Decellularized bovine pericardium (DBP)-based biomeshes are the gold standard in reconstructive surgery. In order to prolong their stability after the transplantation, various chemical cross-linking strategies are employed. However, structural and functional properties of the biomeshes differ in dependence on the cross-linker used. Here, we performed a bottom-up study of structural and functional alterations of DBP-based biomeshes following cross-linking with hexamethylene diisocyanate (HMDC), ethylene glycol diglycidyl ether (EGDE), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and genipin. The in vitro cytotoxicity tests supported their clinical applicability. Their structural differences (eg roughness, fibre thickness, pore morphology) were evaluated using the two-photon confocal laser scanning, atomic force, scanning electron and polarized light microscopies. HMDC and EDC samples appeared to be the roughest. Complex mechanical trials indicated the tendency to reduced Young's Modulus and mechanical anisotropy values of DBP upon cross-linking. The lowest mechanical anisotropy was found in EDC and genipin sample groups.In vitro collagenase susceptibility was the highest for EDC samples and the lowest for EGDE samples. The comparative analysis of the results allowed us to recognize the strengths and weaknesses of each cross-linker in relation to a particular clinical application.
K E Y W O R D Sbiomechanics, bovine pericardium, cross-linking, decellularization, reconstructive surgery
Over the last decade, a number of hydrogels attracted great attention in the area of brain tissue engineering. The hydrogels are composed of hydrophilic polymers forming 3D network in water. Their function is promoting structural and functional restoration of damaged brain tissues by providing mechanical support and navigating cell fate. This paper reports on the neurocompatibility of chitosan-g-oligo(L,L-lactide) copolymer hydrogel with primary rat cortical neuron culture. The hydrogel was produced by a molding technique on the base of photocurable composition consisting of chitosan-g-oligo(L,L-lactide) copolymer, poly(ethylene glycol) diacrylate and photosensitizer Irgacure 2959. The influence of the hydrogel on cell viability, phenotype and calcium homeostasis, mitochondrial potential and oxygen consumption rate in glutamate excitotoxicity was analyzed using primary neuron cultures obtained from a neonatal rat cortex. This study revealed that the hydrogel is non-cytotoxic. Dissociated neonatal rat cortical cells were actively attaching to the hydrogel surface and exhibited the phenotype, calcium homeostasis and mitochondrial function in both standard conditions and glutamate excitotoxicity (100 μM) similar to the control cells cultured without the hydrogel. To conclude, in this study we assessed the feasibility of the application of chitosan-g-oligo(L,L-lactide) copolymer hydrogel for tissue engineering therapy of brain injury in an in vitro model. The results support that the hydrogel is able to sustain realization of the functional metabolic activity of neonatal rat cortical cells in response to glutamate excitotoxicity.
Enhancement of cell adhesion and growth on surface of the biodegradable materials is one of the important tasks in development of materials for regenerative medicine. This work focuses on comparison of various methods of collagen coating deposition onto polylactide films, aiming to increase their biocompatibility with human mesenchymal stromal cells. The collagen deposition was realized using either preliminary plasma treatment of the polylactide films or pre-swelling in solvent mixture. These techniques were compared in terms of the effect on the surface’s chemical structure, morphology, hydrophilicity and ability to support adhesion and growth of human mesenchymal stromal cells.
Chitosan (CS)/graphene nanocomposite films with tunable biomechanics, electroconductivity and biocompatibility using polyvinylpyrrolidone (PVP) and Pluronic F108 (Plu) as emulsion stabilizers for the purpose of conductive tissue engineering were successfully obtained. In order to obtain a composite solution, aqueous dispersions of multilayered graphene stabilized with Plu/PVP were supplied with CS at a ratio of CS to stabilizers of 2:1, respectively. Electroconductive films were obtained by the solution casting method. The electrical conductivity, mechanical properties and in vitro and in vivo biocompatibility of the resulting films were assessed in relation to the graphene concentration and stabilizer type and they were close to that of smooth muscle tissue. According to the results of the in vitro cytotoxicity analysis, the films did not release soluble cytotoxic components into the cell culture medium. The high adhesion of murine fibroblasts to the films indicated the absence of contact cytotoxicity. In subcutaneous implantation in Wistar rats, we found that stabilizers reduced the brittleness of the chitosan films and the inflammatory response.
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