposition modeling (FDM) are widely adopted for 3D printing applications. This method consists of the heating of polymeric filaments over the melting point followed by their deposition onto a solid base to create the desirable 3D structure. Prior investigation using chromatographic techniques has shown that chemical compounds (e.g. VOCs), which can be harmful to users, are emitted during the printing process, producing adverse effects to human health and contributing to indoor air pollution. In this study, we present a simple, inexpensive and disposable paperbased optoelectronic nose (i.e. colorimetric sensor array) to identify the gaseous emission fingerprint of five different types of thermoplastic filaments (ABS, TPU, PETG, TRITAN and PLA) in the indoor environment. The optoelectronic nose is comprised of selected 15 dyes with different chemical properties deposited onto a microfluidic paper-based device with spots of 5 mm in diameter each. Digital images were obtained from an ordinary flatbed scanner, and the RGB information collected before and after air exposure was extracted by using an automated routine designed in MATLAB, in which the color changes provide a unique fingerprint for each filament in 5 min of printing. Reproducibility was obtained in the range of 2.5-10% (RSD). Hierarchical clustering analysis (HCA) and principal component analysis (PCA) were successfully employed, showing suitable discrimination of all studied filaments and the non-polluted air. Besides, air spiked with vapors of the most representative VOCs were analyzed by the optoelectronic nose and visually compared to each filament. The described study shows the potential of the paper-based optoelectronic nose to monitor possible hazard emissions from 3D printers. (C) 2020 Elsevier B.V. All rights reserved.

JTD Keywords: 3d printing, colorimetric sensor array, indoor air pollution, optoelectronic nose, paper-based, 3d printing, Colorimetric sensor array, Emissions, Indoor air pollution, Optoelectronic nose, Paper-based, Thermoplastic filaments


Vidal, E, Guillem-Marti, J, Ginebra, MP, Combes, C, Ruperez, E, Rodriguez, D, (2021). Multifunctional homogeneous calcium phosphate coatings: Toward antibacterial and cell adhesive titanium scaffolds Surface & Coatings Technology 405, 126557

Implants for orthopedic applications need to be biocompatible and bioactive, with mechanical properties similar to those of surrounding natural bone. Given this scenario titanium (Ti) scaffolds obtained by Direct Ink Writing technique offer the opportunity to manufacture customized structures with controlled porosity and mechanical properties. Considering that 3D Ti scaffolds have a significant surface area, it is necessary to develop strategies against the initial bacterial adhesion in order to prevent infection in the early stages of the implantation, while promoting cell adhesion to the scaffold. The challenge is not only achieving a balance between antibacterial activity and osseointegration, it is also to develop a homogeneous coating on the inner and outer surface of the scaffold. The purpose of this work was the development of a single-step electrodeposition process in order to uniformly cover Ti scaffolds with a layer of calcium phosphate (CaP) loaded with chlorhexidine digluconate (CHX). Scaffold characterization was assessed by scanning electron microscopy, Energy dispersive X-ray spectroscopy, X-ray diffraction, micro-Raman microscopy and compressive strength tests. Results determined that the surface of scaffolds was covered by plate-like and whisker-like calcium phosphate crystals, which main phases were octacalcium phosphate and brushite. Biological tests showed that the as-coated scaffolds reduced bacteria adhesion (73 +/- 3% for Staphylococcus aureus and 70 +/- 2% for Escherichia coli). In vitro cell studies and confocal analysis revealed the adhesion and spreading of osteoblast-like SaOS-2 on coated surfaces. Therefore, the proposed strategy can be a potential candidate in bone replacing surgeries.

JTD Keywords: Antibacterial, Bacterial, Behavior, Biocompatibility, Calcium phosphate coating, Chlorhexidine, Chlorhexidine digluconate, Deposition, Electrodeposition, Hydroxyapatite coatings, Implants, One-step pulse electrodeposition, Plasma-spray, Release, Surface, Titanium scaffolds


Mateu-Sanz, M., Tornín, J., Brulin, B., Khlyustova, A., Ginebra, M. P., Layrolle, P., Canal, C., (2020). Cold plasma-treated ringer's saline: A weapon to target osteosarcoma Cancers 12, (1), 227

Osteosarcoma (OS) is the main primary bone cancer, presenting poor prognosis and difficult treatment. An innovative therapy may be found in cold plasmas, which show anti-cancer effects related to the generation of reactive oxygen and nitrogen species in liquids. In vitro models are based on the effects of plasma-treated culture media on cell cultures. However, effects of plasma-activated saline solutions with clinical application have not yet been explored in OS. The aim of this study is to obtain mechanistic insights on the action of plasma-activated Ringer’s saline (PAR) for OS therapy in cell and organotypic cultures. To that aim, cold atmospheric plasma jets were used to obtain PAR, which produced cytotoxic effects in human OS cells (SaOS-2, MG-63, and U2-OS), related to the increasing concentration of reactive oxygen and nitrogen species generated. Proof of selectivity was found in the sustained viability of hBM-MSCs with the same treatments. Organotypic cultures of murine OS confirmed the time-dependent cytotoxicity observed in 2D. Histological analysis showed a decrease in proliferating cells (lower Ki-67 expression). It is shown that the selectivity of PAR is highly dependent on the concentrations of reactive species, being the differential intracellular reactive oxygen species increase and DNA damage between OS cells and hBM-MSCs key mediators for cell apoptosis.

JTD Keywords: Bone cancer, Cold atmospheric plasma, Organotypic model, Osteosarcoma, Plasma-activated liquid, Reactive species, Ringer's saline


Rodríguez-Contreras, A., Torres, D., Guillem-Marti, J., Sereno, P., Ginebra, M. P., Calero, J. A., Manero, J. M., Rupérez, E., (2020). Development of novel dual-action coatings with osteoinductive and antibacterial properties for 3D-printed titanium implants Surface and Coatings Technology 403, 126381

Gallium (Ga) has been recently proposed as a novel therapeutic agent, since it promotes bone formation and exhibits antibacterial properties. This work focuses on the optimization of a thermochemical treatment that incorporates Ga ions by the addition of the body-friendly Ga nitrate approved by the Food and Drug Administration. The objective was to simultaneously provide the inner and the outer surfaces of porous‑titanium surfaces obtained by 3D-printing with bioactivity and antibacterial properties. The apatite-forming ability of the coating, as well as the antibacterial activity and SaOS-2 cell adhesion, proliferation, differentiation and mineralization were evaluated and compared with untreated Ti surfaces. The characterization of the surfaces revealed the presence of a Ga-containing calcium titanate layer, which was non cytotoxic and in simulated body fluid produced a homogeneous apatite coating well adhered to the substrate. The formation of this apatite layer was accelerated with increasing Ga amounts present on the surface, resulting also in an increase in thickness. An initial quick release of Ga ion promoted the antibacterial effect against gram positive strains, especially for Pseudomonas aeruginosa, one of the most frequent resistant pathogens in nosocomial infections. SaOS-2 cells adhered and proliferated on the Ga-doped Ti surfaces, its presence contributed to cell differentiation and to considerably increase the mineralization levels. Thus, the developed multifunctional coatings could provide bioactivity to the porous Ti implants while protecting them from the most frequent gram-negative pathogens.

JTD Keywords: 3D-printing, Antibacterial activity, Biomaterials, Gallium, Porous structures, Titanium implants


Lopez-Muñoz, Gerardo A., Ortega, Maria Alejandra, Ferret-Miñana, Ainhoa, De Chiara, Francesco, Ramón-Azcón, Javier, (2020). Direct and label-free monitoring of albumin in 2D fatty liver disease model using plasmonic nanogratings Nanomaterials 10, (12), 2520

Non-alcoholic fatty liver (NAFLD) is a metabolic disorder related to a chronic lipid accumulation within the hepatocytes. This disease is the most common liver disorder worldwide, and it is estimated that it is present in up to 25% of the world’s population. However, the real prevalence of this disease and the associated disorders is unknown mainly because reliable and applicable diagnostic tools are lacking. It is known that the level of albumin, a pleiotropic protein synthesized by hepatocytes, is correlated with the correct function of the liver. The development of a complementary tool that allows direct, sensitive, and label-free monitoring of albumin secretion in hepatocyte cell culture can provide insight into NAFLD’s mechanism and drug action. With this aim, we have developed a simple integrated plasmonic biosensor based on gold nanogratings from periodic nanostructures present in commercial Blu-ray optical discs. This sensor allows the direct and label-free monitoring of albumin in a 2D fatty liver disease model under flow conditions using a highly-specific polyclonal antibody. This technology avoids both the amplification and blocking steps showing a limit of detection within pM range (≈0.26 ng/mL). Thanks to this technology, we identified the optimal fetal bovine serum (FBS) concentration to maximize the cells’ lipid accumulation. Moreover, we discovered that the hepatocytes increased the amount of albumin secreted on the third day from the lipids challenge. These data demonstrate the ability of hepatocytes to respond to the lipid stimulation releasing more albumin. Further investigation is needed to unveil the biological significance of that cell behavior.

JTD Keywords: 2D fatty liver in vitro model, Blu-Ray disc, Plasmonic nanomaterials, Label-Free Biosensing


Ruiz-Vega, G., Arias-Alpízar, K., de la Serna, E., Borgheti-Cardoso, L. N., Sulleiro, E., Molina, I., Fernàndez-Busquets, X., Sánchez-Montalvá, A., del Campo, F. J., Baldrich, E., (2020). Electrochemical POC device for fast malaria quantitative diagnosis in whole blood by using magnetic beads, Poly-HRP and microfluidic paper electrodes Biosensors and Bioelectronics 150, 111925

Malaria, a parasitic infection caused by Plasmodium parasites and transmitted through the bite of infected female Anopheles mosquitos, is one of the main causes of mortality in many developing countries. Over 200 million new infections and nearly half a million deaths are reported each year, and more than three billion people are at risk of acquiring malaria worldwide. Nevertheless, most malaria cases could be cured if detected early. Malaria eradication is a top priority of the World Health Organisation. However, achieving this goal will require mass population screening and treatment, which will be hard to accomplish with current diagnostic tools. We report an electrochemical point-of-care device for the fast, simple and quantitative detection of Plasmodium falciparum lactate dehydrogenase (PfLDH) in whole blood samples. Sample analysis includes 5-min lysis to release intracellular parasites, and stirring for 5 more min with immuno-modified magnetic beads (MB) along with an immuno-modified signal amplifier. The rest of the magneto-immunoassay, including sample filtration, MB washing and electrochemical detection, is performed at a disposable paper electrode microfluidic device. The sensor provides PfLDH quantitation down to 2.47 ng mL−1 in spiked samples and for 0.006–1.5% parasitemias in Plasmodium-infected cultured red blood cells, and discrimination between healthy individuals and malaria patients presenting parasitemias >0.3%. Quantitative malaria diagnosis is attained with little user intervention, which is not achieved by other diagnostic methods.

JTD Keywords: Electrochemical magneto-immunosensor, Malaria quantitative diagnosis, Paper microfluidic electrode, Plasmodium LDH, Point-of-care (POC) testing


Hoogduijn, M.J., Montserrat, N., van der Laan, L.J.W., Dazzi, F., Perico, N., Kastrup, J., Gilbo, N., Ploeg, R.J., Roobrouck, V., Casiraghi, F., Johnson, C.L., Franquesa, M., Dahlke, M.H., Massey, E., Hosgood, S., Reinders, M.E.J., (2020). The emergence of regenerative medicine in organ transplantation: 1st European Cell Therapy and Organ Regeneration Section meeting Transplant International 33, (8), 833-840

Regenerative medicine is emerging as a novel field in organ transplantation. In September 2019, the European Cell Therapy and Organ Regeneration Section (ECTORS) of the European Society for Organ Transplantation (ESOT) held its first meeting to discuss the state-of-the-art of regenerative medicine in organ transplantation. The present article highlights the key areas of interest and major advances in this multidisciplinary field in organ regeneration and discusses its implications for the future of organ transplantation.

JTD Keywords: Cell therapy, Machine perfusion, Mesenchymal stromal cell, Organoid, Regeneration, Transplantation


Labay, C., Roldán, M., Tampieri, F., Stancampiano, A., Bocanegra, P. E., Ginebra, M. P., Canal, C., (2020). Enhanced generation of reactive species by cold plasma in gelatin solutions for selective cancer cell death ACS Applied Materials and Interfaces 12, (42), 47256-47269

Atmospheric pressure plasma jets generate reactive oxygen and nitrogen species (RONS) in liquids and biological media, which find application in the new area of plasma medicine. These plasma-treated liquids were demonstrated recently to possess selective properties on killing cancer cells and attracted attention toward new plasma-based cancer therapies. These allow for local delivery by injection in the tumor but can be quickly washed away by body fluids. By confining these RONS in a suitable biocompatible delivery system, great perspectives can be opened in the design of novel biomaterials aimed for cancer therapies. Gelatin solutions are evaluated here to store RONS generated by atmospheric pressure plasma jets, and their release properties are evaluated. The concentration of RONS was studied in 2% gelatin as a function of different plasma parameters (treatment time, nozzle distance, and gas flow) with two different plasma jets. Much higher production of reactive species (H2O2 and NO2-) was revealed in the polymer solution than in water after plasma treatment. The amount of RONS generated in gelatin is greatly improved with respect to water, with concentrations of H2O2 and NO2- between 2 and 12 times higher for the longest plasma treatments. Plasma-treated gelatin exhibited the release of these RONS to a liquid media, which induced an effective killing of bone cancer cells. Indeed, in vitro studies on the sarcoma osteogenic (SaOS-2) cell line exposed to plasma-treated gelatin led to time-dependent increasing cytotoxicity with the longer plasma treatment time of gelatin. While the SaOS-2 cell viability decreased to 12%-23% after 72 h for cells exposed to 3 min of treated gelatin, the viability of healthy cells (hMSC) was preserved (?90%), establishing the selectivity of the plasma-treated gelatin on cancer cells. This sets the basis for designing improved hydrogels with high capacity to deliver RONS locally to tumors.

JTD Keywords: Cold atmospheric plasma, Hydrogel, Osteosarcoma, Reactive oxygen and nitrogen species


Borgheti-Cardoso, L. N., Kooijmans, S. A. A., Gutiérrez Chamorro, L., Biosca, A., Lantero, E., Ramírez, M., Avalos-Padilla, Y., Crespo, I., Fernández, I., Fernandez-Becerra, C., del Portillo, H. A., Fernàndez-Busquets, X., (2020). Extracellular vesicles derived from Plasmodium-infected and non-infected red blood cells as targeted drug delivery vehicles International Journal of Pharmaceutics 587, 119627

Among several factors behind drug resistance evolution in malaria is the challenge of administering overall doses that are not toxic for the patient but that, locally, are sufficiently high to rapidly kill the parasites. Thus, a crucial antimalarial strategy is the development of drug delivery systems capable of targeting antimalarial compounds to Plasmodium with high specificity. In the present study, extracellular vesicles (EVs) have been evaluated as a drug delivery system for the treatment of malaria. EVs derived from naive red blood cells (RBCs) and from Plasmodium falciparum-infected RBCs (pRBCs) were isolated by ultrafiltration followed by size exclusion chromatography. Lipidomic characterization showed that there were no significant qualitative differences between the lipidomic profiles of pRBC-derived EVs (pRBC-EVs) and RBC-derived EVs (RBC-EVs). Both EVs were taken up by RBCs and pRBCs, although pRBC-EVs were more efficiently internalized than RBC-EVs, which suggested their potential use as drug delivery vehicles for these cells. When loaded into pRBC-EVs, the antimalarial drugs atovaquone and tafenoquine inhibited in vitro P. falciparum growth more efficiently than their free drug counterparts, indicating that pRBC-EVs can potentially increase the efficacy of several small hydrophobic drugs used for the treatment of malaria.

JTD Keywords: Antimalarial drugs, Drug delivery, Extracellular vesicles, Malaria, Plasmodium falciparum


Lantero, E., Fernandes, J., Aláez-Versón, C. R., Gomes, J., Silveira, H., Nogueira, F., Fernàndez-Busquets, X., (2020). Heparin administered to anopheles in membrane feeding assays blocks plasmodium development in the mosquito Biomolecules 10, (8), 1136

Innovative antimalarial strategies are urgently needed given the alarming evolution of resistance to every single drug developed against Plasmodium parasites. The sulfated glycosaminoglycan heparin has been delivered in membrane feeding assays together with Plasmodium berghei-infected blood to Anopheles stephensi mosquitoes. The transition between ookinete and oocyst pathogen stages in the mosquito has been studied in vivo through oocyst counting in dissected insect midguts, whereas ookinete interactions with heparin have been followed ex vivo by flow cytometry. Heparin interferes with the parasite’s ookinete–oocyst transition by binding ookinetes, but it does not affect fertilization. Hypersulfated heparin is a more efficient blocker of ookinete development than native heparin, significantly reducing the number of oocysts per midgut when offered to mosquitoes at 5 µg/mL in membrane feeding assays. Direct delivery of heparin to mosquitoes might represent a new antimalarial strategy of rapid implementation, since it would not require clinical trials for its immediate deployment.

JTD Keywords: Anopheles, Antimalarial drugs, Heparin, Malaria, Mosquito, Ookinete, Plasmodium, Transmission blocking


Majchrowicz, A., Roguska, A., Krawczy, Lewandowska, M., Martí-Muñoz, J., Engel, E., Castano, O., (2020). In vitro evaluation of degradable electrospun polylactic acid/bioactive calcium phosphate ormoglass scaffolds Archives of Civil and Mechanical Engineering 20, (2), 50

Nowadays, the main limitation for clinical application of scaffolds is considered to be an insufficient vascularization of the implanted platforms and healing tissues. In our studies, we proposed a novel PLA-based hybrid platform with aligned and random fibrous internal structure and incorporated calcium phosphate (CaP) ormoglass nanoparticles (0, 10, 20 and 30 wt%) as an off-the-shelf method for obtaining scaffolds with pro-angiogenic properties. Complex morphological and physicochemical evaluation of PLA–CaP ormoglass composites was performed before and after in vitro degradation test in SBF solution to assess their biological potential. The degradation process of PLA–CaP ormoglass composites was accompanied by numerous CaP-based precipitations with extended topography and cauliflower-like shape which may enhance bonding of the material with the bone tissue and accelerate the regenerative process. Random fiber orientation was preferable for CaP compounds deposition upon in vitro degradation. CaP compounds precipitated firstly for randomly oriented composite nonwovens with 20 and 30 wt% addition of ormoglass. Moreover, the preliminary bioactivity test has shown that BSA adsorbed to PLA–CaP ormoglass composites (both aligned and randomly oriented) with 20 and 30 wt% of ormoglass nanoparticles which was not observed for pure PLA scaffolds.

JTD Keywords: Calcium phosphate ormoglass, Composites, Degradation, Electrospinning, PLA


Hamouda, I., Labay, C., Ginebra, M. P., Nicol, E., Canal, C., (2020). Investigating the atmospheric pressure plasma jet modification of a photo-crosslinkable hydrogel Polymer 192, 122308

Atmospheric pressure plasma jets (APPJ) have great potential in wound healing, bacterial disinfection and in cancer therapy. Recent studies pointed out that hydrogels can be used as screens during APPJ treatment, or even be used as reservoirs for reactive oxygen and nitrogen species generated by APPJ in liquids. Thus, novel applications are emerging for hydrogels which deserve fundamental exploration of the possible modifications undergone by the polymers in solution due to the reactivity with plasmas. Here we investigate the possible modifications occurred by APPJ treatment of an amphiphilic poly(ethylene oxide)-based triblock copolymer (tPEO) photo-crosslinkable hydrogel. While APPJ treatments lead to a certain degradation of the self-assembly of the polymeric chains at low concentrations (<2 g/L), at the higher concentrations required to form a hydrogel (>2 g/L), the polymeric chains are unaffected by APPJ and the hydrogel forming ability is kept. APPJ treatments induced a pre-crosslinking of the network with an increase of the mechanical properties of the hydrogel. Overall, the small modifications induced allow thinking of polymer solutions with hydrogel forming ability a new platform for several applications related to plasma medicine, and thus, with potential in different therapies.

JTD Keywords: Atmospheric pressure plasma jet, Hydrogel, Photo-crosslinking, Polymer solution, Self-assembly


Duran, Jordi, Brewer, M. Kathryn, Hervera, Arnau, Gruart, Agnès, del Rio, Jose Antonio, Delgado-García, José M., Guinovart, Joan J., (2020). Lack of astrocytic glycogen alters synaptic plasticity but not seizure susceptibility Molecular Neurobiology 57, 4657–4666

Brain glycogen is mainly stored in astrocytes. However, recent studies both in vitro and in vivo indicate that glycogen also plays important roles in neurons. By conditional deletion of glycogen synthase (GYS1), we previously developed a mouse model entirely devoid of glycogen in the central nervous system (GYS1Nestin-KO). These mice displayed altered electrophysiological properties in the hippocampus and increased susceptibility to kainate-induced seizures. To understand which of these functions are related to astrocytic glycogen, in the present study, we generated a mouse model in which glycogen synthesis is eliminated specifically in astrocytes (GYS1Gfap-KO). Electrophysiological recordings of awake behaving mice revealed alterations in input/output curves and impaired long-term potentiation, similar, but to a lesser extent, to those obtained with GYS1Nestin-KO mice. Surprisingly, GYS1Gfap-KO mice displayed no change in susceptibility to kainate-induced seizures as determined by fEPSP recordings and video monitoring. These results confirm the importance of astrocytic glycogen in synaptic plasticity.

JTD Keywords: Astrocyte, Epilepsy, Glycogen, Long-term potentiation, Metabolism, Plasticity.


