Enhancing Cell Seeding and Osteogenesis of MSCs on 3D Printed Scaffolds Through Injectable BMP2 Immobilized ECM-Mimetic Gel

Objective Design of bioactive scaffolds with osteogenic capacity is a central challenge in cell-based patient-specific bone tissue engineering. Efficient and spatially uniform seeding of (stem) cells onto such constructs is vital to attain functional tissues. Herein we developed heparin functionalized collagen gels supported by 3D printed bioceramic scaffolds, as bone extracellular matrix (ECM)-mimetic matrices. These matrices were designed to enhance cell seeding efficiency of mesenchymal stem cells (MSCs) as well as improve their osteogenic differentiation through immobilized bone morphogenic protein 2 (BMP2) to be used for personalized bone regeneration. Methods A 3D gel based on heparin-conjugated collagen matrix capable of immobilizing recombinant human bone morphogenic protein 2 (BMP2) was synthesized. Isolated dental pulp Mesenchymal stem cells (MSCs) were then encapsulated into the bone ECM microenvironment to efficiently and uniformly seed a bioactive ceramic-based scaffold fabricated using additive manufacturing technique. The designed 3D cell-laden constructs were comprehensively investigated trough in vitro assays and in vivo study. Results In-depth rheological characterizations of heparin-conjugated collagen gel revealed that elasticity of the matrix is significantly improved compared with freely incorporated heparin. Investigation of the MSCs laden collagen-heparin hydrogels revealed their capability to provide spatiotemporal bioavailability of BMP2 while suppressing the matrix contraction over time. The in vivo histology and real-time polymerase chain reaction (qPCR) analysis showed that the designed construct supported the osteogenic differentiation of MSCs and induced the ectopic bone formation in rat model. Significance The presented hybrid constructs combine bone ECM chemical cues with mechanical function providing an ideal 3D microenvironment for patient-specific bone tissue engineering and cell therapy applications. The implemented methodology in design of ECM-mimetic 3D matrix capable of immobilizing BMP2 to improve seeding efficiency of customized scaffolds can be exploited for other bioactive molecules

Montmorillonite modified by CNx supported Pt for methanol oxidation

A composite support based on nature clay, i.e. montmorillonite (MMT), shows great promise as support materials for Pt electrocatalyst for the methanol oxidation reaction in fuel cell anodes. The reported composite support (CNx-MMT) was prepared via carbonizing MMT which was covered by N-contented polymer. X-ray diffraction and transmission electron microscopy results showed that Pt nanoparticles can be well-dispersed on the composite support with highly dispersed tiny crystal Pt nanoparticles. Cyclic voltammetry measurements showed that the Pt/CNx-MMT has the enhanced electrocatalytic activity in methanol oxidation reaction. The developed Pt catalyst supported on new composite support is catalytically more active for methanol electrooxidation than Pt supported on the conventional carbon support and shows good stability, offering promising potential for application of MMT as support for fuel cell electrocatalysis.Web of Scienc

Versatile Sarcosine Biosensing Schemes Utilizing Layer-by-Layer Construction of Carbon Nanotube-Chitosan Composite Films

Layer-by-layer composite films of carbon nanotubes (CNTs) within a chitosan matrix with sarcosine oxidase enzyme and capped with Nafion have been developed and optimized as a versatile 1st generation amperometric sarcosine biosensing platform that operates successfully both as an isolated sarcosine sensor as well as a functional component within a creatinine sensor. Accurate measurement of sarcosine in urine and creatinine in blood may help with early diagnosis of diseases such as prostate cancer and renal failure, respectively. In this study, each material within the film is systematically optimized toward sarcosine sensitivity, including a critical evaluation of different CNTs effect on sensing performance. Films featuring carboxylic acid–modified single–walled carbon nanotubes and strategic enzyme doping were shown to be most effective sarcosine sensing platforms, exhibiting excellent sensitivity (~0.5 μA/mM), a linear response (≤ 0.75 mM), fast response time (8 s), low limits of detection (~6 μM), as well as both continuous use stability (7 days) and effective shelf life (\u3e 12 days). Operation of the sarcosine sensor was demonstrated in a urine matrix, detecting sarcosine at physiologically relevant concentrations and successfully quantifying sarcosine-spiked urine samples with high percent recovery and low relative error. The sarcosine sensing platform was also adapted to a 1st generation creatinine biosensing scheme in which the sarcosine enzymatic reaction is critical to a trienzymatic cascade event. The creatinine sensor yielded sensitivity of ~0.6 μA/mM, similar sensing performance parameters to the sarcosine sensor, and was effectively operated in blood serum at physiologically relevant creatinine concentrations. The demonstrated functionality of these sensors in their respective biological fluids at physiological concentrations of the analyte species suggests potential clinical application as diagnostic tools