Rivas, L., Dulay, S., Miserere, S., Pla, L., Marin, S. B., Parra, J., Eixarch, E., Gratacós, E., Illa, M., Mir, M., Samitier, J., (2020). Micro-needle implantable electrochemical oxygen sensor: ex-vivo and in-vivo studies Biosensors and Bioelectronics 153, 112028

Oxygen is vital for energy metabolism in mammals and the variability of the concentration is considered a clinical alert for a wide range of metabolic malfunctions in medicine. In this article, we describe the development and application of a micro-needle implantable platinum-based electrochemical sensor for measuring partial pressure of oxygen in intramuscular tissue (in-vivo) and vascular blood (ex-vivo). The Pt-Nafion® sensor was characterized morphological and electrochemically showing a higher sensitivity of −2.496 nA/mmHg (−1.495 nA/μM) when comparing with its bare counterpart. Our sensor was able to discriminate states with different oxygen partial pressures (pO2) for ex-vivo (blood) following the same trend of the commercial gas analyzer used as standard. For in-vivo (intramuscular) experiments, since there is not a gold standard for measuring pO2 in tissue, it was not possible to correlate the obtained currents with the pO2 in tissue. However, our sensor was able to detect clear statistical differences of O2 between hyperoxia and hypoxia states in tissue.

JTD Keywords: Hypoxia, Implantable sensor, In-vivo test, Ischemia, Nafion, Oxygen sensor


Lanzalaco, S., Fabregat, G., Muñoz-Galan, H., Cabrera, J., Muñoz-Pascual, X., Llorca, J., Alemán, C., (2020). Recycled low-density polyethylene for noninvasive glucose monitoring: A proposal for plastic recycling that adds technological value ACS Sustainable Chemistry and Engineering 8, (33), 12554-12560

In this work, we present a successful strategy to convert recycled LDPE films, which usually end up in landfills or leak into the environment, into an advanced biomedical product. More specifically, LDPE films for food packaging have been treated with atmosphere corona discharge plasma for electrochemical detection of glucose. Enzyme-functionalized sensors manufactured using such recycled materials, which act as a mediator capable of electrocommunicating with the glucose oxidase (GOx) enzyme, are able to detect glucose concentrations in sweat and are fully compatible with the levels of such bioanalytes in both healthy and diabetic patients. Covalent immobilization of the GOx enzyme on the plasma-treated LDPE films has been successfully performed using the carbodiimide method, as proved by X-ray photoelectron spectroscopy. Then, the electronic communication between the deeply buried active site of the GOx and the reactive excited species formed at the surface of the plasma-treated LDPE has been demonstrated by linear sweep voltammetry. Finally, cyclic voltammetry in artificial sweat has been used to show that the LDPE-functionalized sensor has a linear response in the concentration of range of 50 μM to 1 mM with a limit of detection of 375 μA·μM–1·cm–2. Comparison of the performance of sensors prepared using recycled (i.e. with additives) and pristine (i.e. without additives) LDPE indicates that the utilization of the former does not require any pretreatment to eliminate additives. The present strategy demonstrates a facile approach for recycling LDPE waste into a high value-added product, which will potentially pave the way for the treatment of plastic waste in the future. Noninvasive glucose sensors based on recycled LDPE may play a crucial role in monitoring diabetes in underdeveloped regions.

JTD Keywords: Biosensors, Diabetes monitoring, High-value recycling, Plasma treatment, Sweat sensors


Lanzalaco, S., Turon, P., Weis, C., Mata, C., Planas, E., Alemán, C., Armelin, E., (2020). Toward the new generation of surgical meshes with 4D response: Soft, dynamic, and adaptable Advanced Functional Materials 30, (36), 2004145

Herein, a facile approach toward transforming a 2D polypropylene flexible mesh material into a 4D dynamic system is presented. The versatile platform, composed by a substrate of knitted fibers of isotactic polypropylene (iPP) mesh and a coating of thermosensitive poly(N‐isopropylacrylamide‐co‐N,N’‐methylene bis(acrylamide) (PNIPAAm‐co‐MBA) hydrogel, covalently bonded to the mesh surface, after cold‐plasma surface treatment and radical polymerization, is intended to undergo variations in its geometry via its reversible folding/unfolding behavior. The study is the first to trace the 3D movement of a flat surgical mesh, intended to repair hernia defects, under temperature and humidity control. An infrared thermographic camera and an optical microscope are used to evaluate the macroscopic and microscopic structure stimulus response. The presence of the PP substrate and the distribution of the gel surrounding the PP threads, affect both the PNIPAAM gel expansion/contraction as well as the time of folding/unfolding response. Furthermore, PP‐g‐PNIPAAm meshes show an increase in the bursting strength of ≈16% with respect to the uncoated mesh, offering a strongest and adaptable system for its future implantation in human body. The findings reported offer unprecedented application possibilities in the biomedical field.

JTD Keywords: Dynamic devices, Polypropylene meshes, Surgical implants, Thermosensitive hydrogels


Cilloni, Daniela, Petiti, Jessica, Campia, Valentina, Podestà , Marina, Squillario, Margherita, Montserrat, Nuria, Bertaina, Alice, Sabatini, Federica, Carturan, Sonia, Berger, Massimo, Saglio, Francesco, Bandini, Giuseppe, Bonifazi, Francesca, Fagioli, Franca, Moretta, Lorenzo, Saglio, Giuseppe, Verri, Alessandro, Barla, Annalisa, Locatelli, Franco, Frassoni, Francesco, (2020). Transplantation induces profound changes in the transcriptional asset of hematopoietic stem cells: Identification of specific signatures using machine learning techniques Journal of Clinical Medicine 9, (6), 1670

During the phase of proliferation needed for hematopoietic reconstitution following transplantation, hematopoietic stem/progenitor cells (HSPC) must express genes involved in stem cell self-renewal. We investigated the expression of genes relevant for self-renewal and expansion of HSPC (operationally defined as CD34+ cells) in steady state and after transplantation. Specifically, we evaluated the expression of ninety-one genes that were analyzed by real-time PCR in CD34+ cells isolated from (i) 12 samples from umbilical cord blood (UCB); (ii) 15 samples from bone marrow healthy donors; (iii) 13 samples from bone marrow after umbilical cord blood transplant (UCBT); and (iv) 29 samples from patients after transplantation with adult hematopoietic cells. The results show that transplanted CD34+ cells from adult cells acquire an asset very different from transplanted CD34+ cells from cord blood. Multivariate machine learning analysis (MMLA) showed that four specific gene signatures can be obtained by comparing the four types of CD34+ cells. In several, but not all cases, transplanted HSPC from UCB overexpress reprogramming genes. However, these remarkable changes do not alter the commitment to hematopoietic lineage. Overall, these results reveal undisclosed aspects of transplantation biology.

JTD Keywords: Hematopoietic stem/progenitor cell, Cord blood, Stem cell transplantation


Guerrero, O., Verschure, P., (2020). Distributed adaptive control: An ideal cognitive architecture candidate for managing a robotic recycling plant Biomimetic and Biohybrid Systems 9th International Conference, Living Machines 2020 (Lecture Notes in Computer Science) , Springer International Publishing (Freiburg, Germany) 12413, 153-164

In the past decade, society has experienced notable growth in a variety of technological areas. However, the Fourth Industrial Revolution has not been embraced yet. Industry 4.0 imposes several challenges which include the necessity of new architectural models to tackle the uncertainty that open environments represent to cyber-physical systems (CPS). Waste Electrical and Electronic Equipment (WEEE) recycling plants stand for one of such open environments. Here, CPSs must work harmoniously in a changing environment, interacting with similar and not so similar CPSs, and adaptively collaborating with human workers. In this paper, we support the Distributed Adaptive Control (DAC) theory as a suitable Cognitive Architecture for managing a recycling plant. Specifically, a recursive implementation of DAC (between both single-agent and large-scale levels) is proposed to meet the expected demands of the European Project HR-Recycler. Additionally, with the aim of having a realistic benchmark for future implementations of the recursive DAC, a micro-recycling plant prototype is presented.

JTD Keywords: Cognitive architecture, Distributed Adaptive Control, Recycling plant, Navigation, Motor control, Human-Robot Interaction


Romero, D., Lázaro, J., Jané, R., Laguna, P., Bailón, R., (2020). A quaternion-based approach to estimate respiratory rate from the vectorcardiogram Computers in Cardiology (CinC) 2020 Computing in Cardiology , IEEE (Rimini, Italy) 47, 1-4

A novel ECG-derived respiration (EDR) approach is presented to efficiently estimate the respiratory rate. It combines spatial rotations and magnitude variations of the heart's electrical vector due to respiration. Orthogonal leads X, Y and Z from 10 volunteers were analyzed during a tilt table test. The largest vector magnitude (VM) within each QRS loop was assessed, and its 3D coordinates were converted into unit quaternion qb. Angular distances between these quaternions and the axes of the reference coordinate system, θ x , θ y and θ z , were then computed as EDR signals to track their relative variations caused by respiration. The respiratory rate was estimated on the spectrum of individual EDR signals obtained from the angular distances and VM time-series, but also on EDR signals obtained by principal component analysis (PCA). Relative errors (eR) to the reference respiratory signal exhibited relatively low values. The combination of EDR signals' spectrum {θ X ,θ Y, θ Z , VM} (eR=0.63±4.15%) and individual signals derived from θ X (e R =0.46±8.22%) and PCA (eR=0.36±6.58%) achieved the overall best results. The proposed method represents a computationally efficient alternative to other EDR approaches, but its robustness should be further investigated. The method could be enhanced if combined with other features tracking morphological changes induced by respiration.

JTD Keywords: Heart, Three-dimensional displays, Quaternions, Robustness, Computational efficiency, Cardiology, Principal component analysis


Mencattini, A., Di Giuseppe, D., D'Orazio, M., Rizzuto, V., Manu Pereira, M. M., Colomba Comes, M., Lopez-Martinez, M. J., Samitier, J., Martinelli, E., (2020). A microfluidic device for shape measurement in red blood cells (RBCs) IEEE International Workshop on Medical Measurement and Applications (MEMEA) , IEEE (Bari, Italy) , 1-5

Modern optical sensors coupled with time-lapse microscopy devices and dedicated software tools allow the miniaturization of laboratories for biological experiments leading to the Organ-On-Chip (OoC) framework. OoCs allow performing massive measurements on a large number of cells under the assumption of reproducibility conditions, permitting to investigate the cell dynamics in terms of motility and shape changes over time. In this work, we present the OoC platform used in a preliminary study of the Rare Haemolytic Anaemia (RHA) disease, a group of rare diseases characterized by haemolysis, which is the premature loss of red blood cells (RBCs). Preliminary results demonstrate the effectiveness of shape measurement for the diagnosis of RHA.

JTD Keywords: Anaemia diagnosis, Cell tracking, Plasticity measurement, Time-lapse microscopy


Bos, J. J., Vinck, M., Marchesi, P., Keestra, A., van Mourik-Donga, L. A., Jackson, J. C., Verschure, P., Pennartz, C. M. A., (2019). Multiplexing of self and other information in hippocampal ensembles Cell Reports 29, (12), 3859-3871.e6

In addition to coding a subject’s location in space, the hippocampus has been suggested to code social information, including the spatial position of conspecifics. “Social place cells” have been reported for tasks in which an observer mimics the behavior of a demonstrator. We examine whether rat hippocampal neurons may encode the behavior of a minirobot, but without requiring the animal to mimic it. Rather than finding social place cells, we observe that robot behavioral patterns modulate place fields coding animal position. This modulation may be confounded by correlations between robot movement and changes in the animal’s position. Although rat position indeed significantly predicts robot behavior, we find that hippocampal ensembles code additional information about robot movement patterns. Fast-spiking interneurons are particularly informative about robot position and global behavior. In conclusion, when the animal’s own behavior is conditional on external agents, the hippocampus multiplexes information about self and others.

JTD Keywords: CA1, Decoding, Information theory, Interneuron, Mutual information, Place cells, Place field, Tobot, Docial behavior, Tetrode


Vukomanovic, M., Torrents, E., (2019). High time resolution and high signal-to-noise monitoring of the bacterial growth kinetics in the presence of plasmonic nanoparticles Journal of Nanobiotechnology 17, (1), 21

Background: Emerging concepts for designing innovative drugs (i.e., novel generations of antimicrobials) frequently include nanostructures, new materials, and nanoparticles (NPs). Along with numerous advantages, NPs bring limitations, partly because they can limit the analytical techniques used for their biological and in vivo validation. From that standpoint, designing innovative drug delivery systems requires advancements in the methods used for their testing and investigations. Considering the well-known ability of resazurin-based methods for rapid detection of bacterial metabolisms with very high sensitivity, in this work we report a novel optimization for tracking bacterial growth kinetics in the presence of NPs with specific characteristics, such as specific optical properties. Results: Arginine-functionalized gold composite (HAp/Au/arginine) NPs, used as the NP model for validation of the method, possess plasmonic properties and are characterized by intensive absorption in the UV/vis region with a surface plasmon resonance maximum at 540 nm. Due to the specific optical properties, the NP absorption intensively interferes with the light absorption measured during the evaluation of bacterial growth (optical density; OD600). The results confirm substantial nonspecific interference by NPs in the signal detected during a regular turbidity study used for tracking bacterial growth. Instead, during application of a resazurin-based method (Presto Blue), when a combination of absorption and fluorescence detection is applied, a substantial increase in the signal-to-noise ratio is obtained that leads to the improvement of the accuracy of the measurements as verified in three bacterial strains tested with different growth rates (E. coli, P. aeruginosa, and S. aureus). Conclusions: Here, we described a novel procedure that enables the kinetics of bacterial growth in the presence of NPs to be followed with high time resolution, high sensitivity, and without sampling during the kinetic study. We showed the applicability of the Presto Blue method for the case of HAp/Au/arginine NPs, which can be extended to various types of metallic NPs with similar characteristics. The method is a very easy, economical, and reliable option for testing NPs designed as novel antimicrobials.

JTD Keywords: Antimicrobial nanoparticles, Arginine-functionalized gold, Bacterial growth kinetics, Plasmonic nanoparticles, Presto Blue


Roux, Anabel-Lise Lee, Quiroga, Xarxa, Walani, Nikhil, Arroyo, Marino, Roca-Cusachs, Pere, (2019). The plasma membrane as a mechanochemical transducer Philosophical Transactions of the Royal Society B: Biological Sciences 374, (1779), 20180221

Cells are constantly submitted to external mechanical stresses, which they must withstand and respond to. By forming a physical boundary between cells and their environment that is also a biochemical platform, the plasma membrane (PM) is a key interface mediating both cellular response to mechanical stimuli, and subsequent biochemical responses. Here, we review the role of the PM as a mechanosensing structure. We first analyse how the PM responds to mechanical stresses, and then discuss how this mechanical response triggers downstream biochemical responses. The molecular players involved in PM mechanochemical transduction include sensors of membrane unfolding, membrane tension, membrane curvature or membrane domain rearrangement. These sensors trigger signalling cascades fundamental both in healthy scenarios and in diseases such as cancer, which cells harness to maintain integrity, keep or restore homeostasis and adapt to their external environment.

JTD Keywords: Plasma membrane, Mechanotransduction, Membrane tension, Mechanosensor


Valenti, S., Diaz, A., Romanini, M., del Valle, L. J., Puiggalí, J., Tamarit, J. L., Macovez, R., (2019). Amorphous binary dispersions of chloramphenicol in enantiomeric pure and racemic poly-lactic acid: Morphology, molecular relaxations, and controlled drug release International Journal of Pharmaceutics 568, 118565

We characterize amorphous solid dispersions (ASDs) of the Chloramphenicol antibiotic in two biodegradable polylactic acid polymers, namely a commercial sample of enantiomeric pure PLLA and a home-synthesized PDLLA copolymer, investigating in particular the effect of polylactic acid in stabilizing the amorphous form of the drug and controlling its release (e.g. for antitumoral purposes). Broadband dielectric spectroscopy and differential scanning calorimetry are employed to study the homogeneity, glass transition temperature and relaxation dynamics of solvent-casted ASD membranes with different drug concentrations. We observe improved physical stability of the ASDs with respect to the pure drug, as well as a plasticizing effect of the antibiotic on the polymer, well described by the Gordon-Taylor equation. The release of the active pharmaceutical ingredient from the films in a simulated body fluid is studied by UV/vis spectroscopy at two different drug concentrations (5 and 20% in weight). The amount of released drug is found to be proportional to the square root of time, with proportionality constant that is almost the same in both dispersions, despite the fact that the relaxation time and thus the viscosity of the two samples differ by four orders of magnitude at body temperature. Since the drug release kinetics does not display a significant dependence on the drug content in the carrier, it may be expected to remain roughly constant during longer release times.

JTD Keywords: Amorphous drug, Controlled liberation, Dielectric spectroscopy, Molecular mobility, Plasticizer, Polymer enantiomerism


Biosca, A., Dirscherl, L., Moles, E., Imperial, S., Fernàndez-Busquets, X., (2019). An immunoPEGliposome for targeted antimalarial combination therapy at the nanoscale Pharmaceutics 11, (7), 341

Combination therapies, where two drugs acting through different mechanisms are administered simultaneously, are one of the most efficient approaches currently used to treat malaria infections. However, the different pharmacokinetic profiles often exhibited by the combined drugs tend to decrease treatment efficacy as the compounds are usually eliminated from the circulation at different rates. To circumvent this obstacle, we have engineered an immunoliposomal nanovector encapsulating hydrophilic and lipophilic compounds in its lumen and lipid bilayer, respectively. The antimalarial domiphen bromide has been encapsulated in the liposome membrane with good efficiency, although its high IC50 of ca. 1 μM for living parasites complicates its use as immunoliposomal therapy due to erythrocyte agglutination. The conjugation of antibodies against glycophorin A targeted the nanocarriers to Plasmodium-infected red blood cells and to gametocytes, the sole malaria parasite stage responsible for the transmission from the human to the mosquito vector. The antimalarials pyronaridine and atovaquone, which block the development of gametocytes, have been co-encapsulated in glycophorin A-targeted immunoliposomes. The co-immunoliposomized drugs have activities significantly higher than their free forms when tested in in vitro Plasmodium falciparum cultures: Pyronaridine and atovaquone concentrations that, when encapsulated in immunoliposomes, resulted in a 50% inhibition of parasite growth had no effect on the viability of the pathogen when used as free drugs.

JTD Keywords: Combination therapy, Immunoliposomes, Malaria, Nanomedicine, Nanotechnology, Plasmodium, Targeted drug delivery


Labay, C., Hamouda, I., Tampieri, F., Ginebra, M. P., Canal, C., (2019). Production of reactive species in alginate hydrogels for cold atmospheric plasma-based therapies Scientific Reports 9, (1), 16160

In the last years, great advances have been made in therapies based in cold atmospheric plasmas (CAP). CAP generate reactive oxygen and nitrogen species (RONS) which can be transferred to liquids. These CAP activated liquids display the same biological efficacy (i.e. on killing cancer cells) as CAP themselves, opening the door for minimally invasive therapies. However, injection of a liquid in the body results in fast diffusion due to extracellular fluids and blood flow. Therefore, the development of efficient vehicles which allow local confinement and delivery of RONS to the diseased site is a fundamental requirement. In this work, we investigate the generation of RONS (H2O2, NO2−, short-lived RONS) in alginate hydrogels by comparing two atmospheric pressure plasma jets: kINPen and a helium needle, at a range of plasma treatment conditions (time, gas flow, distance to the sample). The physic-chemical properties of the hydrogels remain unchanged by the plasma treatment, while the hydrogel shows several-fold larger capacity for generation of RONS than a typical isotonic saline solution. Part of the RONS are quickly released to a receptor media, so special attention has to be put on the design of hydrogels with in-situ crosslinking. Remarkably, the hydrogels show capacity for sustained release of the RONS. The plasma-treated hydrogels remain fully biocompatible (due the fact that the species generated by plasma are previously washed away), indicating that no cytotoxic modifications have occurred on the polymer. Moreover, the RONS generated in alginate solutions showed cytotoxic potential towards bone cancer cells. These results open the door for the use of hydrogel-based biomaterials in CAP-associated therapies.

JTD Keywords: Biomedical materials, Plasma physics


Maier, Martina, Ballester, Belén Rubio, Verschure, P., (2019). Principles of neurorehabilitation after stroke based on motor learning and brain plasticity mechanisms Frontiers in Systems Neuroscience 13, 74

What are the principles underlying effective neurorehabilitation? The aim of neurorehabilitation is to exploit interventions based on human and animal studies about learning and adaptation, as well as to show that the activation of experience-dependent neuronal plasticity augments functional recovery after stroke. Instead of teaching compensatory strategies that do not reduce impairment but allow the patient to return home as soon as possible, functional recovery might be more sustainable as it ensures a long-term reduction in impairment and an improvement in quality of life. At the same time, neurorehabilitation permits the scientific community to collect valuable data, which allows inferring about the principles of brain organization. Hence neuroscience sheds light on the mechanisms of learning new functions or relearning lost ones. However, current rehabilitation methods lack the exact operationalization of evidence gained from skill learning literature, leading to an urgent need to bridge motor learning theory and present clinical work in order to identify a set of ingredients and practical applications that could guide future interventions. This work aims to unify the neuroscientific literature relevant to the recovery process and rehabilitation practice in order to provide a synthesis of the principles that constitute an effective neurorehabilitation approach. Previous attempts to achieve this goal either focused on a subset of principles or did not link clinical application to the principles of motor learning and recovery. We identified 15 principles of motor learning based on existing literature: massed practice, spaced practice, dosage, task-specific practice, goal-oriented practice, variable practice, increasing difficulty, multisensory stimulation, rhythmic cueing, explicit feedback/knowledge of results, implicit feedback/knowledge of performance, modulate effector selection, action observation/embodied practice, motor imagery, and social interaction. We comment on trials that successfully implemented these principles and report evidence from experiments with healthy individuals as well as clinical work.

JTD Keywords: Neurorehabilitation, Motor learning, Plasticity, Stroke, Principles


Aguiar, L., Biosca, A., Lantero, E., Gut, J., Vale, N., Rosenthal, P. J., Nogueira, F., Andreu, D., Fernàndez-Busquets, X., Gomes, P., (2019). Coupling the antimalarial cell penetrating peptide TP10 to classical antimalarial drugs primaquine and chloroquine produces strongly hemolytic conjugates Molecules 24, (24), 4559

Recently, we disclosed primaquine cell penetrating peptide conjugates that were more potent than parent primaquine against liver stage Plasmodium parasites and non-toxic to hepatocytes. The same strategy was now applied to the blood-stage antimalarial chloroquine, using a wide set of peptides, including TP10, a cell penetrating peptide with intrinsic antiplasmodial activity. Chloroquine-TP10 conjugates displaying higher antiplasmodial activity than the parent TP10 peptide were identified, at the cost of an increased hemolytic activity, which was further confirmed for their primaquine analogues. Fluorescence microscopy and flow cytometry suggest that these drug-peptide conjugates strongly bind, and likely destroy, erythrocyte membranes. Taken together, the results herein reported put forward that coupling antimalarial aminoquinolines to cell penetrating peptides delivers hemolytic conjugates. Hence, despite their widely reported advantages as carriers for many different types of cargo, from small drugs to biomacromolecules, cell penetrating peptides seem unsuitable for safe intracellular delivery of antimalarial aminoquinolines due to hemolysis issues. This highlights the relevance of paying attention to hemolytic effects of cell penetrating peptide-drug conjugates.

JTD Keywords: Antimalarial, Cell penetrating peptide, Chloroquine, Erythrocyte fluorescence, Flow cytometry, Hemolysis, Microscopy, Plasmodium, Primaquine, Red blood cell


Tahirbegi, I. B., Pérez, Y., Mir, M., Samitier, J., (2019). Counterions effect on uracil-silver coordination Inorganica Chimica Acta 490, 246-253

Cyanide based silver electroplating is a low-cost reliable and well-established process for metal deposition. However, delicate handling during the process is needed because of the high toxicity of cyanide, for the persons and the environment. Uracil based silver electrodeposition got the attention of this field, because of its low cost and non-toxic nature. However, little is known about the silver complexation with uracil and the process behind the silver electroplating. In this work, we studied a hitherto unknown phenomenon on the diverse structure’s formation of silver uracil coordination complex due to the presence of different alkaline counterions. The distinct structuration of this complex clearly impacts on the efficiency and deposition yields of silver electroplating. We demonstrate the unknown key role that play hydroxide counterions in the uracil-silver coordination, and the different molecular structures created on the basis of the used counterion. The hydroxide counterion determines monomeric and polymeric complex formation with silver, which affects the solubility of the uracil silver complex and its subsequent electrodeposition. The different molecular complexes were characterized by FT-IR, UV–vis, DRUV–vis and multi-nuclear NMR spectroscopy and the silver electrodeposition by cyclic voltammetry and TOF-SIMS. This study sheds some light in the improvement of silver electroplating process

JTD Keywords: Coordination complex, Electrometallization, Electroplating, Metal complex, Silver electrodeposition, Uracil


Ballester, B. R., Maier, M., Duff, A., Cameirão, M., Bermúdez, S., Duarte, E., Cuxart, A., Rodríguez, S., San Segundo Mozo, R. M., Verschure, P., (2019). A critical time window for recovery extends beyond one-year post-stroke Journal of neurophysiology Journal of Neurophysiology , 122, (1), 350-357

The impact of rehabilitation on post-stroke motor recovery and its dependency on the patient's chronicity remain unclear. The field has widely accepted the notion of a proportional recovery rule with a "critical window for recovery" within the first 3-6 mo poststroke. This hypothesis justifies the general cessation of physical therapy at chronic stages. However, the limits of this critical window have, so far, been poorly defined. In this analysis, we address this question, and we further explore the temporal structure of motor recovery using individual patient data from a homogeneous sample of 219 individuals with mild to moderate upper-limb hemiparesis. We observed that improvement in body function and structure was possible even at late chronic stages. A bootstrapping analysis revealed a gradient of enhanced sensitivity to treatment that extended beyond 12 mo poststroke. Clinical guidelines for rehabilitation should be revised in the context of this temporal structure. NEW & NOTEWORTHY Previous studies in humans suggest that there is a 3- to 6-mo "critical window" of heightened neuroplasticity poststroke. We analyze the temporal structure of recovery in patients with hemiparesis and uncover a precise gradient of enhanced sensitivity to treatment that expands far beyond the limits of the so-called critical window. These findings highlight the need for providing therapy to patients at the chronic and late chronic stages.

JTD Keywords: Motor recovery, Neuroplasticity, Neurorehabilitation, Stroke recovery, Virtual reality


Gil, V., Del Río, J. A., (2019). Generation of 3-d collagen-based hydrogels to analyze axonal growth and behavior during nervous system development Journal of Visualized Experiments , (148), e59481

This protocol uses natural type I collagen to generate three-dimensional (3-D) hydrogel for monitoring and analyzing the axonal growth. The protocol is centered on culturing small pieces of embryonic or early postnatal rodent brains inside a 3-D hydrogel formed by the rat tail tendon-derived type I collagen with specific porosity. Tissue pieces are cultured inside the hydrogel and confronted to specific brain fragments or genetically-modified cell aggregates to produce and secrete molecules suitable for creating a gradient inside the porous matrix. The steps of this protocol are simple and reproducible but include critical steps to be considered carefully during its development. Moreover, the behavior of growing axons can be monitored and analyzed directly using a phase-contrast microscope or mono/multiphoton fluorescence microscope after fixation by immunocytochemical methods.

JTD Keywords: 3-D hydrogel cultures, Axonal growth, Cell transfection, Chemoattraction, Chemorepulsion, Embryonic nervous system, Issue 148, Neuroscience, Tissue explants


Sebastian, P., Giannotti, M. I., Gómez, E., Feliu, J. M., (2018). Surface sensitive nickel electrodeposition in deep eutectic solvent ACS Applied Energy Materials , 1, (3), 1016-1028

The first steps of nickel electrodeposition in a deep eutectic solvent (DES) are analyzed in detail. Several substrates from glassy carbon to Pt(111) were investigated pointing out the surface sensitivity of the nucleation and growth mechanism. For that, cyclic voltammetry and chronoamperometry, in combination with scanning electron microscopy (SEM), were employed. X-ray diffraction (XRD) and atomic force microscopy (AFM) were used to more deeply analyze the Ni deposition on Pt substrates. In a 0.1 M NiCl2 + DES solution (at 70 °C), the nickel deposition on glassy carbon takes place within the potential limits of the electrode in the blank solution. Although, the electrochemical window of Pt|DES is considerably shorter than on glassy carbon|DES, it was still sufficient for the nickel deposition. On the Pt electrode, the negative potential limit was enlarged while the nickel deposit grew, likely because of the lower catalytic activity of the nickel toward the reduction of the DES. At lower overpotentials, different hydrogenated Ni structures were favored, most likely because of the DES co-reduction on the Pt substrate. Nanometric metallic nickel grains of rounded shape were obtained on any substrate, as evidenced by the FE-SEM. Passivation phenomena, related to the formation of Ni oxide and Ni hydroxylated species, were observed at high applied overpotentials. At low deposited charge, on Pt(111) the AFM measurements showed the formation of rounded nanometric particles of Ni, which rearranged and formed small triangular arrays at sufficiently low applied overpotential. This particle pattern was induced by the (111) orientation and related to surface sensitivity of the nickel deposition in DES. The present work provides deep insights into the Ni electrodeposition mechanism in the selected deep eutectic solvent.

JTD Keywords: AFM, Deep eutectic solvent, Glassy carbon, Nanostructures, Nickel electrodeposition, Platinum electrode, Pt(111), SEM, Surface sensitive


Quiliano, Miguel, Pabón, Adriana, Moles, Ernest, Bonilla-Ramirez, Leonardo, Fabing, Isabelle, Fong, Kim Y., Nieto-Aco, Diego A., Wright, David W., Pizarro, Juan C., Vettorazzi, Ariane, López de Cerain, Adela, Deharo, Eric, Fernàndez-Busquets, Xavier, Garavito, Giovanny, Aldana, Ignacio, Galiano, Silvia, (2018). Structure-activity relationship of new antimalarial 1-aryl-3-susbtituted propanol derivatives: Synthesis, preliminary toxicity profiling, parasite life cycle stage studies, target exploration, and targeted delivery European Journal of Medicinal Chemistry 152, 489-514

Design, synthesis, structure-activity relationship, cytotoxicity studies, in silico drug-likeness, genotoxicity screening, and in vivo studies of new 1-aryl-3-substituted propanol derivatives led to the identification of nine compounds with promising in vitro (55, 56, 61, 64, 66, and 70–73) and in vivo (66 and 72) antimalarial profiles against Plasmodium falciparum and Plasmodium berghei. Compounds 55, 56, 61, 64, 66 and 70–73 exhibited potent antiplasmodial activity against chloroquine-resistant strain FCR-3 (IC50s < 0.28 μM), and compounds 55, 56, 64, 70, 71, and 72 showed potent biological activity in chloroquine-sensitive and multidrug-resistant strains (IC50s < 0.7 μM for 3D7, D6, FCR-3 and C235). All of these compounds share appropriate drug-likeness profiles and adequate selectivity indexes (77 < SI < 184) as well as lack genotoxicity. In vivo efficacy tests in a mouse model showed compounds 66 and 72 to be promising candidates as they exhibited significant parasitemia reductions of 96.4% and 80.4%, respectively. Additional studies such as liver stage and sporogony inhibition, target exploration of heat shock protein 90 of P. falciparum, targeted delivery by immunoliposomes, and enantiomer characterization were performed and strongly reinforce the hypothesis of 1-aryl-3-substituted propanol derivatives as promising antimalarial compounds.

JTD Keywords: Antiplasmodial, Antimalarial, Arylamino alcohol, Multi-stage activity, Hsp90, Enantiomer separation


Martí Coma-Cros, E., Biosca, A., Marques, J., Carol, L., Urbán, P., Berenguer, D., Riera, M. C., Delves, M., Sinden, R. E., Valle-Delgado, J. J., Spanos, L., Siden-Kiamos, I., Pérez, P., Paaijmans, K., Rottmann, M., Manfredi, A., Ferruti, P., Ranucci, E., Fernàndez-Busquets, X., (2018). Polyamidoamine nanoparticles for the oral administration of antimalarial drugs Pharmaceutics 10, (4), 225

Current strategies for the mass administration of antimalarial drugs demand oral formulations to target the asexual Plasmodium stages in the peripheral bloodstream, whereas recommendations for future interventions stress the importance of also targeting the transmission stages of the parasite as it passes between humans and mosquitoes. Orally administered polyamidoamine (PAA) nanoparticles conjugated to chloroquine reached the blood circulation and cured Plasmodium yoelii-infected mice, slightly improving the activity of the free drug and inducing in the animals immunity against malaria. Liquid chromatography with tandem mass spectrometry analysis of affinity chromatography-purified PAA ligands suggested a high adhesiveness of PAAs to Plasmodium falciparum proteins, which might be the mechanism responsible for the preferential binding of PAAs to Plasmodium-infected erythrocytes vs. non-infected red blood cells. The weak antimalarial activity of some PAAs was found to operate through inhibition of parasite invasion, whereas the observed polymer intake by macrophages indicated a potential of PAAs for the treatment of certain coinfections such as Plasmodium and Leishmania. When fluorescein-labeled PAAs were fed to females of the malaria mosquito vectors Anopheles atroparvus and Anopheles gambiae, persistent fluorescence was observed in the midgut and in other insect’s tissues. These results present PAAs as a versatile platform for the encapsulation of orally administered antimalarial drugs and for direct administration of antimalarials to mosquitoes, targeting mosquito stages of Plasmodium.

JTD Keywords: Anopheles, Antimalarial drugs, Malaria, Mosquitoes, Nanomedicine, Nanotechnology, Plasmodium, Polyamidoamines, Polymers, Targeted drug delivery


Guillem-Marti, J., Boix-Lemonche, G., Gugutkov, D., Ginebra, M.-P., Altankov, G., Manero, J.M., (2018). Recombinant fibronectin fragment III8-10/polylactic acid hybrid nanofibers enhance the bioactivity of titanium surface Nanomedicine 13, (8), 899-912

Aim: To develop a nanofiber (NF)-based biomimetic coating on titanium (Ti) that mimics the complex spatiotemporal organization of the extracellular matrix (ECM). Materials & methods: Recombinant cell attachment site (CAS) of fibronectin type III8-10 domain was co-electrospun with polylactic acid (PLA) and covalently bound on polished Ti discs. Osteoblast-like SaOS-2 cells were used to evaluate their complex bioactivity. Results: A significant increase of cell spreading was found on CAS/PLA hybrid NFs, followed by control pure PLA NFs and bare Ti discs. Cell proliferation showed similar trend being about twice higher on CAS/PLA NFs. The significantly increased ALP activity at day 21 indicated an enhanced differentiation of SaOS-2 cells. Conclusion: Coating of Ti implants with hybrid CAS/PLA NFs may improve significantly their osseointegration potential.

JTD Keywords: Electrospinning, Fibronectin, Hybrid nanofibers, Osseointegration, PLA, Recombinant protein


Pallarès, Irantzu, de Groot, Natalia S., Iglesias, Valentín, Sant'Anna, Ricardo, Biosca, Arnau, Fernàndez-Busquets, Xavier, Ventura, Salvador, (2018). Discovering putative prion-like proteins in Plasmodium falciparum: A computational and experimental analysis Frontiers in Microbiology 9, Article 1737

Prions are a singular subset of proteins able to switch between a soluble conformation and a self-perpetuating amyloid state. Traditionally associated with neurodegenerative diseases, increasing evidence indicates that organisms exploit prion-like mechanisms for beneficial purposes. The ability to transit between conformations is encoded in the so-called prion domains, long disordered regions usually enriched in glutamine/asparagines residues. Interestingly, Plasmodium falciparum, the parasite that causes the most virulent form of malaria, is exceptionally rich in proteins bearing long Q/N-rich sequence stretches, accounting for roughly 30% of the proteome. This biased composition suggests that these protein regions might correspond to prion-like domains (PrLDs) and potentially form amyloid assemblies. To investigate this possibility, we performed a stringent computational survey for Q/N-rich PrLDs on P. falciparum. Our data indicate that ~10% of P. falciparum protein sequences have prionic signatures, and that this subproteome is enriched in regulatory proteins, such as transcription factors and RNA-binding proteins. Furthermore, we experimentally demonstrate for several of the identified PrLDs that, despite their disordered nature, they contain inner short sequences able to spontaneously self-assemble into amyloid-like structures. Although the ability of these sequences to nucleate the conformational conversion of the respective full-length proteins should still be demonstrated, our analysis suggests that, as previously described for other organisms, prion-like proteins might also play a functional role in P. falciparum.

JTD Keywords: Plasmodium, Protein aggregation, Amyloid, Prion, Q-N-rich sequences, Protein Disorder


Farré, Ramon, Otero, Jordi, Almendros, Isaac, Navajas, Daniel, (2018). Bioengineered lungs: A challenge and an opportunity Archivos de Bronconeumología 54, (1), 31-38

Lung biofabrication is a new tissue engineering and regenerative development aimed at providing organs for potential use in transplantation. Lung biofabrication is based on seeding cells into an acellular organ scaffold and on culturing them in an especial purpose bioreactor. The acellular lung scaffold is obtained by decellularizing a non-transplantable donor lung by means of conventional procedures based on application of physical, enzymatic and detergent agents. To avoid immune recipient's rejection of the transplanted bioengineered lung, autologous bone marrow/adipose tissue-derived mesenchymal stem cells, lung progenitor cells or induced pluripotent stem cells are used for biofabricating the bioengineered lung. The bioreactor applies circulatory perfusion and mechanical ventilation with physiological parameters to the lung during biofabrication. These physical stimuli to the organ are translated into the stem cell local microenvironment - e.g. shear stress and cyclic stretch - so that cells sense the physiological conditions in normally functioning mature lungs. After seminal proof of concept in a rodent model was published in 2010, the hypothesis that lungs can be biofabricated is accepted and intense research efforts are being devoted to the topic. The current experimental evidence obtained so far in animal tests and in ex vivo human bioengineered lungs suggests that the date of first clinical tests, although not immediate, is coming. Lung bioengineering is a disrupting concept that poses a challenge for improving our basic science knowledge and is also an opportunity for facilitating lung transplantation in future clinical translation.

JTD Keywords: Tissue engineering, Regenerative medicine, Lung transplantation, Lung repair, Lung regeneration


Martí Coma-Cros, Elisabet, Biosca, Arnau, Lantero, Elena, Manca, Maria, Caddeo, Carla, Gutiérrez, Lucía, Ramírez, Miriam, Borgheti-Cardoso, Livia, Manconi, Maria, Fernàndez-Busquets, Xavier, (2018). Antimalarial activity of orally administered curcumin incorporated in Eudragit®-containing liposomes International Journal of Molecular Sciences 19, (5), 1361

Curcumin is an antimalarial compound easy to obtain and inexpensive, having shown little toxicity across a diverse population. However, the clinical use of this interesting polyphenol has been hampered by its poor oral absorption, extremely low aqueous solubility and rapid metabolism. In this study, we have used the anionic copolymer Eudragit® S100 to assemble liposomes incorporating curcumin and containing either hyaluronan (Eudragit-hyaluronan liposomes) or the water-soluble dextrin Nutriose® FM06 (Eudragit-nutriosomes). Upon oral administration of the rehydrated freeze-dried nanosystems administered at 25/75 mg curcumin·kg−1·day−1, only Eudragit-nutriosomes improved the in vivo antimalarial activity of curcumin in a dose-dependent manner, by enhancing the survival of all Plasmodium yoelii-infected mice up to 11/11 days, as compared to 6/7 days upon administration of an equal dose of the free compound. On the other hand, animals treated with curcumin incorporated in Eudragit-hyaluronan liposomes did not live longer than the controls, a result consistent with the lower stability of this formulation after reconstitution. Polymer-lipid nanovesicles hold promise for their development into systems for the oral delivery of curcumin-based antimalarial therapies.

JTD Keywords: Malaria, Curcumin, Nanomedicine, Oral administration, Lipid nanovesicles, Eudragit, Nutriose, Hyaluronan, Plasmodium yoelii


Khurana, Kanupriya, Müller, Frank, Jacobs, Karin, Faidt, Thomas, Neurohr, Jens-Uwe, Grandthyll, Samuel, Mücklich, Frank, Canal, Cristina, Pau Ginebra, Maria, (2018). Plasma polymerized bioceramics for drug delivery: Do surface changes alter biological behaviour? European Polymer Journal 107, 25-33

One of the treatments for recurrent or complicated osteomyelitis is by local antibiotherapy mediated by suitable bone grafts. β–Tricalcium Phosphate (β–TCP) bioceramic is a resorbable bone graft. Its microporosity allows for incorporation of drugs, but a too fast release is often obtained. Complex strategies have been explored to obtain controlled drug release. In this work, plasma polymerization of a biocompatible polymer was investigated on β-TCP. Polyethyleneglycol (PEG)-like polymer coatings of different thickness were deposited on microporous β-TCP loaded with antibiotics. A highly hydrophobic surface was obtained despite the hydrophilicity of the PEG-like layer produced, which was associated to the roughness of the β-TCP substrate. The bioceramics nevertheless retained their suitable biological behavior with regard to human osteoblast cells. The microbiological activity of the antibiotics was preserved, and the coatings reduced the total amount of drug released as a function of the increasing plasma treatment time.

JTD Keywords: Plasma polymerization, β–Tricalcium phosphate, PEG-like polymer, Antibiotics, Drug release, Biocompatibility


Borgheti-Cardoso, L.N., Fernàndez-Busquets, X., (2018). Turning Plasmodium survival strategies against itself Future Medicinal Chemistry 10, (19), 2245-2248

Miquel, Joan, Santana, F., Palau, E., Vinagre, M., Langohr, K., Casals, A., Torrens, C., (2018). Retaining or excising the supraspinatus tendon in complex proximal humeral fractures treated with reverse prosthesis: a biomechanical analysis in two different designs Archives of Orthopaedic and Trauma Surgery 138, (11), 1533-1539

We aimed to biomechanically evaluate the effect of the supraspinatus tendon on tuberosity stability using two different reverse shoulder arthroplasty (RSA) models for complex proximal humeral fractures (PHFs).

JTD Keywords: Tuberosity reconstruction, Reverse shoulder arthroplasty, Supraspinatus, Cadaveric study, Rotator cuff excision, Complex proximal humeral fractures


Vouloutsi, Vasiliki, Halloy, José, Mura, Anna, Mangan, Michael, Lepora, Nathan, Prescott, T. J., Verschure, P., (2018). Biomimetic and Biohybrid Systems 7th International Conference, Living Machines 2018, Paris, France, July 17–20, 2018, Proceedings , Springer International Publishing (Lausanne, Switzerland) 10928, 1-551

This book constitutes the proceedings of the 7th International Conference on Biomimetic and Biohybrid Systems, Living Machines 2018, held in Paris, France, in July 2018. The 40 full and 18 short papers presented in this volume were carefully reviewed and selected from 60 submissions. The theme of the conference targeted at the intersection of research on novel life-like technologies inspired by the scientific investigation of biological systems, biomimetics, and research that seeks to interface biological and artificial systems to create biohybrid systems.