Nanocomposite SPEEK-based membranes for Direct Methanol Fuel Cells at intermediate temperatures

Novel nanocomposite membranes were prepared by infiltration of a blend of sulfonated PEEK (SPEEK) with polyvinyl alcohol (PVA), using water as solvent, into electrospun nanolibers of SPEEK blended with polyvinyl butyral (PVB). The membranes were characterized for their application on Direct Methanol Fuel Cells (DMFCs) operating at moderate temperatures (>80 degrees C). An important role of the solvent on the crosslinking temperature for the SPEEK-PVA system was observed. A mat of hydrated SPEEK-30%PVB nanofibers revealed higher proton conductivity in comparison with a dense membrane of similar composition. Incorporation of the nanoliber mats to the SPEEK-35%PVA matrix provided mechanical stability, methanol barrier properties and certain proton conductivity up to a crosslinking temperature of 120 degrees C. Not remarkable effect of the nanofibers was found above that crosslinking temperature. The combined effect of the nanofibers and crosslinking temperature on the properties of the membranes is discussed. DIV1FC performance experiments concluded promising results for this new low-cost type of membranes, although further optimization steps are still required.This research has been funded by the R&D Support Programmes of the Polytechnic University of Valencia (project 24761) and the Spanish Ministry of Science and Innovation (project SP-ENE-20120718).Mollá Romano, S.; Compañ Moreno, V. (2015). Nanocomposite SPEEK-based membranes for Direct Methanol Fuel Cells at intermediate temperatures. Journal of Membrane Science. 492:123-136. https://doi.org/10.1016/j.memsci.2015.05.055S12313649

Chitosan: poly (vinyl) alcohol composite alkaline membrane incorporating organic ionomers and layered silicate materials into a PEM electrochemical reactor

Mixed matrix membranes (MMM) are prepared from equivalent blends of poly (vinyl alcohol) (PVA) and chitosan (CS) polymers doped with organic ionomers 4VP and AS4, or inorganic layered titanosilicate AM-4 and stannosilicate UZAR-S3, by solution casting to improve the mechanical and thermal properties, hydroxide conductivity and alcohol barrier effect to reduce the crossover. The structural properties, thermal stability, hydrolytic stability, transport and ionic properties of the prepared composite membranes were investigated by scanning electron microscopy (SEM), X-ray diffraction (XRD), thermogravimetric analysis (TGA), water uptake, water content, alcohol permeability, thickness, ion exchange capacity (IEC) and OH- conductivity measurements. The addition of both organic and inorganic fillers in a CS:PVA blend polymer enhances the thermal and ionic properties. All the membranes are homogenous, as revealed by the SEM and XRD studies, except when UZAR-S3 stannosilicate is used as filler, which leads to a dual layer structure, a top layer of UZAR-S3 lamellar particles bound together by the polymer matrix and a bottom layer composed mostly of polymer blend. The loss of crystallinity was especially remarkable in 4VP/CS:PVA membrane. Thus, the 4VP/CS:PVA membrane exhibits the best ionic conductivity, whereas the UZAR-S3/CS:PVA membrane the best reduced alcohol crossover. Finally, the performance of the CS:PVA-based membranes were tested in a Polymer Electrolyte Membrane Electrochemical Reactor (PEMER) for the feasibility use of alkaline anionic exchange membranes in electrosynthesis under alkaline conditions, showing the 4VP/CS:PVA and UZAR-S3/CS:PVA membranes the best performances in PEMER.We gratefully acknowledge the financial support from the Spanish Ministry of Economy and Competitiveness (MINECO) for CTQ2012-31229 project at the University of Cantabria, and MINECO-FEDER (Fondo Europeo de Desarrollo Regional (2014- 2020) through the CTQ2013-48280-C3-3-R project at the University of Alicante. C. C. C. also thanks the MINECO for the “Ramón y Cajal” program at the University of Cantabria (RYC2011-08550), and L. G. C. for her PhD fellowship BES-2011-045147 and the EEBB- 14-09094 mobility grant for the research stay at the University of Cantabria, respectively. Dr. César Rubio, Prof. Carlos Téllez and Prof. Joaquín Coronas from the University of Zaragoza are also warmly thanked for the UZAR-S3 sample