JTD Keywords: Artificial neural network, Bio-actuators, Bio-robotics, Biohybrid systems, Biomimetics, Bipedal robots, Earthoworm-like robots, Robotics, Decision-making, Tactile sensing, Soft robots, Locomotion, Insects, Sensors, Actuators, Robots, Artificial intelligence, Neural networks, Motion planning, Learning algorithms


Barbeck, Mike, Serra, Tiziano, Booms, Patrick, Stojanovic, Sanja, Najman, Stevo, Engel, Elisabeth, Sader, Robert, Kirkpatrick, Charles James, Navarro, Melba, Ghanaati, Shahram, (2017). Analysis of the in vitro degradation and the in vivo tissue response to bi-layered 3D-printed scaffolds combining PLA and biphasic PLA/bioglass components – Guidance of the inflammatory response as basis for osteochondral regeneration Bioactive Materials , 2, (4), 208-223

Abstract The aim of the present study was the in vitro and in vivo analysis of a bi-layered 3D-printed scaffold combining a PLA layer and a biphasic PLA/bioglass G5 layer for regeneration of osteochondral defects in vivo Focus of the in vitro analysis was on the (molecular) weight loss and the morphological and mechanical variations after immersion in SBF. The in vivo study focused on analysis of the tissue reactions and differences in the implant bed vascularization using an established subcutaneous implantation model in CD-1 mice and established histological and histomorphometrical methods. Both scaffold parts kept their structural integrity, while changes in morphology were observed, especially for the PLA/G5 scaffold. Mechanical properties decreased with progressive degradation, while the PLA/G5 scaffolds presented higher compressive modulus than PLA scaffolds. The tissue reaction to PLA included low numbers of BMGCs and minimal vascularization of its implant beds, while the addition of G5 lead to higher numbers of BMGCs and a higher implant bed vascularization. Analysis revealed that the use of a bi-layered scaffold shows the ability to observe distinct in vivo response despite the physical proximity of PLA and PLA/G5 layers. Altogether, the results showed that the addition of G5 enables to reduce scaffold weight loss and to increase mechanical strength. Furthermore, the addition of G5 lead to a higher vascularization of the implant bed required as basis for bone tissue regeneration mediated by higher numbers of BMGCs, while within the PLA parts a significantly lower vascularization was found optimally for chondral regeneration. Thus, this data show that the analyzed bi-layered scaffold may serve as an ideal basis for the regeneration of osteochondral tissue defects. Additionally, the results show that it might be able to reduce the number of experimental animals required as it may be possible to analyze the tissue response to more than one implant in one experimental animal.

JTD Keywords: Bioactive glass, Polylactic acid (PLA), Bi-layer scaffold, Multinucleated giant cells, Bone substitute, Vascularization, Calcium phosphate glass


Moles, E., Galiano, S., Gomes, A., Quiliano, M., Teixeira, C., Aldana, I., Gomes, P., Fernàndez-Busquets, X., (2017). ImmunoPEGliposomes for the targeted delivery of novel lipophilic drugs to red blood cells in a falciparum malaria murine model Biomaterials 145, 178-191

Most drugs currently entering the clinical pipeline for severe malaria therapeutics are of lipophilic nature, with a relatively poor solubility in plasma and large biodistribution volumes. Low amounts of these compounds do consequently accumulate in circulating Plasmodium-infected red blood cells, exhibiting limited antiparasitic activity. These drawbacks can in principle be satisfactorily dealt with by stably encapsulating drugs in targeted nanocarriers. Here this approach has been adapted for its use in immunocompetent mice infected by the Plasmodium yoelii 17XL lethal strain, selected as a model for human blood infections by Plasmodium falciparum. Using immunoliposomes targeted against a surface protein characteristic of the murine erythroid lineage, the protocol has been applied to two novel antimalarial lipophilic drug candidates, an aminoquinoline and an aminoalcohol. Large encapsulation yields of >90% were obtained using a citrate-buffered pH gradient method and the resulting immunoliposomes reached in vivo erythrocyte targeting and retention efficacies of >80%. In P. yoelii-infected mice, the immunoliposomized aminoquinoline succeeded in decreasing blood parasitemia from severe to uncomplicated malaria parasite densities (i.e. from ≥25% to ca. 5%), whereas the same amount of drug encapsulated in non-targeted liposomes had no significant effect on parasite growth. Pharmacokinetic analysis indicated that this good performance was obtained with a rapid clearance of immunoliposomes from the circulation (blood half-life of ca. 2 h), suggesting a potential for improvement of the proposed model.

JTD Keywords: Immunoliposomes, Malaria, Nanomedicine, Plasmodium falciparum, Plasmodium yoelii 17XL, Targeted drug delivery


Marques, J., Valle-Delgado, J. J., Urbán, P., Baró, E., Prohens, R., Mayor, A., Cisteró, P., Delves, M., Sinden, R. E., Grandfils, C., de Paz, J. L., García-Salcedo, J. A., Fernàndez-Busquets, X., (2017). Adaptation of targeted nanocarriers to changing requirements in antimalarial drug delivery Nanomedicine: Nanotechnology, Biology, and Medicine 13, (2), 515-525

The adaptation of existing antimalarial nanocarriers to new Plasmodium stages, drugs, targeting molecules, or encapsulating structures is a strategy that can provide new nanotechnology-based, cost-efficient therapies against malaria. We have explored the modification of different liposome prototypes that had been developed in our group for the targeted delivery of antimalarial drugs to Plasmodium-infected red blood cells (pRBCs). These new models include: (i) immunoliposome-mediated release of new lipid-based antimalarials; (ii) liposomes targeted to pRBCs with covalently linked heparin to reduce anticoagulation risks; (iii) adaptation of heparin to pRBC targeting of chitosan nanoparticles; (iv) use of heparin for the targeting of Plasmodium stages in the mosquito vector; and (v) use of the non-anticoagulant glycosaminoglycan chondroitin 4-sulfate as a heparin surrogate for pRBC targeting. The results presented indicate that the tuning of existing nanovessels to new malaria-related targets is a valid low-cost alternative to the de novo development of targeted nanosystems.

JTD Keywords: Glycosaminoglycans, Malaria, Nanomedicine, Plasmodium, Targeted drug delivery


Canal, C., Fontelo, R., Hamouda, I., Guillem-Marti, J., Cvelbar, U., Ginebra, M. P., (2017). Plasma-induced selectivity in bone cancer cells death Free Radical Biology and Medicine , 110, 72-80

Background: Current therapies for bone cancers - either primary or metastatic – are difficult to implement and unfortunately not completely effective. An alternative therapy could be found in cold plasmas generated at atmospheric pressure which have already demonstrated selective anti-tumor action in a number of carcinomas and in more relatively rare brain tumors. However, its effects on bone cancer are still unknown. Methods: Herein, we employed an atmospheric pressure plasma jet (APPJ) to validate its selectivity towards osteosarcoma cell line vs. osteoblasts & human mesenchymal stem cells. Results: Cytotoxicity following direct interaction of APPJ with cells is comparable to indirect interaction when only liquid medium is treated and subsequently added to the cells, especially on the long-term (72 h of cell culture). Moreover, following contact of the APPJ treated medium with cells, delayed effects are observed which lead to 100% bone cancer cell death through apoptosis (decreased cell viability with incubation time in contact with APPJ treated medium from 24 h to 72 h), while healthy cells remain fully viable and unaffected by the treatment. Conclusions: The high efficiency of the indirect treatment indicates that an important role is played by the reactive oxygen species (ROS) and reactive nitrogen species (RNS) in the gaseous plasma stage and then transmitted to the liquid phase, which overall lead to lethal and selective action towards osteosarcoma cells. These findings open new pathways for treatment of metastatic bone disease with a minimally invasive approach.

JTD Keywords: Atmospheric pressure plasma jet, Bone cancer, hMSC, HOb, Liquids, Osteoblasts, Osteosarcoma, SaOS-2


Schieber, R., Lasserre, F., Hans, M., Fernández-Yagüe, M., Díaz-Ricart, M., Escolar, G., Ginebra, M. P., Mücklich, F., Pegueroles, M., (2017). Direct laser interference patterning of CoCr alloy surfaces to control endothelial cell and platelet response for cardiovascular applications Advanced Healthcare Materials 6, (19), 1700327

The main drawbacks of cardiovascular bare-metal stents (BMS) are in-stent restenosis and stent thrombosis as a result of an incomplete endothelialization after stent implantation. Nano- and microscale modification of implant surfaces is a strategy to recover the functionality of the artery by stimulating and guiding molecular and biological processes at the implant/tissue interface. In this study, cobalt-chromium (CoCr) alloy surfaces are modified via direct laser interference patterning (DLIP) in order to create linear patterning onto CoCr surfaces with different periodicities (≈3, 10, 20, and 32 μm) and depths (≈20 and 800 nm). Changes in surface topography, chemistry, and wettability are thoroughly characterized before and after modification. Human umbilical vein endothelial cells' adhesion and spreading are similar for all patterned and plain CoCr surfaces. Moreover, high-depth series induce cell elongation, alignment, and migration along the patterned lines. Platelet adhesion and aggregation decrease in all patterned surfaces compared to CoCr control, which is associated with changes in wettability and oxide layer characteristics. Cellular studies provide evidence of the potential of DLIP topographies to foster endothelialization without enhancement of platelet adhesion, which will be of high importance when designing new BMS in the future.

JTD Keywords: CoCr, Direct laser interference patterning, Endothelial cells, Linear surface pattern, Platelets


Aláez-Versón, C. R., Lantero, E., Fernàndez-Busquets, X., (2017). Heparin: New life for an old drug Nanomedicine 12, (14), 1727-1744

Heparin is one of the oldest drugs, which nevertheless remains in widespread clinical use as an inhibitor of blood coagulation. The history of its identification a century ago unfolded amid one of the most fascinating scientific controversies turning around the distribution of credit for its discovery. The composition, purification and structure-function relationship of this naturally occurring glycosaminoglycan regarding its classical role as anticoagulant will be dealt with before proceeding to discuss its therapeutic potential in, among other, inflammatory and infectious disease, cancer treatment, cystic fibrosis and Alzheimer's disease. The first bibliographic reference hit using the words 'nanomedicine' and 'heparin' is as recent as 2008. Since then, nanomedical applications of heparin have experienced an exponential growth that will be discussed in detail, with particular emphasis on its antimalarial activity. Some of the most intriguing potential applications of heparin nanomedicines will be exposed, such as those contemplating the delivery of drugs to the mosquito stages of malaria parasites.

JTD Keywords: Anopheles, Antimalarial drugs, Heparin, Malaria, Mosquitoes, Nanomedicine, Nanotechnology, Plasmodium, Targeted drug delivery


Moles, E., Marcos, J., Imperial, S., Pozo, O. J., Fernàndez-Busquets, X., (2017). 2-picolylamine derivatization for high sensitivity detection of abscisic acid in apicomplexan blood-infecting parasites Talanta 168, 130-135

We have developed a new liquid chromatography-electrospray ionization tandem mass spectrometry methodology based on 2-picolylamine derivatization and positive ion mode detection for abscisic acid (ABA) identification. The selected reaction leads to the formation of an amide derivative which contains a highly active pyridyl group. The enhanced ionization allows for a 700-fold increase over commonly monitored unmodified ABA, which in turn leads to excellent limits of detection and quantification values of 0.03 and 0.15 ng mL-1, respectively. This method has been validated in the highly complex matrix of a red blood cell extract. In spite of the high sensitivity achieved, ABA could not be detected in Plasmodium falciparum-infected red blood cells, suggesting that, if present, it will be found either in ultratrace amounts or as brief bursts at defined time points within the intraerythrocytic cycle and/or in the form of a biosynthetic analogue.

JTD Keywords: Abscisic acid, Apicomplexa, Liquid chromatography-electrospray ionization tandem mass spectrometry, Malaria, Picolylamine, Plasmodium falciparum


Punet, X., Levato, R., Bataille, I., Letourneur, D., Engel, E., Mateos-Timoneda, M. A., (2017). Polylactic acid organogel as versatile scaffolding technique Polymer 113, 81-91

Tissue engineering requires scaffolding techniques based on non-toxic processes that permits the fabrication of constructs with tailored properties. Here, a two-step methodology based on the gelation and precipitation of the poly(lactic) acid/ethyl lactate organogel system is presented. With this technique nanofibrous matrices that resemble natural extracellular matrix can be easily obtained, while allowing control over the mechanical properties of the device. Gelation temperature and the dynamics of the gelation of the organogel system are characterized, and the final mechanical and viscoelastic properties, as well as porosity, as function of the initial polymer concentration are described. We show that gelation temperature of the system is concentration independent and below 44.5 °C, which permits gelation at room temperature. Furthermore, mechanical properties are found in the range of the soft organic tissues, and the obtained micro-network architecture gives place to a flexible structure. Such structure presents tuneable elastic modulus and viscoelastic properties as function of nanofibers density. Moreover, centimetre-long tubular scaffolds with the diameter of medium-caliber blood vessels were produced. The fibrous nano-architecture mimics the native extracellular matrix fibres diameter and morphology was proven to be suitable to support endothelialization of the lumen of the tube. Thus, this strategy, based on biocompatible green compound might be promising for the fabrication of large 3D scaffolds for tissue engineering applications.

JTD Keywords: Gel, Gelation, Nanofibrous, Organogel, PLA, Poly(lactic) acid, Scaffold


Castellanos, M. I., Guillem-Marti, J., Mas-Moruno, C., Díaz-Ricart, M., Escolar, G., Ginebra, M. P., Gil, F. J., Pegueroles, M., Manero, J. M., (2017). Cell adhesive peptides functionalized on CoCr alloy stimulate endothelialization and prevent thrombogenesis and restenosis Journal of Biomedical Materials Research - Part A , 105, (4), 973-983

Immobilization of bioactive peptide sequences on CoCr surfaces is an effective route to improve endothelialization, which is of great interest for cardiovascular stents. In this work, we explored the effect of physical and covalent immoblization of RGDS, YIGSR and their equimolar combination peptides on endothelial cells (EC) and smooth muscle cell (SMC) adhesion and on thrombogenicity. We extensively investigated using RT-qPCR, the expression by ECs cultured on functionalised CoCr surfaces of different genes. Genes relevant for adhesion (ICAM-1 and VCAM-1), vascularization (VEGFA, VEGFR-1 and VEGFR-2) and anti-thrombogenicity (tPA and eNOS) were over-expressed in the ECs grown to covalently functionalized CoCr surfaces compared to physisorbed and control surfaces. Pro-thrombogenic genes expression (PAI-1 and vWF) decreased over time. Cell co-cultures of ECs/SMCs found that functionalization increased the amount of adhered ECs onto modified surfaces compared to plain CoCr, independently of the used peptide and the strategy of immobilization. SMCs adhered less compared to ECs in all surfaces. All studied peptides showed a lower platelet cell adhesion compared to TCPS. Covalent functionalization of CoCr surfaces with an equimolar combination of RGDS and YIGSR represented prevailing strategy to enhance the early stages of ECs adhesion and proliferation, while preventing SMCs and platelet adhesion.

JTD Keywords: Cell coculture, CoCr alloy, Functionalization, Gene expression, Platelet adhesion


Ballester, Rubio Belén, Nirme, Jens, Camacho, Irene, Duarte, Esther, Rodríguez, Susana, Cuxart, Ampar, Duff, Armin, Verschure, F. M. J. Paul, (2017). Domiciliary VR-based therapy for functional recovery and cortical reorganization: Randomized controlled trial in participants at the chronic stage post stroke JMIR Serious Games , 5, (3), e15

Background: Most stroke survivors continue to experience motor impairments even after hospital discharge. Virtual reality-based techniques have shown potential for rehabilitative training of these motor impairments. Here we assess the impact of at-home VR-based motor training on functional motor recovery, corticospinal excitability and cortical reorganization. Objective: The aim of this study was to identify the effects of home-based VR-based motor rehabilitation on (1) cortical reorganization, (2) corticospinal tract, and (3) functional recovery after stroke in comparison to home-based occupational therapy. Methods: We conducted a parallel-group, controlled trial to compare the effectiveness of domiciliary VR-based therapy with occupational therapy in inducing motor recovery of the upper extremities. A total of 35 participants with chronic stroke underwent 3 weeks of home-based treatment. A group of subjects was trained using a VR-based system for motor rehabilitation, while the control group followed a conventional therapy. Motor function was evaluated at baseline, after the intervention, and at 12-weeks follow-up. In a subgroup of subjects, we used Navigated Brain Stimulation (NBS) procedures to measure the effect of the interventions on corticospinal excitability and cortical reorganization. Results: Results from the system?s recordings and clinical evaluation showed significantly greater functional recovery for the experimental group when compared with the control group (1.53, SD 2.4 in Chedoke Arm and Hand Activity Inventory). However, functional improvements did not reach clinical significance. After the therapy, physiological measures obtained from a subgroup of subjects revealed an increased corticospinal excitability for distal muscles driven by the pathological hemisphere, that is, abductor pollicis brevis. We also observed a displacement of the centroid of the cortical map for each tested muscle in the damaged hemisphere, which strongly correlated with improvements in clinical scales. Conclusions: These findings suggest that, in chronic stages, remote delivery of customized VR-based motor training promotes functional gains that are accompanied by neuroplastic changes. Trial Registration: International Standard Randomized Controlled Trial Number NCT02699398 (Archived by ClinicalTrials.gov at https://clinicaltrials.gov/ct2/show/NCT02699398?term=NCT02699398&rank=1)

JTD Keywords: Stroke, Movement disorder, Recovery of function, neuroplasticity, Transcranial magnetic stimulation, Physical therapy, Hemiparesis, Computer applications software


Torras, Núria, Agusil, Juan Pablo, Vázquez, Patricia, Duch, Marta, Hernández-Pinto, Alberto M., Samitier, Josep, de la Rosa, Enrique J., Esteve, Jaume, Suárez, Teresa, Pérez-García, Lluïsa, Plaza, José A., (2016). Suspended planar-array chips for molecular multiplexing at the microscale Advanced Materials 28, (7), 1449–1454

A novel suspended planar-array chips technology is described, which effectively allows molecular multiplexing using a single suspended chip to analyze extraordinarily small volumes. The suspended chips are fabricated by combining silicon-based technology and polymer-pen lithography, obtaining increased molecular pattern flexibility, and improving miniaturization and parallel production. The chip miniaturization is so dramatic that it permits the intracellular analysis of living cells.

JTD Keywords: Chip-in-a-cell, Suspended-arrays, Planar-arrays, Silicon chips, Single-cell analysis


Giannotti, M. I., Abasolo, Ibane, Oliva, Mireia, Andrade, Fernanda, García-Aranda, Natalia, Melgarejo, Marta, Pulido, Daniel, Corchero, José Luis, Fernández, Yolanda, Villaverde, Antonio, Royo, Miriam, Garcia-Parajo, Maria F., Sanz, Fausto, Schwartz Jr, Simó, (2016). Highly versatile polyelectrolyte complexes for improving the enzyme replacement therapy of lysosomal storage disorders ACS Applied Materials & Interfaces 8, (39), 25741–25752

Lysosomal storage disorders are currently treated by enzyme replacement therapy (ERT) through the direct administration of the unprotected recombinant protein to the patients. Herein we present an ionically cross-linked polyelectrolyte complex (PEC) composed of trimethyl chitosan (TMC) and α-galactosidase A (GLA), the defective enzyme in Fabry disease, with the capability of directly targeting endothelial cells by incorporating peptide ligands containing the RGD sequence. We assessed the physicochemical properties, cytotoxicity, and hemocompatibility of RGD-targeted and untargeted PECs, the uptake by endothelial cells and the intracellular activity of PECs in cell culture models of Fabry disease. Moreover, we also explored the effect of different freeze-drying procedures in the overall activity of the PECs. Our results indicate that the use of integrin-binding RGD moiety within the PEC increases their uptake and the efficacy of the GLA enzyme, while the freeze-drying allows the activity of the therapeutic protein to remain intact. Overall, these results highlight the potential of TMC-based PECs as a highly versatile and feasible drug delivery system for improving the ERT of lysosomal storage disorders.

JTD Keywords: Enzyme replacement therapy, Fabry disease, Lysosomal delivery, Nanomedicine, Polyelectrolyte complexes, Trimethyl chitosan, α-galactosidase A


Fernàndez-Busquets, X., (2016). Novel strategies for Plasmodium-targeted drug delivery Expert Opinion on Drug Delivery , 13, (7), 919-922

Noguera-Ortega, E., Rabanal, R. M., Secanella-Fandos, S., Torrents, E., Luquin, M., Julián, E., (2016). Gamma-irradiated mycobacteria enhance survival in bladder tumor bearing mice although less efficaciously than live mycobacteria Journal of Urology , 195, (1), 198-205

Purpose γ Irradiated Mycobacterium bovis bacillus Calmette-Guérin has shown in vitro and ex vivo antitumor activity. However, to our knowledge the potential antitumor capacity has not been demonstrated in vivo. We studied the in vivo potential of γ irradiated bacillus Calmette-Guérin and γ irradiated M. brumae, a saprophytic mycobacterium that was recently described as an immunotherapeutic agent. Materials and Methods The antitumor capacity of γ irradiated M. brumae was first investigated by analyzing the in vitro inhibition of bladder tumor cell proliferation and the ex vivo cytotoxic effect of M. brumae activated peripheral blood cells. The effect of γ irradiated M. brumae or bacillus Calmette-Guérin intravesical treatment was then compared to treatment with live mycobacteria in the orthotopic murine model of bladder cancer. Results Nonviable M. brumae showed a capacity to inhibit in vitro bladder cancer cell lines similar to that of live mycobacteria. However, its capacity to induce cytokine production was decreased compared to that of live M. brumae. γ Irradiated M. brumae could activate immune cells to inhibit tumor cell growth, although to a lesser extent than live mycobacteria. Finally, intravesical treatment with γ irradiated M. brumae or bacillus Calmette-Guérin significantly increased survival with respect to that of nontreated tumor bearing mice. Both γ irradiated mycobacteria showed lower survival rates than those of live mycobacteria but the minor efficacy of γ irradiated vs live mycobacteria was only significant for bacillus Calmette-Guérin. Conclusions Our results show that although γ irradiated mycobacteria is less efficacious than live mycobacteria, it induces an antitumor effect in vivo, avoiding the possibility of further mycobacterial infections.

JTD Keywords: BCG vaccine, Gamma rays, Immunotherapy, Mycobacterium, Urinary bladder neoplasms


Tassinari, E., Aznar, S., Urcola, I., Prieto, A., Hüttener, M., Juárez, A., (2016). The incC sequence is required for R27 plasmid stability Frontiers in Microbiology 7, (6), Article 629

IncHI plasmids account for multiple antimicrobial resistance in Salmonella and other enterobacterial genera. These plasmids are generally very stable in their bacterial hosts. R27 is the archetype of IncHI1 plasmids. A high percentage of the R27-encoded open reading frames (ORFs) (66.7%) do not show similarity to any known ORFs. We performed a deletion analysis of all non-essential R27 DNA sequences to search for hitherto non-identified plasmid functions that might be required for plasmid stability. We report the identification of a short DNA sequence (incC) that is essential for R27 stability. That region contains several repeats (incC repeats), belongs to one of the three-plasmid replicons (R27 FIA-like) and is targeted by the R27 E protein. Deletion of the incC sequence drastically reduces R27 stability both in Escherichia coli and in Salmonella, the effect being more pronounced in this latter species. Interfering with incC-E protein interaction must lead to a reduced IncHI1 plasmid stability, and may represent a new approach to combat antimicrobial resistance.

JTD Keywords: Antimicrobial resistance, E protein, IncC, IncHI1 plasmids, Plasmid R27, Plasmid stability


Valle-Delgado, J. J., Fernàndez-Busquets, X., (2016). Rapid diagnostic tests for malaria: Past, present and future Future Microbiology , 11, (11), 1379-1382

Forget, J., Awaja, F., Gugutkov, D., Gustavsson, J., Gallego Ferrer, G., Coelho-Sampaio, T., Hochman-Mendez, C., Salmeron-Sánchez, M., Altankov, G., (2016). Differentiation of human mesenchymal stem cells toward quality cartilage using fibrinogen-based nanofibers Macromolecular Bioscience 16, (9), 1348-1359

Mimicking the complex intricacies of the extra cellular matrix including 3D configurations and aligned fibrous structures were traditionally perused for producing cartilage tissue from stem cells. This study shows that human adipose derived mesenchymal stem cells (hADMSCs) establishes significant chondrogenic differentiation and may generate quality cartilage when cultured on 2D and randomly oriented fibrinogen/poly-lactic acid nanofibers compared to 3D sandwich-like environments. The adhering cells show well-developed focal adhesion complexes and actin cytoskeleton arrangements confirming the proper cellular interaction with either random or aligned nanofibers. However, quantitative reverse transcription-polymerase chain reaction analysis for Collagen 2 and Collagen 10 genes expression confirms favorable chondrogenic response of hADMSCs on random nanofibers and shows substantially higher efficacy of their differentiation in 2D configuration versus 3D constructs. These findings introduce a new direction for cartilage tissue engineering through providing a simple platform for the routine generation of transplantable stem cells derived articular cartilage replacement that might improve joint function.

JTD Keywords: Cartilage, Chondrogenic response, Collagen, FBG/PLA nanofibers, Mesenchymal stem cells


Silva, N., Muñoz, C., Diaz-Marcos, J., Samitier, J., Yutronic, N., Kogan, M. J., Jara, P., (2016). In situ visualization of the local photothermal effect produced on α-cyclodextrin inclusion compound associated with gold nanoparticles Nanoscale Research Letters 11, 180

Evidence of guest migration in α-cyclodextrin-octylamine (α-CD-OA) inclusion compound (IC) generated via plasmonic heating of gold nanoparticles (AuNPs) has been studied. In this report, we demonstrate local effects generated by laser-mediated irradiation of a sample of AuNPs covered with inclusion compounds on surface-derivatized glass under liquid conditions by atomic force microscopy (AFM). Functionalized AuNPs on the glass and covered by the ICs were monitored by recording images by AFM during 5 h of irradiation, and images showed that after irradiation, a drastic decrease in the height of the AuNPs occurred. The absorption spectrum of the irradiated sample showed a hypsochromic shift from 542 to 536 nm, evidence suggesting that much of the population of nanoparticles lost all of the parts of the overlay of ICs due to the plasmonic heat generated by the irradiation. Mass spectrometry matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) performed on a sample containing a collection of drops obtained from the surface of the functionalized glass provided evidence that the irradiation lead to disintegration of the ICs and therefore exit of the octylamine molecule (the guest) from the cyclodextrin cavity (the matrix).

JTD Keywords: Cyclodextrin inclusion compound, Gold nanoparticles, Guest migration, Plasmonic heating


Carrera, I., Gelber, P. E., Chary, G., González-Ballester, M. A., Monllau, J. C., Noailly, J., (2016). Fixation of a split fracture of the lateral tibial plateau with a locking screw plate instead of cannulated screws would allow early weight bearing: a computational exploration International Orthopaedics , 40, (10), 2163-2169

Purpose: To assess, with finite element (FE) calculations, whether immediate weight bearing would be possible after surgical stabilization either with cannulated screws or with a locking plate in a split fracture of the lateral tibial plateau (LTP). Methods: A split fracture of the LTP was recreated in a FE model of a human tibia. A three-dimensional FE model geometry of a human femur-tibia system was obtained from the VAKHUM project database, and was built from CT images from a subject with normal bone morphologies and normal alignment. The mesh of the tibia was reconverted into a geometry of NURBS surfaces. A split fracture of the lateral tibial plateau was reproduced by using geometrical data from patient radiographs. A locking screw plate (LP) and a cannulated screw (CS) systems were modelled to virtually reduce the fracture and 80 kg static body-weight was simulated. Results: While the simulated body-weight led to clinically acceptable interfragmentary motion, possible traumatic bone shear stresses were predicted nearby the cannulated screws. With a maximum estimation of about 1.7 MPa maximum bone shear stresses, the Polyax system might ensure more reasonable safety margins. Conclusions: Split fractures of the LTP fixed either with locking screw plate or cannulated screws showed no clinically relevant IFM in a FE model. The locking screw plate showed higher mechanical stability than cannulated screw fixation. The locking screw plate might also allow full or at least partial weight bearing under static posture at time zero.

JTD Keywords: Bone fixation, Finite element, Fracture fixation, Interfragmentary motion, Tibial plateau fractures, Weight bearing


Blanchard, R., Morin, C., Malandrino, A., Vella, A., Sant, Z., Hellmich, C., (2016). Patient-specific fracture risk assessment of vertebrae: A multiscale approach coupling X-ray physics and continuum micromechanics International Journal for Numerical Methods in Biomedical Engineering , 32, (9), e02760

Summary: While in clinical settings, bone mineral density measured by computed tomography (CT) remains the key indicator for bone fracture risk, there is an ongoing quest for more engineering mechanics-based approaches for safety analyses of the skeleton. This calls for determination of suitable material properties from respective CT data, where the traditional approach consists of regression analyses between attenuation-related grey values and mechanical properties. We here present a physics-oriented approach, considering that elasticity and strength of bone tissue originate from the material microstructure and the mechanical properties of its elementary components. Firstly, we reconstruct the linear relation between the clinically accessible grey values making up a CT, and the X-ray attenuation coefficients quantifying the intensity losses from which the image is actually reconstructed. Therefore, we combine X-ray attenuation averaging at different length scales and over different tissues, with recently identified 'universal' composition characteristics of the latter. This gives access to both the normally non-disclosed X-ray energy employed in the CT-device and to in vivo patient-specific and location-specific bone composition variables, such as voxel-specific mass density, as well as collagen and mineral contents. The latter feed an experimentally validated multiscale elastoplastic model based on the hierarchical organization of bone. Corresponding elasticity maps across the organ enter a finite element simulation of a typical load case, and the resulting stress states are increased in a proportional fashion, so as to check the safety against ultimate material failure. In the young patient investigated, even normal physiological loading is probable to already imply plastic events associated with the hydrated mineral crystals in the bone ultrastructure, while the safety factor against failure is still as high as five.

JTD Keywords: Bone, Bone mass density, Continuum micromechanics, Elastoplasticity, Spine, Strength, X-ray physics


Aviles, A. I., Alsaleh, S., Montseny, E., Sobrevilla, P., Casals, A., (2016). A Deep-Neuro-Fuzzy approach for estimating the interaction forces in Robotic surgery FUZZ-IEEE IEEE International Conference on Fuzzy Systems , IEEE (Vancouver, Canada ) , 1113-1119

Fuzzy theory was motivated by the need to create human-like solutions that allow representing vagueness and uncertainty that exist in the real-world. These capabilities have been recently further enhanced by deep learning since it allows converting complex relation between data into knowledge. In this paper, we present a novel Deep-Neuro-Fuzzy strategy for unsupervised estimation of the interaction forces in Robotic Assisted Minimally Invasive scenarios. In our approach, the capability of Neuro-Fuzzy systems for handling visual uncertainty, as well as the inherent imprecision of real physical problems, is reinforced by the advantages provided by Deep Learning methods. Experiments conducted in a realistic setting have demonstrated the superior performance of the proposed approach over existing alternatives. More precisely, our method increased the accuracy of the force estimation and compared favorably to existing state of the art approaches, offering a percentage of improvement that ranges from about 35% to 85%.

JTD Keywords: Estimation, Force, Machine learning, Robots, Three-dimensional displays, Uncertainty, Visualization


Moles, E., Urbán, P., Jiménez-Díaz, M. B., Viera-Morilla, S., Angulo-Barturen, I., Busquets, M. A., Fernàndez-Busquets, X., (2015). Immunoliposome-mediated drug delivery to Plasmodium-infected and non-infected red blood cells as a dual therapeutic/prophylactic antimalarial strategy Journal of Controlled Release 210, 217-229

One of the most important factors behind resistance evolution in malaria is the failure to deliver sufficiently high amounts of drugs to early stages of Plasmodium-infected red blood cells (pRBCs). Despite having been considered for decades as a promising approach, the delivery of antimalarials encapsulated in immunoliposomes targeted to pRBCs has not progressed towards clinical applications, whereas in vitro assays rarely reach drug efficacy improvements above 10-fold. Here we show that encapsulation efficiencies reaching >96% are achieved for the weak basic drugs chloroquine (CQ) and primaquine using the pH gradient loading method in liposomes containing neutral saturated phospholipids. Targeting antibodies are best conjugated through their primary amino groups, adjusting chemical crosslinker concentration to retain significant antigen recognition. Antigens from non-parasitized RBCs have also been considered as targets for the delivery to the cell of drugs not affecting the erythrocytic metabolism. Using this strategy, we have achieved unprecedented complete nanocarrier targeting to early intraerythrocytic stages of the malaria parasite for which there is a lack of specific extracellular molecular tags. Immunoliposomes studded with monoclonal antibodies raised against the erythrocyte surface protein glycophorin A were capable of targeting 100% RBCs and pRBCs at the low concentration of 0.5 μM total lipid in the culture, with >95% of added liposomes retained on cell surfaces. When exposed for only 15 min to Plasmodium falciparum in vitro cultures of early stages, free CQ had no significant effect on the viability of the parasite up to 200 nM, whereas immunoliposomal 50 nM CQ completely arrested its growth. In vivo assays in mice showed that immunoliposomes cleared the pathogen below detectable levels at a CQ dose of 0.5 mg/kg, whereas free CQ administered at 1.75 mg/kg was, at most, 40-fold less efficient. Our data suggest that this significant improvement is in part due to a prophylactic effect of CQ found by the pathogen in its host cell right at the very moment of invasion.

JTD Keywords: Immunoliposomes, Malaria, Nanomedicine, Plasmodium, Targeted drug delivery


Ponce, I., Aragonès, A. C., Darwish, Nadrim, Pla-Vilanova, P., Oñate, R., Rezende, M. C., Zagal, J. H., Sanz, F., Pavez, J., Díez-Pérez, I., (2015). Building nanoscale molecular wires exploiting electrocatalytic interactions Electrochimica Acta 179, 611-167

Herein, we present a novel method to design nanoscale molecular wires by exploiting well-established electrocatalytic molecular platforms based on metallophthalocyanine blocks. Metallophthalocyanines exhibit high catalytic activity for a wide variety of electrochemical reactions of practical interests. To this aim, metallophthalocyanine molecules can be attached to an electrode surface via a conjugated mercaptopyridine axial ligand that provides (i) stable chemical binding to the metal surface through the thiol-anchoring group, and (ii) a good electrical communication between the metallophthalocyanine ring and the electrode surface. Our previous work demonstrates that long mercaptopyridinium blocks act as excellent linkers in such electrocatalytic platform, resulting in an optimal electrocatalytic activity of the metallophthalocyanine unit. Here we profit from this optimized electrocatalytic molecular platform to design new molecular wires that connect a metal nanoscale junction in a highly efficient and tunable way. To this aim, we use an STM break-junction approach to control the formation of a nanometric gap between two Au electrodes, both functionalized with mercaptopyridinium (bottom) and mercaptopyridine (top). When metallophthalocyanine is introduced into the functionalized metal nanojunction, stable molecular connections between the two electrodes are formed through axial coordination to the top and bottom pyridine moieties. We show that the highest conductance of the resulting nanoscale molecular wire corresponds to an Fe-phthalocyanine as compare to a Cu-phthalocyanine, which follows the electrocatalytic trend for such molecular systems. These results not only demonstrate a new strategy to design new families of highly conductive and tunable nanoscale molecular wires, but it also brings a new nanoscale electrical platform to help understanding some fundamental mechanistic aspects of molecular electrocatalysis.

JTD Keywords: Single-molecule wires, Metallophthalocyanine, Electrocatalytic molecular platform, Molecular Electronics, STM break-junction


Hoyo, J., Guaus, E., Torrent-Burgués, J., Sanz, F., (2015). Biomimetic monolayer films of digalactosyldiacylglycerol incorporating plastoquinone Biochimica et Biophysica Acta - Biomembranes , 1848, (6), 1341-1351

The photosynthesis is the process used by plants and bacteria cells to convert inorganic matter in organic thanks to the light energy. This process consist on several steps, being one of them the electronic transport from the photosystem II to the cytochrome thanks to plastoquinone-9 (PQ). Here we prepare membranes that mimic the characteristics and composition of natural photosynthetic cell membranes and we characterize them in order to obtain the PQ molecules position in the membrane and their electrochemical behaviour. The selected galactolipid is digalactosyldiacylglycerol (DGDG) that represents the 30% of the thylakoid membrane lipid content. The results obtained are worthful for several science fields due to the relevance of galactolipids as anti-algal, anti-viral, anti-tumor and anti-inflammatory agents and the antioxidant and free radical scavenger properties of prenylquinones. Both pure components (DGDG and PQ) and the DGDG:PQ mixtures have been studied using surface pressure-area isotherms. These isotherms give information about the film stability and indicate the thermodynamic behaviour of the mixture and their physical state. The Langmuir-Blodgett (LB) film has been transferred forming a monolayer that mimics the bottom layer of the biological membranes. This monolayer on mica has been topographically characterized using AFM and both the height and the physical state that they present have been obtained. Moreover, these monolayers have been transferred onto ITO that is a hydrophilic substrate with good optical and electrical features, so that, it is suitable for studying the electrochemical behaviour of these systems and it is a good candidate for energy producing devices.

JTD Keywords: Biomimetic membrane, Digalactosyldiacylglycerol, Electron transfer, LangmuirBlodgett film, Modified ITO electrode, Plastoquinone


Moles, E., Fernàndez-Busquets, X., (2015). Loading antimalarial drugs into noninfected red blood cells: An undesirable roommate for Plasmodium Future Medicinal Chemistry 7, (7), 837-840

The malaria parasite, Plasmodium spp., is a delicate unicellular organism unable to survive in free form for more than a couple of minutes in the bloodstream. Upon injection in a human by its Anopheles mosquito vector, Plasmodium sporozoites pass through the liver with the aim of invading hepatocytes. Those which succeed spend inside their host cell a recovery time before replicating and entering the blood circulation as fragile merozoites, although their exposure to host defenses is extraordinarily short. Quick invasion of red blood cells (RBCs) in a process lasting just a few minutes allows the parasite to escape immune system surveillance. For most of its erythrocytic cycle the pathogen feeds mainly on hemoglobin as it progresses from the early blood stages, termed rings, to the late forms trophozoites and schizonts. Early stages are ideal targets for antimalarial therapies because drugs delivered to them would have a longer time to kill the parasite before it completes its development. However, only 6 h after invasion does the permeability of the infected erythrocyte to anions and small nonelectrolytes, including some drugs, start to increase as the parasite matures [1]. During this maturation process the parasite hydrolyzes hemoglobin in a digestive vacuole, which is the target of many amphiphilic drugs that freely cross the RBC membrane and accumulate intracellularly. As a result, most antimalarials start affecting the infected cell relatively late in the intraerythrocytic parasite life cycle, when their effect is probably often too short to be lethal to Plasmodium.

JTD Keywords: Malaria, Nanomedicine, Plasmodium, Red blood cell, Targeted drug delivery


Aviles, A. I., Alsaleh, S., Sobrevilla, P., Casals, A., (2015). Sensorless force estimation using a neuro-vision-based approach for robotic-assisted surgery NER 2015 7th International IEEE/EMBS Conference on Neural Engineering , IEEE (Montpellier, France) , 86-89

This paper addresses the issue of lack of force feedback in robotic-assisted minimally invasive surgeries. Force is an important measure for surgeons in order to prevent intra-operative complications and tissue damage. Thus, an innovative neuro-vision based force estimation approach is proposed. Tissue surface displacement is first measured via minimization of an energy functional. A neuro approach is then used to establish a geometric-visual relation and estimate the applied force. The proposed approach eliminates the need of add-on sensors, carrying out biocompatibility studies and is applicable to tissues of any shape. Moreover, we provided an improvement from 15.14% to 56.16% over other approaches which demonstrate the potential of our proposal.

JTD Keywords: Estimation, Force, Minimally invasive surgery, Robot sensing systems, Three-dimensional displays


Pujol, A., Urbán, P., Riera, C., Fisa, R., Molina, I., Salvador, F., Estelrich, J., Fernàndez-Busquets, X., (2014). Application of quantum dots to the study of liposome targeting in leishmaniasis and malaria International Journal of Theoretical and Applied Nanotechnology , 2, (1), 1-8

Nanotechnological devices for therapeutic applications are massively addressed to diseases prevalent in the developed world, particularly cancer, because of the wrong assumption (for both ethical and technical reasons) that nanomedicines are too expensive and thus they can not be applied to diseases of poverty. Here we have applied quantum dots to study at the cellular level the delivery of the contents of liposomes to erythrocytes infected by the malaria parasite Plasmodium falciparum, and to macrophages infected by the leishmaniasis causative agent Leishmania infantum. A number of works have reported on the encapsulation in liposomes of drugs against both diseases as a strategy to increase therapeutic efficacy and decrease unspecific toxicity. Liposome-carried drugs end up inside Plasmodium-infected red blood cells (pRBCs) and in the phagolysosome system of Leishmania-infected macrophages but some knowledge gaps still obscure subcellular events related to these processes. As a proof of concept, we have used confocal fluorescence microscopy to follow the fate in pRBCs and infected macrophages of quantum dots encapsulated in liposomes, and of lysosomes, leishmaniasis and malaria parasites, nuclei, and phagosomes. Our data indicate that liposomes merge their lipid bilayers with pRBC plasma membranes but are engulfed by macrophages, where they fuse with lysosomes. Lysosomes have not been observed to join with phagosomes harboring single Leishmania parasites, whereas in phagosomes where the parasite has divided there is lysosome-specific fluorescence with a concomitant disappearance of lysosomes from the cytosol. In later stages, all the lysosome-specific label is found inside phagosomes whereas the phagosomal marker cadaverine strongly stains the macrophage nucleus, suggesting that Leishmania infection induces in its later stages nuclear degeneration and, possibly, apoptosis of the host cell. These results indicate that induction of macrophage apoptosis should be explored as a possible strategy used by Leishmania to prepare its egress.

JTD Keywords: Leishmania infantum, Leishmaniasis Liposomes, Malaria, Nanomedicine, Nanotechnology, Plasmodium falciparum, Quantum dots


Fernàndez-Busquets, X., (2014). Toy kit against malaria: Magic bullets, LEGO, Trojan horses and Russian dolls Therapeutic Delivery , 5, (10), 1049-1052

Movellan, J., Urbán, P., Moles, E., de la Fuente, J. M., Sierra, T., Serrano, J. L., Fernàndez-Busquets, X., (2014). Amphiphilic dendritic derivatives as nanocarriers for the targeted delivery of antimalarial drugs Biomaterials 35, (27), 7940-7950

It can be foreseen that in a future scenario of malaria eradication, a varied armamentarium will be required, including strategies for the targeted administration of antimalarial compounds. The development of nanovectors capable of encapsulating drugs and of delivering them to Plasmodium-infected cells with high specificity and efficacy and at an affordable cost is of particular interest. With this objective, dendritic derivatives based on 2,2-bis(hydroxymethyl)propionic acid (bis-MPA) and Pluronic® polymers have been herein explored. Four different dendritic derivatives have been tested for their capacity to encapsulate the antimalarial drugs chloroquine (CQ) and primaquine (PQ), their specific targeting to Plasmodium-infected red blood cells (pRBCs), and their antimalarial activity in vitro against the human pathogen Plasmodium falciparum and in vivo against the rodent malaria species Plasmodium yoelii. The results obtained have allowed the identification of two dendritic derivatives exhibiting specific targeting to pRBCs vs. non-infected RBCs, which reduce the in vitro IC50 of CQ and PQ by ca. 3- and 4-fold down to 4.0 nm and 1.1 μm, respectively. This work on the application of dendritic derivatives to antimalarial targeted drug delivery opens the way for the use of this new type of chemicals in future malaria eradication programs.

JTD Keywords: Antimalarial targeted drug delivery, Dendrimers, Malaria, Nanomedicine, Plasmodium, Polymeric nanoparticles


Urbán, P., Valle-Delgado, J. J., Mauro, N., Marques, J., Manfredi, A., Rottmann, M., Ranucci, E., Ferruti, P., Fernàndez-Busquets, X., (2014). Use of poly(amidoamine) drug conjugates for the delivery of antimalarials to Plasmodium Journal of Controlled Release 177, (1), 84-95

Current malaria therapeutics demands strategies able to selectively deliver drugs to Plasmodium-infected red blood cells (pRBCs) in order to limit the appearance of parasite resistance. Here, the poly(amidoamines) AGMA1 and ISA23 have been explored for the delivery of antimalarial drugs to pRBCs. AGMA1 has antimalarial activity per se as shown by its inhibition of the in vitrogrowth of Plasmodium falciparum, with an IC50 of 13.7 μM. Fluorescence-assisted cell sorting data and confocal fluorescence microscopy and transmission electron microscopy images indicate that both polymers exhibit preferential binding to and internalization into pRBCs versus RBCs, and subcellular targeting to the parasite itself in widely diverging species such as P. falciparum and Plasmodium yoelii, infecting humans and mice, respectively. AGMA1 and ISA23 polymers with hydrodynamic radii around 7 nm show a high loading capacity for the antimalarial drugs primaquine and chloroquine, with the final conjugate containing from 14.2% to 32.9% (w/w) active principle. Intraperitoneal administration of 0.8 mg/kg chloroquine as either AGMA1 or ISA23 salts cured P. yoelii–infected mice, whereas control animals treated with twice as much free drug did not survive. These polymers combining into a single chemical structure drug carrying capacity, low unspecific toxicity, high biodegradability and selective internalization into pRBCs, but not in healthy erythrocytes for human and rodent malarias, may be regarded as promising candidates deserving to enter the antimalarial therapeutic arena.

JTD Keywords: Malaria, Nanomedicine, Plasmodium, Polyamidoamines, Polymer-drug carriers, Targeted drug delivery


Pérez-Madrigal, M. M., Giannotti, M. I., Del Valle, L. J., Franco, L., Armelin, E., Puiggalí, J., Sanz, F., Alemán, C., (2014). Thermoplastic polyurethane:polythiophene nanomembranes for biomedical and biotechnological applications ACS Applied Materials & Interfaces 6, (12), 9719-9732

Nanomembranes have been prepared by spin-coating mixtures of a polythiophene (P3TMA) derivative and thermoplastic polyurethane (TPU) using 20:80, 40:60, and 60:40 TPU:P3TMA weight ratios. After structural, topographical, electrochemical, and thermal characterization, properties typically related with biomedical applications have been investigated: swelling, resistance to both hydrolytic and enzymatic degradation, biocompatibility, and adsorption of type I collagen, which is an extra cellular matrix protein that binds fibronectin favoring cell adhesion processes. The swelling ability and the hydrolytic and enzymatic degradability of TPU:P3TMA membranes increases with the concentration of P3TMA. Moreover, the degradation of the blends is considerably promoted by the presence of enzymes in the hydrolytic medium, TPU:P3TMA blends behaving as biodegradable materials. On the other hand, TPU:P3TMA nanomembranes behave as bioactive platforms stimulating cell adhesion and, especially, cell viability. Type I collagen adsorption largely depends on the substrate employed to support the nanomembrane, whereas it is practically independent of the chemical nature of the polymeric material used to fabricate the nanomembrane. However, detailed microscopy study of the morphology and topography of adsorbed collagen evidence the formation of different organizations, which range from fibrils to pseudoregular honeycomb networks depending on the composition of the nanomembrane that is in contact with the protein. Scaffolds made of electroactive TPU:P3TMA nanomembranes are potential candidates for tissue engineering biomedical applications.

JTD Keywords: Bioactive platform, Biodegradable blend, Collaged adsorption, Scaffolds, Tissue engineering, Ultrathin films


Navarro, S., Moleiro, V., Molina-Estevez, F. J., Lozano, M. L., Chinchon, R., Almarza, E., Quintana-Bustamante, O., Mostoslavsky, G., Maetzig, T., Galla, M., Heinz, N., Schiedlmeier, B., Torres, Y., Modlich, U., Samper, E., Río, P., Segovia, J. C., Raya, A., Güenechea, G., Izpisua-Belmonte, J. C., Bueren, J. A., (2014). Generation of iPSCs from genetically corrected Brca2 hypomorphic cells: Implications in cell reprogramming and stem cell therapy Stem Cells , 32, (2), 436-446

Fanconi anemia (FA) is a complex genetic disease associated with a defective DNA repair pathway known as the FA pathway. In contrast to many other FA proteins, BRCA2 participates downstream in this pathway and has a critical role in homology-directed recombination (HDR). In our current studies, we have observed an extremely low reprogramming efficiency in cells with a hypomorphic mutation in Brca2 (Brca2Δ27/Δ27), that was associated with increased apoptosis and defective generation of nuclear RAD51 foci during the reprogramming process. Gene complementation facilitated the generation of Brca2Δ27/Δ27 induced pluripotent stem cells (iPSCs) with a disease-free FA phenotype. Karyotype analyses and comparative genome hybridization arrays of complemented Brca2Δ27/Δ27 iPSCs showed, however, the presence of different genetic alterations in these cells, most of which were not evident in their parental Brca2 Δ27/Δ27 mouse embryonic fibroblasts. Gene-corrected Brca2Δ27/Δ27 iPSCs could be differentiated in vitro toward the hematopoietic lineage, although with a more limited efficacy than WT iPSCs or mouse embryonic stem cells, and did not engraft in irradiated Brca2Δ27/Δ27 recipients. Our results are consistent with previous studies proposing that HDR is critical for cell reprogramming and demonstrate that reprogramming defects characteristic of Brca2 mutant cells can be efficiently overcome by gene complementation. Finally, based on analysis of the phenotype, genetic stability, and hematopoietic differentiation potential of gene-corrected Brca2Δ27/Δ27 iPSCs, achievements and limitations in the application of current reprogramming approaches in hematopoietic stem cell therapy are also discussed.

JTD Keywords: Bone marrow aplasia, Cellular therapy, Fanconi anemia, Gene therapy, Hematopoietic stem cells, Induced pluripotent stem cells


Marques, J., Moles, E., Urbán, P., Prohens, R., Busquets, M. A., Sevrin, C., Grandfils, C., Fernàndez-Busquets, X., (2014). Application of heparin as a dual agent with antimalarial and liposome targeting activities toward Plasmodium-infected red blood cells Nanomedicine: Nanotechnology, Biology, and Medicine 10, (8), 1719-1728

Heparin had been demonstrated to have antimalarial activity and specific binding affinity for Plasmodium-infected red blood cells (pRBCs) vs. non-infected erythrocytes. Here we have explored if both properties could be joined into a drug delivery strategy where heparin would have a dual role as antimalarial and as a targeting element of drug-loaded nanoparticles. Confocal fluorescence and transmission electron microscopy data show that after 30. min of being added to living pRBCs fluorescein-labeled heparin colocalizes with the intracellular parasites. Heparin electrostatically adsorbed onto positively charged liposomes containing the cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane and loaded with the antimalarial drug primaquine was capable of increasing three-fold the activity of encapsulated drug in Plasmodium falciparum cultures. At concentrations below those inducing anticoagulation of mouse blood in vivo, parasiticidal activity was found to be the additive result of the separate activities of free heparin as antimalarial and of liposome-bound heparin as targeting element for encapsulated primaquine. From the Clinical Editor: Malaria remains an enormous global public health concern. In this study, a novel functionalized heparin formulation used as drug delivery agent for primaquine was demonstrated to result in threefold increased drug activity in cell cultures, and in a murine model it was able to provide these benefits in concentrations below what would be required for anticoagulation. Further studies are needed determine if this approach is applicable in the human disease as well.

JTD Keywords: Heparin, Liposomes, Malaria, Plasmodium, Targeted drug delivery, Heparin, Malaria, Plasmodium, Red blood cell, Targeted drug delivery, Liposomes, 1,2 dioleoyl 3 trimethylammoniopropane, fluorescein, heparin, liposome, nanoparticle, primaquine, adsorption, animal experiment, anticoagulation, antimalarial activity, Article, binding affinity, confocal microscopy, controlled study, drug targeting, encapsulation, erythrocyte, female, fluorescence microscopy, human, human cell, in vivo study, liposomal delivery, mouse, nonhuman, Plasmodium falciparum, transmission electron microscopy


Malandrino, Andrea, Lacroix, Damien, Hellmich, Christian, Ito, Keita, Ferguson, Stephen J., Noailly, J., (2014). The role of endplate poromechanical properties on the nutrient availability in the intervertebral disc Osteoarthritis and Cartilage , 22, (7), 1053-1060

Objective To investigate the relevance of the human vertebral endplate poromechanics on the fluid and metabolic transport from and to the intervertebral disc (IVD) based on educated estimations of the poromechanical parameter values of the bony endplate (BEP). Methods 50 micro-models of different BEP samples were generated from μCTs of lumbar vertebrae and allowed direct determination of porosity values. Permeability values were calculated by using the micro-models, through the simulation of permeation via computational fluid dynamics. These educated ranges of porosity and permeability values were used as inputs for mechano-transport simulations to assess their effect on both the distributions of metabolites within an IVD model and the poromechanical calculations within the cartilaginous part of the endplate i.e., the cartilage endplate (CEP). Results BEP effective permeability was highly correlated to local variations of porosity (R2 ≈ 0.88). Universal patterns between bone volume fraction and permeability arose from these results and from other experimental data in the literature. These variations in BEP permeability and porosity had negligible effects on the distributions of metabolites within the disc. In the CEP, the variability of the poromechanical properties of the BEP did not affect the predicted consolidation but induced higher fluid velocities. Conclusions The present paper provides the first sets of thoroughly identified BEP parameter values that can be further used in patient-specific poromechanical studies. Representing BEP structural changes through variations in poromechanical properties did not affect the diffusion of metabolites. However, attention might be paid to alterations in fluid velocities and cell mechano-sensing within the CEP.

JTD Keywords: Bony endplate, Spine mechanobiology, Intervertebral disc metabolites, Hydraulic Permeability, Bone Porosity, Poromechanics


Urbán, P., Fernàndez-Busquets, X., (2014). Nanomedicine against malaria Current Medicinal Chemistry , 21, (5), 605-629

Malaria is arguably one of the main medical concerns worldwide because of the numbers of people affected, the severity of the disease and the complexity of the life cycle of its causative agent, the protist Plasmodium sp. The clinical, social and economic burden of malaria has led for the last 100 years to several waves of serious efforts to reach its control and eventual eradication, without success to this day. With the advent of nanoscience, renewed hopes have appeared of finally obtaining the long sought-after magic bullet against malaria in the form of a nanovector for the targeted delivery of antimalarial drugs exclusively to Plasmodium-infected cells. Different types of encapsulating structure, targeting molecule, and antimalarial compound will be discussed for the assembly of Trojan horse nanocapsules capable of targeting with complete specificity diseased cells and of delivering inside them their antimalarial cargo with the objective of eliminating the parasite with a single dose. Nanotechnology can also be applied to the discovery of new antimalarials through single-molecule manipulation approaches for the identification of novel drugs targeting essential molecular components of the parasite. Finally, methods for the diagnosis of malaria can benefit from nanotools applied to the design of microfluidic-based devices for the accurate identification of the parasite's strain, its precise infective load, and the relative content of the different stages of its life cycle, whose knowledge is essential for the administration of adequate therapies. The benefits and drawbacks of these nanosystems will be considered in different possible scenarios, including cost-related issues that might be hampering the development of nanotechnology-based medicines against malaria with the dubious argument that they are too expensive to be used in developing areas.

JTD Keywords: Dendrimers, Liposomes, Malaria diagnosis, Nanobiosensors, Nanoparticles, Plasmodium, Polymers, Targeted drug delivery


Le Roux, D., Burger, P. B., Niemand, J., Grobler, A., Urbán, P., Fernàndez-Busquets, X., Barker, R. H., Serrano, A. E., I. Louw, A., Birkholtz, L. M., (2014). Novel S-adenosyl-L-methionine decarboxylase inhibitors as potent antiproliferative agents against intraerythrocytic Plasmodium falciparum parasites International Journal for Parasitology: Drugs and Drug Resistance , 4, (1), 28-36

S-adenosyl-l-methionine decarboxylase (AdoMetDC) in the polyamine biosynthesis pathway has been identified as a suitable drug target in Plasmodium falciparum parasites, which causes the most lethal form of malaria. Derivatives of an irreversible inhibitor of this enzyme, 5'-{[(Z)-4-amino-2-butenyl]methylamino}-5'-deoxyadenosine (MDL73811), have been developed with improved pharmacokinetic profiles and activity against related parasites, Trypanosoma brucei. Here, these derivatives were assayed for inhibition of AdoMetDC from P. falciparum parasites and the methylated derivative, 8-methyl-5'-{[(Z)-4-aminobut-2-enyl]methylamino}-5'-deoxyadenosine (Genz-644131) was shown to be the most active. The in vitro efficacy of Genz-644131 was markedly increased by nanoencapsulation in immunoliposomes, which specifically targeted intraerythrocytic P. falciparum parasites.

JTD Keywords: Immunoliposomes, Plasmodium, Polyamines, S-adenosyl-l-methionine decarboxylase


Juanola-Feliu, E., Miribel-Català, P. L., Avilés, C. P., Colomer-Farrarons, J., González-Piñero, M., Samitier, J., (2014). Design of a customized multipurpose nano-enabled implantable system for in-vivo theranostics Sensors 14, (10), 19275-19306

The first part of this paper reviews the current development and key issues on implantable multi-sensor devices for in vivo theranostics. Afterwards, the authors propose an innovative biomedical multisensory system for in vivo biomarker monitoring that could be suitable for customized theranostics applications. At this point, findings suggest that cross-cutting Key Enabling Technologies (KETs) could improve the overall performance of the system given that the convergence of technologies in nanotechnology, biotechnology, micro&nanoelectronics and advanced materials permit the development of new medical devices of small dimensions, using biocompatible materials, and embedding reliable and targeted biosensors, high speed data communication, and even energy autonomy. Therefore, this article deals with new research and market challenges of implantable sensor devices, from the point of view of the pervasive system, and time-to-market. The remote clinical monitoring approach introduced in this paper could be based on an array of biosensors to extract information from the patient. A key contribution of the authors is that the general architecture introduced in this paper would require minor modifications for the final customized bio-implantable medical device.

JTD Keywords: Biocompatible, Biosensor, Biotelemetry, Implantable multi-sensor, Innovation, KET, Nanomedicine, Personalized medicine, Biotelemetry, Innovation, Medical nanotechnology, Biocompatible, Implantable system, In-vivo, KET, Multi sensor, Personalized medicines, Theranostics, Biosensors


Mir, M., Lugo, R., Tahirbegi, I. B., Samitier, J., (2014). Miniaturizable ion-selective arrays based on highly stable polymer membranes for biomedical applications Sensors 14, (7), 11844-11854

Poly(vinylchloride) (PVC) is the most common polymer matrix used in the fabrication of ion-selective electrodes (ISEs). However, the surfaces of PVC-based sensors have been reported to show membrane instability. In an attempt to overcome this limitation, here we developed two alternative methods for the preparation of highly stable and robust ion-selective sensors. These platforms are based on the selective electropolymerization of poly(3,4-ethylenedioxythiophene) (PEDOT), where the sulfur atoms contained in the polymer covalently interact with the gold electrode, also permitting controlled selective attachment on a miniaturized electrode in an array format. This platform sensor was improved with the crosslinking of the membrane compounds with poly(ethyleneglycol) diglycidyl ether (PEG), thus also increasing the biocompatibility of the sensor. The resulting ISE membranes showed faster signal stabilization of the sensor response compared with that of the PVC matrix and also better reproducibility and stability, thus making these platforms highly suitable candidates for the manufacture of robust implantable sensors.

JTD Keywords: Biomedicine, Electrochemistry, Endoscope, Implantable device, Ion-selective electrode (ISE) sensor, Ischemia, pH detection, Biocompatibility, Chemical sensors, Electrochemistry, Electrodes, Electropolymerization, Endoscopy, Functional polymers, Implants (surgical), Ion selective electrodes, Medical applications, Polyvinyl chlorides, Stabilization, Biomedical applications, Biomedicine, Implantable devices, Ion selective sensors, Ischemia, Membrane instability, pH detection, Poly(3 ,4 ethylenedioxythiophene) (PEDOT), Ion selective membranes


Tahirbegi, I. B., Alvira, M., Mir, M., Samitier, J., (2014). Simple and fast method for fabrication of endoscopic implantable sensor arrays Sensors 14, (7), 11416-11426

Here we have developed a simple method for the fabrication of disposable implantable all-solid-state ion-selective electrodes (ISE) in an array format without using complex fabrication equipment or clean room facilities. The electrodes were designed in a needle shape instead of planar electrodes for a full contact with the tissue. The needle-shape platform comprises 12 metallic pins which were functionalized with conductive inks and ISE membranes. The modified microelectrodes were characterized with cyclic voltammetry, scanning electron microscope (SEM), and optical interferometry. The surface area and roughness factor of each microelectrode were determined and reproducible values were obtained for all the microelectrodes on the array. In this work, the microelectrodes were modified with membranes for the detection of pH and nitrate ions to prove the reliability of the fabricated sensor array platform adapted to an endoscope.

JTD Keywords: Chemical sensors, Cyclic voltammetry, Electrochemistry, Endoscopy, Fabrication, Implants (surgical), Microelectrodes, Needles, Nitrates, Scanning electron microscopy, Biomedicine, Fabricated sensors, Fabrication equipment, Implantable devices, Implantable sensors, Optical interferometry, Planar electrode, Roughness factor, Ion selective electrodes


Vaca, R., Aranda, J., (2014). Approximating coupler curves using strip trees Advanced Numerical Methods II 11th World Congress on Computational Mechanics (WCCM XI) 5th European Conference on Computational Mechanics (ECCM V) 6th European Conference on Computational Fluid Dynamics (ECFD VI) , CIMNE (Barcelona, Spain) , 1-2

For the mechanisms considered under the title linkages, coupler curve is the path traced by one of the point on the coupler link considered as an output of the mechanism which is joined to a fixed link. The equation of the coupler curve generated can be obtained solving a set of equations which describes distance constancy between all points of a mechanism and this coupler curve is the eliminant of these equations. The proposal to this work is to approximate coupler curves using strip trees.

JTD Keywords: Coupler curves, Strip tress, Distance geometry, Affine arithmetics, Planar linkages


Estrada, L., Torres, A., Garcia-Casado, J., Prats-Boluda, G., Yiyao, Ye-Lin, Jané, R., (2014). Evaluation of Laplacian diaphragm electromyographic recording in a dynamic inspiratory maneuver Engineering in Medicine and Biology Society (EMBC) 36th Annual International Conference of the IEEE , IEEE (Chicago, USA) , 2201-2204

The analysis of the electromyographic signal of the diaphragm muscle (EMGdi) can provide important information for evaluating the respiratory muscular function. The EMGdi can be recorded using surface Ag/AgCl disc electrodes in monopolar or bipolar configuration. However, these non-invasive EMGdi recordings are usually contaminated by the electrocardiographic (ECG) signal. EMGdi signal can also be noninvasively recorded using concentric ring electrodes in bipolar configuration (CRE) that estimate Laplacian surface potential. Laplacian recordings increase spatial resolution and attenuate distant bioelectric interferences, such as the ECG. Thus, the objective of this work is to compare and to evaluate CRE and traditional bipolar EMGdi recordings in a healthy subject during a dynamic inspiratory maneuver with incremental inspiratory loads. In the conducted study, it was calculated the cumulative percentage of power spectrum of EMGdi recordings to determine the signal bandwidth, and the power ratio between the EMGdi signal segments with and without cardiac activity. The results of this study suggest that EMGdi acquired with CRE electrodes is less affected by the ECG interference, achieves a wider bandwidth and a higher power ratio between segments without cardiac activity and with cardiac activity.

JTD Keywords: Bandwidth, Electric potential, Electrocardiography, Electrodes, Interference, Laplace equations, Muscles


Noailly, J., Malandrino, A., Galbusera, F., Jin, Zhongmin, (2014). Computational modelling of spinal implants Computational Modelling of Biomechanics and Biotribology in the Musculoskeletal System (ed. Jin, Z.), Woodhead Publishing (Cambridge, UK) Biomaterials and Tissues, 447-484

This chapter focuses on the use of the finite element method in the design and exploration of spinal implants. Following an introduction to biomechanical alterations of the spine in disease and to spine finite element modelling, focus is placed on different models developed for spine treatment simulations. Despite the hindrance of working thorough representations of in vivo situations, predictions of load transfer within both the implants and the tissues simulated allow improved interpretations of known clinical outcomes, and permit the educated design of new implants. The potential of probabilistic modelling is also discussed in relation to model validation and patient-specific analyses. Finally, the latest developments in the multiphysical modelling of intervertebral discs are presented, revealing a strong potential for the study of implant-based strategies that aim to restore the functional biophysics of the spine.

JTD Keywords: Spinal implant, Finite element modelling, Spine surgery, Spine biomechanics, Tissue mechanobiology


Estrada, L., Torres, A., Garcia-Casado, J., Ye-Lin, Y., Jané, R., (2014). Evaluation of Laplacian diaphragm electromyographic recordings in a static inspiratory maneuver IFMBE Proceedings XIII Mediterranean Conference on Medical and Biological Engineering and Computing 2013 (ed. Roa Romero, Laura M.), Springer International Publishing (London, UK) 41, 977-980

Diaphragm electromyography (EMGdi) provides important information on diaphragm activity, to detect neuromuscular disorders of the most important muscle in the breathing inspiratory phase. EMGdi is habitually recorded using needles or esophageal catheters, with the implication of being invasive for patients. Surface electrodes offer an alternative for the non-invasive assessment of diaphragm activity. Ag/AgCl surface disc electrodes are used in monopolar or bipolar configuration to record EMGdi signals. On the other hand, Laplacian surface potential can be estimated by signal recording through active concentric ring electrodes. This kind of recording could reduce physiological interferences, increase the spatial selectivity and reduce orientation problems in the electrode location. The aim of this work is to compare EMGdi signals recorded simultaneously with disc electrodes in bipolar configuration and a Laplacian ring electrode over chest wall. EMGdi signal was recorded in one healthy subject during a breath hold maneuver and a static inspiratory maneuver based on Mueller’s technique. In order to estimate the covered frequency range and the degree of noise contamination in both bipolar and Laplacian EMGdi signals, the cumulative percentage of the power spectrum and the signal to noise ratio in sub-bands were determined. Furthermore, diaphragm fatigue was evaluated by means of amplitude and frequency parameters. Our findings suggest that Laplacian EMGdi recording covers a broader frequency range although with higher noise contamination compared to bipolar EMGdi recording. Finally, in Laplacian recording fatigue indexes showed a clearer trend for muscle fatigue detection and also a reduced cardiac interference, providing an alternative to bipolar recording for diaphragm fatigue studies.

JTD Keywords: Laplacian electrode, Diaphragm muscle, Fatigue, Surface electromyography


Rajasekaran, V., Aranda, J., Casals, A., (2014). Handling disturbances on planned trajectories in robotic rehabilitation therapies IFMBE Proceedings XIII Mediterranean Conference on Medical and Biological Engineering and Computing 2013 (ed. Roa Romero, Laura M.), Springer International Publishing (London, UK) 41, 85-88

Robotic rehabilitation therapies are an emerging tool in the field of Neurorehabilitation in order to achieve an effective therapeutic development in the patient. In this paper, the role of disturbances caused by muscle synergies or unpredictable effects of artificial stimulation in muscles during rehabilitation therapies is analyzed. In terms of gait assistance it is also important to maintain synchronized movements to ensure a dynamically stable gait. Although, disturbances affecting joints are corrected by a force control approach, we define two methods to ensure stability and synchronization of joint movements in the trajectory to be followed. The performance of the presented methods is evaluated in comparison with a preplanned trajectory to be followed by the patients.

JTD Keywords: Exoskeleton, Force control, Gait assistance, Neurorobot, Trajectory planning


Juanola-Feliu, Esteve, Colomer-Farrarons, Jordi, Miribel-Català, Pere, González-Piñero, Manel, Samitier, Josep, (2014). Nano-enabled implantable device for glucose monitoring Implantable Bioelectronics (ed. Katz, Evgeny), Wiley-VCH Verlag GmbH & Co. KGaA (Weinheim, Germany) , 247-263

This chapter contains sections titled: * Introduction * Biomedical Devices for In Vivo Analysis * Conclusions and Final Recommendations * References

JTD Keywords: Technology transfer, Innovation management, Nanotechnology, Nanobiosensor, Diabetes, Biomedical device, Implantable biosensors


Fernàndez-Busquets, X., (2013). Amyloid fibrils in neurodegenerative diseases: villains or heroes? Future Medicinal Chemistry 5, (16), 1903-1906

Fernàndez-Busquets, X., (2013). Heparin-functionalized nanocapsules: Enabling targeted delivery of antimalarial drugs Future Medicinal Chemistry 5, (7), 737-739

Hoyo, J., Guaus, E., Oncins, G., Torrent-Burgués, J., Sanz, F., (2013). Incorporation of Ubiquinone in supported lipid bilayers on ITO Journal of Physical Chemistry B , 117, (25), 7498-7506

Ubiquinone (UQ) is one of the main electron and proton shuttle molecules in biological systems, and dipalmitoylphosphatidylcholine (DPPC) is one of the most used model lipids. Supported planar bilayers (SPBs) are extensively accepted as biological model membranes. In this study, SPBs have been deposited on ITO, which is a semiconductor with good electrical and optical features. Specifically, topographic atomic force microscopy (AFM) images and force curves have been performed on SPBs with several DPPC:UQ ratios to study the location and the interaction of UQ in the SPB. Additionally, cyclic voltammetry has been used to understand the electrochemical behavior of DPPC:UQ SPBs. Obtained results show that, in our case, UQ is placed in two main different positions in SPBs. First, between the DPPC hydrophobic chains, fact that originates a decrease in the breakthrough force of the bilayer, and the second between the two leaflets that form the SPBs. This second position occurs when increasing the UQ content, fact that eventually forms UQ aggregates at high concentrations. The formation of aggregates produces an expansion of the SPB average height and a bimodal distribution of the breakthrough force. The voltammetric response of UQ depends on its position on the bilayer.

JTD Keywords: Bimodal distribution, Biological models, Dipalmitoyl phosphatidylcholine, Electrochemical behaviors, Hydrophobic chains, Supported lipid bilayers, Supported planar bilayers, Voltammetric response


Pujol, A., Riera, C., Fisa, R., Molina, I., Salvador, F., Estelrich, J., Urbán, P., Fernàndez-Busquets, X., (2013). Nanomedicine for infectious diseases: Application of quantum dots encapsulated in immunoliposomes to the study of targeted drug delivery against leishmaniasis and malaria Proceedings of the 4th International Conference on Nanotechnology: Fundamentals and Applications. 4th International Conference on Nanotechnology: Fundamentals and Applications , International ASET Inc. (Ontario, Canada) , 1-8

Nanotechnological devices for therapeutic applications are massively addressed to diseases prevalent in the developed world, particularly cancer, because of the wrong assumption (for both ethical and technical reasons) that nanomedicines are too expensive and thus they can not be applied to diseases of poverty. Here we have applied quantum dots to study at the cellular level the delivery of the contents of immunoliposomes to erythrocytes infected by the malaria parasite Plasmodium falciparum, and to macrophages infected by the leishmaniasis causative agent Leishmania infantum. A number of works have reported on the encapsulation in liposomes of drugs against both diseases as a strategy to increase therapeutic efficacy and decrease unspecific toxicity. Liposome-carried drugs end up inside Plasmodium-infected red blood cells (pRBCs) and in the phagolysosome system of Leishmania-infected macrophages but some knowledge gaps still obscure subcellular events related to these processes. As a proof of concept, we have used confocal fluorescence microscopy to follow the fate in pRBCs and L. infantum-infected macrophages of quantum dots encapsulated in liposomes, and of lysosomes, Leishmania and Plasmodium parasites, nuclei, and phagosomes. Our data indicate that liposomes merge their lipid bilayers with pRBC plasma membranes but are engulfed by macrophages, where they fuse with lysosomes. Lysosomes have not been observed to join with phagosomes harboring single L. infantum parasites, whereas in phagosomes where the parasite has divided there is lysosome-specific fluorescence with a concomitant disappearance of lysosomes from the cytosol. In later stages, all the lysosome-specific label is found inside phagosomes whereas the phagosomal marker cadaverine strongly stains the macrophage nucleus, suggesting that L. infantum infection induces in its later stages nuclear degeneration and possibly, apoptosis of the host cell. These results indicate that induction of macrophage apoptosis should be explored as a possible strategy used by L. infantum to prepare its egress.

JTD Keywords: Leishmania infantum, Leishmaniasis, Liposomes, Malaria, Nanomedicine, Nanotechnology, Plasmodium falciparum, Quantum dots


Gilbert, M., Juárez, A., Madrid, C., Balsalobre, C., (2013). New insights in the role of HtdA in the regulation of R27 conjugation Plasmid International Society for Plasmid Biology Meeting , Elsevier (Santander, Spain) 70 (1), 61-68

R27 is the prototype of the IncHI group of conjugative plasmids, which are associated with multidrug resistance in several relevant pathogens. The transfer of this plasmid is thermodependent and all transfer-related genes are encoded in six operons (tra operons). Very little is known about the factors involved in the regulation of the R27 conjugation. This report describes transcriptional studies of the six tra operons. Our results indicate that HtdA, encoded in the R27 plasmid, is involved in the transcriptional repression of four tra operons (F, H, AC and Z). Although HtdA plays a pivotal role in the transcriptional regulation of those operons, it does not exert its effect as a classical repressor. The data indicate the existence of a crosstalk between HtdA and other unknown regulatory factors. The HtdA-mediated regulation of conjugation is independent of the R27 H-NS protein.

JTD Keywords: Plasmid conjugation, IncHI, R27, tra Operons regulation, HtdA


Paytubia, S., Dietrich, M., Queiroz, M.H., Juárez, A., (2013). Role of plasmid- and chromosomally encoded Hha proteins in modulation of gene expression in E. coli O157:H7 Plasmid International Society for Plasmid Biology Meeting , Elsevier (Santander, Spain) 70 (1), 52-60

H-NS and Hha belong to the nucleoid-associated family of proteins and modulate gene expression in response to environmental stimuli. Genes coding for these proteins can be either chromosomally or plasmid-encoded. In this work, we analyse the regulatory role of the Hha protein encoded in the virulence plasmid of the enterohemorrhagic Escherichia coli O157:H7 (HhapO157). This plasmid is present in all clinical isolates of E. coli O157:H7 and contributes to virulence. Both, HhapO157 and E. coli O157:H7-chromosomal Hha (Hhachr) exhibit a significant degree of similarity. The hha gene from plasmid pO157 is transcribed from its own putative promoter and is overexpressed in a chromosomal hha mutant. As its chromosomal counterpart, HhapO157 is able to interact with H-NS. Remarkably, HhapO157 targets only a subset of the genes modulated by Hhachr. This has been evidenced by both assaying the ability of HhapO157 to complement expression of a specific operon (i.e., the haemolysin operon) and by comparing the global transcriptome of the wt strain and its hhap, hhac and hhapc mutant derivatives. HhapO157 and Hhachr share some common regulatory features, however they also display specific targeting of some genes and even a different modulatory role in some others.

JTD Keywords: E. coli O157:H7, Hha, H-NS, Plasmid, pO157, Nucleoid-associated proteins


Esteban, O., Christ, D., Stock, D., (2013). Purification of molecular machines and nanomotors using phage-derived monoclonal antibody fragments Protein Nanotechnology - Methods in Molecular Biology (ed. Gerrard, J. A.), Humana Press (New York, USA) 996, 203-217

Molecular machines and nanomotors are sophisticated biological assemblies that convert potential energy stored either in transmembrane ion gradients or in ATP into kinetic energy. Studying these highly dynamic biological devices by X-ray crystallography is challenging, as they are difficult to produce, purify, and crystallize. Phage display technology allows us to put a handle on these molecules in the form of highly specific antibody fragments that can also stabilize conformations and allow versatile labelling for electron microscopy, immunohistochemistry, and biophysics experiments. Here, we describe a widely applicable protocol for selecting high-affinity monoclonal antibody fragments against a complex molecular machine, the A-type ATPase from T. thermophilus that allows fast and simple purification of this transmembrane rotary motor from its wild-type source. The approach can be readily extended to other integral membrane proteins and protein complexes as well as to soluble molecular machines and nanomotors.

JTD Keywords: ATP synthase, Crystallization, Domain antibodies, Electron microscopy, Labelling, Membrane proteins, Monoclonal antibody fragments, Phage display, Protein purification, X-ray crystallography


Mir, Mònica , Tahirbegi, Islam Bogachan , Valle-Delgado, Juan José , Fernàndez-Busquets, X., Samitier, Josep , (2012). In vitro study of magnetite-amyloid β complex formation Nanomedicine: Nanotechnology, Biology, and Medicine 8, (6), 974-980

Biogenic magnetite (Fe3O4) has been identified in human brain tissue. However, abnormal concentration of magnetite nanoparticles in the brain has been observed in different neurodegenerative pathologies. In the case of Alzheimer's disease (AD), these magnetic nanoparticles have been identified attached to the characteristic brain plaques, which are mainly formed by fibrils of amyloid β peptide (Aβ). However, few clues about the formation of the magnetite-Aβ complex have been reported. We have investigated the interaction between these important players in the AD with superconducting quantum interference, scanning electron microscope, surface plasmon resonance, and magnetic force microscopy. The results support the notion that the magnetite-Aβ complex is created before the synthesis of the magnetic nanoparticles, bringing a highly stable interaction of this couple.

JTD Keywords: Alzheimer's disease, Biogenic magnetite, Amyloid β peptide (Aβ), Superconducting quantum interference, Scanning electron microscope, Surface plasmon resonance, Magnetic force microscopy


Tort, N., Salvador, J. P., Avino, A., Eritja, R., Comelles, J., Martinez, E., Samitier, J., Marco, M. P., (2012). Synthesis of steroid-oligonucleotide conjugates for a DNA site-encoded SPR immunosensor Bioconjugate Chemistry , 23, (11), 2183-2191

The excellent self-assembling properties of DNA and the excellent specificity of the antibodies to detect analytes of small molecular weight under competitive conditions have been combined in this study. Three oligonucleotide sequences (N(1)up, N(2)up, and N(3)up) have been covalently attached to three steroidal haptens (8, hG, and 13) of three anabolic-androgenic steroids (AAS), stanozolol (ST), tetrahydrogestrinone (THG), and boldenone (B), respectively. The synthesis of steroid oligonucleotide conjugates has been performed by the reaction of oligonucleotides carrying amino groups with carboxyl acid derivatives of steroidal haptens. Due to the chemical nature of the steroid derivatives, two methods for coupling the haptens and the ssDNA have been studied: a solid-phase coupling strategy and a solution-phase coupling strategy. Specific antibodies against ST, THG, and B have been used in this study to asses the possibility of using the self-assembling properties of the DNA to prepare biofunctional SPR gold chips based on the immobilization of haptens, by hybridization with the complementary oligonucleotide strands possessing SH groups previously immobilized. The capture of the steroid oligonucleotide conjugates and subsequent binding of the specific antibodies can be monitored on the sensogram due to variations produced on the refractive index on top of the gold chip. The resulting steroid oligonucleotide conjugates retain the hybridization and specific binding properties of oligonucleotides and haptens as demonstrated by thermal denaturation experiments and surface plasmon resonance (SPR).

JTD Keywords: Directed protein immobilization, Plasmon resonance biosensor, Self-assembled monolayers, Label-free, Serum samples, Assay, Immunoassays, Antibodies, Progress, Binding


Juanola-Feliu, E., Colomer-Farrarons, J., Miribel-Català , P., Samitier, J., Valls-Pasola, J., (2012). Market challenges facing academic research in commercializing nano-enabled implantable devices for in-vivo biomedical analysis Technovation , 32, (3-4), 193-204

This article reports on the research and development of a cutting-edge biomedical device for continuous in-vivo glucose monitoring. This entirely public-funded process of technological innovation has been conducted at the University of Barcelona within a context of converging technologies involving the fields of medicine, physics, chemistry, biology, telecommunications, electronics and energy. The authors examine the value chain and the market challenges faced by in-vivo implantable biomedical devices based on nanotechnologies. In so doing, they trace the process from the point of applied research to the final integration and commercialization of the product, when the social rate of return from academic research can be estimated. Using a case-study approach, the paper also examines the high-tech activities involved in the development of this nano-enabled device and describes the technology and innovation management process within the value chain conducted in a University-Hospital-Industry-Administration-Citizens framework. Here, nanotechnology is seen to represent a new industrial revolution, boosting the biomedical devices market. Nanosensors may well provide the tools required for investigating biological processes at the cellular level in vivo when embedded into medical devices of small dimensions, using biocompatible materials, and requiring reliable and targeted biosensors, high speed data transfer, safely stored data, and even energy autonomy.

JTD Keywords: Biomedical device, Diabetes, Innovation management, Nanobiosensor, Nanotechnology, Research commercialization, Technology transfer, Academic research, Applied research, Barcelona, Biocompatible materials, Biological process, Biomedical analysis, Biomedical devices, Cellular levels, Converging technologies, Glucose monitoring, High-speed data transfer, Implantable biomedical devices, Implantable devices, In-vivo, Industrial revolutions, Innovation management, Medical Devices, Nanobiosensor, Rate of return, Research and development, Technological innovation, Value chains, Biological materials, Biomedical engineering, Biosensors, Commerce, Data transfer, Earnings, Engineering education, Glucose, Implants (surgical), Industrial research, Innovation, Medical problems, Nanosensors, Nanotechnology, Technology transfer, Equipment


Pegueroles, M., Tonda-Turo, C., Planell, J. A., Gil, F. J., Aparicio, C., (2012). Adsorption of fibronectin, fibrinogen, and albumin on TiO2: Time-resolved kinetics, structural changes, and competition study Biointerphases , 7, (48), 13

An understanding of protein adsorption process is crucial for designing biomaterial surfaces. In this work, with the use of a quartz-crystal microbalance with dissipation monitoring, we researched the following: (a) the kinetics of adsorption on TiO2 surfaces of three extensively described proteins that are relevant for metallic implant integration [i.e., albumin (BSA), fibrinogen (Fbg), and fibronectin (Fn)]; and (b) the competition of those proteins for adsorbing on TiO2 in a two-step experiment consisted of sequentially exposing the surfaces to different monoprotein solutions. Each protein showed a different process of adsorption and properties of the adlayer-calculated using the Voigt model. The competition experiments showed that BSA displaced larger proteins such as Fn and Fbg when BSA was introduced as the second protein in the system, whereas the larger proteins laid on top of BSA forming an adsorbed protein bi-layer when those were introduced secondly in the system.

JTD Keywords: QCM, Human plasma fibronectin, Induced conformational-changes, Von-willebrand-factor, BSA, Protein adsortion, Polymer surfaces, Solid-surfaces, Viscoelastic properties, Globular-proteins


Simao, C., Mas-Torrent, M., Crivillers, N., Lloveras, V., Artés, Juan Manuel, Gorostiza, Pau, Veciana, Jaume, Rovira, C., (2011). A robust molecular platform for non-volatile memory devices with optical and magnetic responses Nature Chemistry , 3, (5), 359-364

Bistable molecules that behave as switches in solution have long been known. Systems that can be reversibly converted between two stable states that differ in their physical properties are particularly attractive in the development of memory devices when immobilized in substrates. Here, we report a highly robust surface-confined switch based on an electroactive, persistent organic radical immobilized on indium tin oxide substrates that can be electrochemically and reversibly converted to the anion form. This molecular bistable system behaves as an extremely robust redox switch in which an electrical input is transduced into optical as well as magnetic outputs under ambient conditions. The fact that this molecular surface switch, operating at very low voltages, can be patterned and addressed locally, and also has exceptionally high long-term stability and excellent reversibility and reproducibility, makes it a very promising platform for non-volatile memory devices.

JTD Keywords: Self-assembled monolayers, Chromophore-based monolayers, Ultrathin platinum films, Carbon free-radicals, Per-million levels, Polychlorotriphenylmethyl radicals, Electron-transfer, Surface, Logic, Quantification


Carulla, Patricia, Bribian, Ana, Rangel, Alejandra, Gavin, Rosalina, Ferrer, Isidro, Caelles, Carme, Antonio del Rio, Jose, Llorens, Franc, (2011). Neuroprotective role of PrP(C) against kainate-induced epileptic seizures and cell death depends on the modulation of JNK3 activation by GluR6/7-PSD-95 binding Molecular Biology of the Cell , 22, (17), 3041-3054

Cellular prion protein (PrP(C)) is a glycosyl-phosphatidylinositol-anchored glycoprotein. When mutated or misfolded, the pathogenic form (PrP(SC)) induces transmissible spongiform encephalopathies. In contrast, PrP(C) has a number of physiological functions in several neural processes. Several lines of evidence implicate PrP(C) in synaptic transmission and neuroprotection since its absence results in an increase in neuronal excitability and enhanced excitotoxicity in vitro and in vivo. Furthermore, PrP(C) has been implicated in the inhibition of N-methyl-D-aspartic acid (NMDA)-mediated neurotransmission, and prion protein gene (Prnp) knockout mice show enhanced neuronal death in response to NMDA and kainate (KA). In this study, we demonstrate that neurotoxicity induced by KA in Prnp knockout mice depends on the c-Jun N-terminal kinase 3 (JNK3) pathway since Prnp(%) Jnk3(%) mice were not affected by KA. Pharmacological blockage of JNK3 activity impaired PrP(C)-dependent neurotoxicity. Furthermore, our results indicate that JNK3 activation depends on the interaction of PrP(C) with postsynaptic density 95 protein (PSD-95) and glutamate receptor 6/7 (GluR6/7). Indeed, GluR6-PSD-95 interaction after KA injections was favored by the absence of PrP(C). Finally, neurotoxicity in Prnp knockout mice was reversed by an AMPA/KA inhibitor (6,7-dinitroquinoxaline-2,3-dione) and the GluR6 antagonist NS-102. We conclude that the protection afforded by PrP(C) against KA is due to its ability to modulate GluR6/7-mediated neurotransmission and hence JNK3 activation.

JTD Keywords: Ischemic brain-injury, Prion protein PrP(C), Stress-inducible protein-1, Synaptic plasticity, Neurite outgrowth, Signaling module, Caspase-3 activation, Organotypic cultures, Cerebral-ischemia


Urban, Patricia, Estelrich, Joan, Adeva, Alberto, Cortes, Alfred, Fernàndez-Busquets, X., (2011). Study of the efficacy of antimalarial drugs delivered inside targeted immunoliposomal nanovectors Nanoscale Research Letters 6, (1), 620

Paul Ehrlich's dream of a 'magic bullet' that would specifically destroy invading microbes is now a major aspect of clinical medicine. However, a century later, the implementation of this medical holy grail continues being a challenge in three main fronts: identifying the right molecular or cellular targets for a particular disease, having a drug that is effective against it, and finding a strategy for the efficient delivery of sufficient amounts of the drug in an active state exclusively to the selected targets. In a previous work, we engineered an immunoliposomal nanovector for the targeted delivery of its contents exclusively to Plasmodium falciparum-infected red blood cells [pRBCs]. In preliminary assays, the antimalarial drug chloroquine showed improved efficacy when delivered inside immunoliposomes targeted with the pRBC-specific monoclonal antibody BM1234. Because difficulties in determining the exact concentration of the drug due to its low amounts prevented an accurate estimation of the nanovector performance, here, we have developed an HPLC-based method for the precise determination of the concentrations in the liposomal preparations of chloroquine and of a second antimalarial drug, fosmidomycin. The results obtained indicate that immunoliposome encapsulation of chloroquine and fosmidomycin improves by tenfold the efficacy of antimalarial drugs. The targeting antibody used binds preferentially to pRBCs containing late maturation stages of the parasite. In accordance with this observation, the best performing immunoliposomes are those added to Plasmodium cultures having a larger number of late form-containing pRBCs. An average of five antibody molecules per liposome significantly improves in cell cultures the performance of immunoliposomes over non-functionalized liposomes as drug delivery vessels. Increasing the number of antibodies on the liposome surface correspondingly increases performance, with a reduction of 50% parasitemia achieved with immunoliposomes encapsulating 4 nM chloroquine and bearing an estimated 250 BM1234 units. The nanovector prototype described here can be a valuable platform amenable to modification and improvement with the objective of designing a nanostructure adequate to enter the preclinical pipeline as a new antimalarial therapy.

JTD Keywords: Plasmodium falciparum, Antimalarial drug, Nanovector, Immuno-liposomes


Roa, J. J., Oncins, G., Diaz, J., Capdevila, X. G., Sanz, F., Segarra, M., (2011). Study of the friction, adhesion and mechanical properties of single crystals, ceramics and ceramic coatings by AFM Journal of the European Ceramic Society 31, (4), 429-449

This paper reviews commonly used methods of analyzing and interpreting friction, adhesion and nanoindentation with an AFM tip test data, with a particular emphasis of the testing of single crystals, metals, ceramics and ceramic coatings. Experimental results are reported on the friction, mechanical and adhesion properties of these materials. The popularity of AFM testing is evidenced by the large quantity of papers that report such measurements in the last decade. Unfortunately, a lot of information about these topics is scare in the literature. The present paper is aimed to present the basic physical modelling employed and also some examples using each technique.

JTD Keywords: Mechanical properties, Plasticity, Biomedical applications, Engine components


Hristova, K., Pecheva, E., Pramatarova, L., Altankov, G., (2011). Improved interaction of osteoblast-like cells with apatite-nanodiamond coatings depends on fibronectin Journal of Materials Science: Materials in Medicine , 22, (8), 1891-1900

New apatite (AP)/nanodiamond (ND) coating has been developed to improve physical and biological properties of stainless steel (SS) versus single AP coating. Homogeneously electrodeposited AP-ND layer demonstrates increased mechanical strength, interlayer cohesion and ductility. In the absence of serum, osteoblast-like MG63 cells attach well but poorly spread on both AP and AP-ND substrata. Pre-adsorption with serum or fibronectin (FN) improves the cellular interaction-an effect that is better pronounced on the AP-ND coating. In single protein adsorption study fluorescein isothiocyanate-labeled FN (FITC-FN) shows enhanced deposition on the AP-ND layer consistent with the significantly improved cell adhesion, spreading and focal adhesions formation (in comparison to SS and AP), particularly at low FN adsorption concentrations (1 mu g/ml). Higher FN concentrations (20 mu g/ml) abolish this difference suggesting that the promoted cellular interaction of serum (where FN is low) is caused by the greater affinity for FN. Moreover, it is found that MG63 cells tend to rearrange both adsorbed and secreted FN on the AP-ND layer suggesting facilitated FN matrix formation.

JTD Keywords: Extracellular-matrix, Protein adsorption, Integrins, Adhesion, Biomaterials, Surfaces, Polymerization, Composite, Implants, Titanium


Morgenstern, R., Morgenstern, C., Jané, R., Lee, S. H., (2011). Usefulness of an expandable interbody spacer for the treatment of foraminal stenosis in extremely collapsed disks preliminary clinical experience with endoscopic posterolateral transforaminal approach Journal of Spinal Disorders & Techniques , 24, (8), 485-491

Study Design: Clinical series of patients with degenerative disk disease undergoing an endoscopic posterolateral transforaminal procedure that used a reaming foraminoplasty technique to enlarge the foramen coupled with insertion of the B-Twin expandable spacer. Objectives: This retrospective analysis of 107 consecutive patients sought to assess the outcome of this surgical procedure. Summary of Background Data: Reamed endoscopic foraminoplasty under direct endoscopic vision has been shown to be suitable for extremely collapsed disks (> 50% total disk height) despite the difficult access, especially at L5-S1. The authors tried to investigate the efficacy of an expandable spacer being inserted by the endoscopic transforaminal approach to solve foraminal stenosis without bone fusion techniques. Methods: The procedure consists of bone reaming under direct endoscopic control to wide the foramen followed by insertion of the B-Twin expandable device as a disk spacer to restore partially or to maintain the height of the collapsed disk. Outcome measures included visual analog scale (VAS) for pain, the Oswestry Disability Index (ODI) for functional disability, and radioimaging studies. Results: Mean follow-up was 27.2 months. Clinical outcome was considered excellent in 64 patients, good in 25, fair in 10, and poor in 8. Results were similar in single and double B-Twin spacer insertions. Postoperative mean values for VAS and ODI scores improved significantly as compared with preoperative data. Mean VAS and ODI scores were significantly higher in patients with fair or poor results than in those with excellent or good outcome. In 2 cases, clear signs of end plate bone resorption in the control computed tomographic scans at 6 months and 12 months leading to a substantial loss of disk height were documented. Conclusions: This preliminary study has shown the efficacy of an endoscopic surgical technique for the treatment of foraminal stenosis in extremely collapsed disks.

JTD Keywords: Foraminal stenosis, B-twin expandable spacer, Endoscopic foraminoplasty, Minimally invasive surgery, Surgical technique, Spinal spacer, Lumbar, Diskectomy, Fusion, Discectomy


Colomer-Farrarons, Jordi , Miribel-Català, Pedro Luís, Samitier, Josep , (2011). Low-voltage µpower CMOS subcutaneous biomedical implantable device for true/false applications Biomedical Engineering IASTED International Conference Biomedical Engineering (Biomed 2011) (ed. Baumgartner, C.), ACTA Press (Innsbruck, Austria) Biomedical Engineering, 424-428

A ±1.2V / 350μW integrated front-end architecture for a true/false in-vivo subcutaneous detection device is presented. The detection is focused on using three electrodes amperometric sensors. The powering and AM transcutaneous communication are based on an inductively coupled link working at 13.56 MHz. A prototype device (5.5 mm x 29.5 mm) has been implemented and fully validated.

JTD Keywords: Implantable Device, Front-End architecture, Bioelectronics, Microelectronics Design, Biosensors


Garcia-Manyes, S., Redondo-Morata, L., Oncins, G., Sanz, F., (2010). Nanomechanics of lipid bilayers: Heads or tails? Journal of the American Chemical Society American Chemical Society 132, (37), 12874-12886

Understanding the effect of mechanical stress on membranes is of primary importance in biophysics. Here we use force spectroscopy AFM to quantitatively characterize the nanomechanical stability of supported lipid bilayers as a function of their chemical composition. The onset of plastic deformation reveals itself as a repetitive jump in the approaching force curve, which represents a molecular fingerprint for the bilayer mechanical stability. By systematically probing a set of chemically distinct supported lipid bilayers (SLBs), we first show that both the headgroup and tail have a decisive effect on their mechanical properties. While the mechanical stability of the probed SLBs linearly increases by 3.3 nN upon the introduction of each additional -CH2- in the chain, it exhibits a significant dependence on the phospholipid headgroup, ranging from 3 nN for DPPA to 66 nN for DPPG. Furthermore, we also quantify the reduction of the membrane mechanical stability as a function of the number of unsaturations and molecular branching in the chemical structure of the apolar tails. Finally, we demonstrate that, upon introduction of cholesterol and ergosterol, contrary to previous belief the mechanical stability of membranes not only increases linearly in the liquid phase (DLPC) but also for phospholipids present in the gel phase (DPPC). Our results are discussed in the framework of the continuum nucleation model. This work highlights the compelling effect of subtle variations in the chemical structure of phospholipid molecules on the membrane response when exposed to mechanical forces, a mechanism of common occurrence in nature.

JTD Keywords: Atomic-force microscopy, Molecular-dynamics simulation, Aqueous-electrolyte solutions, Supported planar membranes, Phospholipid-bilayers, Biological-membranes, Physical-properties, Fluid membranes, Model membranes, Chain-length


Gil, Vanessa, Bichler, Zoe, Lee, Jae K., Seira, Oscar, Llorens, Franc, Bribian, Ana, Morales, Ricardo, Claverol-Tinture, Enric, Soriano, Eduardo, Sumoy, Lauro, Zheng, Binhai, del Rio, Jose A., (2010). Developmental expression of the oligodendrocyte myelin glycoprotein in the mouse telencephalon Cerebral Cortex , 20, (8), 1769-1779

The oligodendrocyte myelin glycoprotein is a glycosylphosphatidylinositol-anchored protein expressed by neurons and oligodendrocytes in the central nervous system. Attempts have been made to identify the functions of the myelin-associated inhibitory proteins (MAIPs) after axonal lesion or in neurodegeneration. However, the developmental roles of some of these proteins and their receptors remain elusive. Recent studies indicate that NgR1 and the recently discovered receptor PirB restrict cortical synaptic plasticity. However, the putative factors that trigger these effects are unknown. Because Nogo-A is mostly associated with the endoplasmic reticulum and myelin associated glycoprotein appears late during development, the putative participation of OMgp should be considered. Here, we examine the pattern of development of OMgp immunoreactive elements during mouse telencephalic development. OMgp immunoreactivity in the developing cortex follows the establishment of the thalamo-cortical barrel field. At the cellular level, we located OMgp neuronal membranes in dendrites and axons as well as in brain synaptosome fractions and axon varicosities. Lastly, the analysis of the barrel field in OMgp-deficient mice revealed that although thalamo-cortical connections were formed, their targeting in layer IV was altered, and numerous axons ectopically invaded layers II-III. Our data support the idea that early expressed MAIPs play an active role during development and point to OMgp participating in thalamo-cortical connections.

JTD Keywords: Axon plasticity, Barrel-field specification, Cortical lamination, Myelin


Gavín, R., Ferrer, Isidro, del Río, J. A., (2010). Involvement of Dab1 in APP processing and [beta]-amyloid deposition in sporadic Creutzfeldt-Jakob patients Neurobiology of Disease , 37, (2), 324-329

Alzheimer's disease and prion pathologies (e.g., Creutzfeldt-Jakob disease (CJD)) display profound neural lesions associated with aberrant protein processing and extracellular amyloid deposits. Dab1 has been implicated in the regulation of amyloid precursor protein (APP), but a direct link between human prion diseases and Dab1/APP interactions has not been published. Here we examined this putative relationship in 17 cases of sporadic CJD (sCJD) post-mortem. Biochemical analyses of brain tissue revealed two groups, which also correlated with PrPsc types 1 and 2. One group with PrPsc type 1 showed increased Dab1 phosphorylation and lower [beta]CTF production with an absence of A[beta] deposition. The second sCJD group, which carried PrPsc type 2, showed lower levels of Dab1 phosphorylation and [beta]CTF production, and A[beta] deposition. Thus, the present observations suggest a correlation between Dab1 phosphorylation, A[beta] deposition and PrPsc type in sCJD.

JTD Keywords: Prionopathies, Amyloid plaques, Alzheimer's disease, Dab1


Cervera, M., Esteban, O., Gil, M., Gorris, M. T., Martínez, M. C., Peña, L., Cambra, M., (2010). Transgenic expression in citrus of single-chain antibody fragments specific to Citrus tristeza virus confers virus resistance Transgenic Research , 19, (6), 1001-1015

Citrus tristeza virus (CTV) causes one of the most destructive viral diseases of citrus worldwide. Generation of resistant citrus genotypes through genetic engineering could be a good alternative to control CTV. To study whether production of single-chain variable fragment (scFv) antibodies in citrus could interfere and immunomodulate CTV infection, transgenic Mexican lime plants expressing two different scFv constructs, separately and simultaneously, were generated. These constructs derived from the well-referenced monoclonal antibodies 3DF1 and 3CA5, specific against CTV p25 major coat protein, whose mixture is able to detect all CTV isolates characterized so far. ScFv accumulation levels were low and could be readily detected just in four transgenic lines. Twelve homogeneous and vigorous lines were propagated and CTV-challenged by graft inoculation with an aggressive CTV strain. A clear protective effect was observed in most transgenic lines, which showed resistance in up to 40-60% of propagations. Besides, both a delay in symptom appearance and attenuation of symptom intensity were observed in infected transgenic plants compared with control plants. This effect was more evident in lines carrying the 3DF1scFv transgene, being probably related to the biological functions of the epitope recognized by this antibody. This is the first report describing successful protection against a pathogen in woody transgenic plants by ectopic expression of scFv recombinant antibodies.

JTD Keywords: CTV control, Immunomodulation, Plantibodies, Recombinant antibodies, Transgenic citrus


Banos, R. C., Vivero, A., Aznar, S., Garcia, J., Pons, M., Madrid, C., Juarez, A., (2009). Differential regulation of horizontally acquired and core genome genes by the bacterial modulator H-NS PLoS Genetics 5, (6), 8

Horizontal acquisition of DNA by bacteria dramatically increases genetic diversity and hence successful bacterial colonization of several niches, including the human host. A relevant issue is how this newly acquired DNA interacts and integrates in the regulatory networks of the bacterial cell. The global modulator H-NS targets both core genome and HGT genes and silences gene expression in response to external stimuli such as osmolarity and temperature. Here we provide evidence that H-NS discriminates and differentially modulates core and HGT DNA. As an example of this, plasmid R27-encoded H-NS protein has evolved to selectively silence HGT genes and does not interfere with core genome regulation. In turn, differential regulation of both gene lineages by resident chromosomal H-NS requires a helper protein: the Hha protein. Tight silencing of HGT DNA is accomplished by H-NS-Hha complexes. In contrast, core genes are modulated by H-NS homoligomers. Remarkably, the presence of Hha-like proteins is restricted to the Enterobacteriaceae. In addition, conjugative plasmids encoding H-NS variants have hitherto been isolated only from members of the family. Thus, the H-NS system in enteric bacteria presents unique evolutionary features. The capacity to selectively discriminate between core and HGT DNA may help to maintain horizontally transmitted DNA in silent form and may give these bacteria a competitive advantage in adapting to new environments, including host colonization.

JTD Keywords: 2A strain 2457T, Escherichia-Coli, Salmonella-Enterica, Protein, DNA, Expression, Binding, HHA, Shigella, Plasmid


Zhou, E. H., Trepat, X., Park, C. Y., Lenormand, G., Oliver, M. N., Mijailovich, S. M., Hardin, C., Weitz, D. A., Butler, J. P., Fredberg, J. J., (2009). Universal behavior of the osmotically compressed cell and its analogy to the colloidal glass transition Proceedings of the National Academy of Sciences of the United States of America 106, (26), 10632-10637

Mechanical robustness of the cell under different modes of stress and deformation is essential to its survival and function. Under tension, mechanical rigidity is provided by the cytoskeletal network; with increasing stress, this network stiffens, providing increased resistance to deformation. However, a cell must also resist compression, which will inevitably occur whenever cell volume is decreased during such biologically important processes as anhydrobiosis and apoptosis. Under compression, individual filaments can buckle, thereby reducing the stiffness and weakening the cytoskeletal network. However, the intracellular space is crowded with macromolecules and organelles that can resist compression. A simple picture describing their behavior is that of colloidal particles; colloids exhibit a sharp increase in viscosity with increasing volume fraction, ultimately undergoing a glass transition and becoming a solid. We investigate the consequences of these 2 competing effects and show that as a cell is compressed by hyperosmotic stress it becomes progressively more rigid. Although this stiffening behavior depends somewhat on cell type, starting conditions, molecular motors, and cytoskeletal contributions, its dependence on solid volume fraction is exponential in every instance. This universal behavior suggests that compression-induced weakening of the network is overwhelmed by crowding-induced stiffening of the cytoplasm. We also show that compression dramatically slows intracellular relaxation processes. The increase in stiffness, combined with the slowing of relaxation processes, is reminiscent of a glass transition of colloidal suspensions, but only when comprised of deformable particles. Our work provides a means to probe the physical nature of the cytoplasm under compression, and leads to results that are universal across cell type.

JTD Keywords: Compression, Cytoplasm, Cytoskeleton, Mechanotransduction, Stiffness


Barhoumi, H., Haddad, R., Maaref, A., Bausells, J., Bessueille, F., Leonard, D., Jaffrezic-Renault, N., Martelet, C., Zine, N., Errachid, A., (2008). Na+-implanted membrane for a capacitive sodium electrolyte-Insulator-Semiconductor microsensors Sensor Letters International Conference of Thermal, Mechanical and Multiphysics Simulation and Experiments in Microelectronics and Microsystems (ed. -----), Amer Scientific Publishers (Lombardy, Italy) 6, (1), 204-208

Ion implanted Insulator-Semiconductor (IS) sensor that specifically detects Na+ ions have been developed using ion implantation technique. Na+ ions were directly implanted with ion energies 30, 45, and 60 keV into the IS (oxidized Si3N4/Si3N4/SiO2/P-Si) structures previously covered with a thin aluminum layer. X-ray photoelectron spectroscopy (XPS) characterization shows that sodium and aluminum ions were implanted into the oxidized Si3N4 insulating layer surface. Their atomic percentage depending on energy, fluence of the implanted ion and of the annealing temperature. The sen sitivity of the ion-implanted IS structure for Na+ and of some interfering (K+, Li+, H+, and NH4+) ions was investigated using high frequency capacitance-voltage measurements. Under optimal i mplantation conditions such as energy, fluence and annealing temperature, the developed sodium microsensor demonstrates quasi-nernstian sensitivity (50 +/- 2 mV/pNa) in the concentration range from 10(-3.7) to 10(-1) M and high lifetime greater than 16 months without any loss of sensitivity.

JTD Keywords: Na+ microsensor, Ion implantation, XPS, C-V measurements


Domènech, Ò., Morros, A., Cabañas, M. E., Teresa Montero, M., Hernéndez-Borrell, J., (2007). Supported planar bilayers from hexagonal phases Biochimica et Biophysica Acta - Biomembranes , 1768, (1), 100-106

In this work the presence of inverted hexagonal phases HII of 1-palmitoy-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and cardiolipin (CL) (0.8:0.2, mol/mol) in the presence of Ca2+ were observed via 31P-NMR spectroscopy. When suspensions of the same composition were extended onto mica, HII phases transformed into structures which features are those of supported planar bilayers (SPBs). When characterized by atomic force microscopy (AFM), the SPBs revealed the existence of two laterally segregated domains (the interdomain height being ∼ 1 nm). Cytochrome c (cyt c), which binds preferentially to acidic phospholipids like CL, was used to demonstrate the nature of the domains. We used 1-anilinonaphtalen-8-sulfonate (ANS) to demonstrate that in the presence of cyt c, the fluorescence of ANS decreased significantly in lamellar phases. Conversely, the ANS binding to HII phases was negligible. When cyt c was injected into AFM fluid imaging cells, where SPBs of POPE:CL had previously formed poorly defined structures, protein aggregates (∼ 100 nm diameter) were ostensibly observed only on the upper domains, which suggests not only that they are mainly formed by CL, but also provides evidence of bilayer formation from HII phases. Furthermore, a model for the nanostructure of the SPBs is herein proposed.

JTD Keywords: 31P-NMR, AFM, ANS fluorescence, Cytochrome c (cyt c), Hexagonal phase (HII), Liposome, Supported planar bilayers (SPBs)