The electrospinning technique allows engineering biomimetic scaffolds within micro to nanoscale range mimicking natural extracellular matrix (ECM). Chitosan (CS) and polycaprolactone (PCL) were dissolved in a modified solvent mixture consisting of formic acid and acetone (3:7) and mixed in different weight ratios to get chitosan-polycaprolactone [CS-PCL] blend solutions. The CS-PCL blend polymer was electrospun in the same solvent system and compared with PCL. The physicochemical characterization of the electrospun fibrous mats was done using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), tensile test, swelling properties, water contact angle (WCA) analysis, surface profilometry and thermo gravimetric analysis (TGA). The CS-PCL fibrous mat showed decreased hydrophobicity. The CS-PCL mats also showed improved swelling property, tensile strength, thermal stability and surface roughness. The cytocompatibility of the CS-PCL and PCL fibrous mats were examined using mouse fibroblast (L-929) cell line by direct contact and cellular activity with extract of materials confirmed non-cytotoxic nature. The potential of CS-PCL and PCL fibrous mats as skin tissue engineering scaffolds were assessed by cell adhesion, viability, proliferation and actin distribution using human keratinocytes (HaCaT) and L-929 cell lines. Results indicate that CS-PCL is a better scaffold for attachment and proliferation of keratinocytes and is a potential material for skin tissue engineering.
The conventional method of retrieving cells for tissue engineering to create three-dimensional functional tissues uses enzymes that may hamper cell viability and re-adhesion. Culturing cells on thermoresponsive surfaces of poly(N-isopropylacrylamide) (PNIPAAm) is a relatively new nondestructive method of creating in vitro tissues. In this study, PNIPAAm and glycidylmethacrylate (GMA)-based thermoresponsive copolymer N-isopropylacrylamide-co-glycidylmethacrylate (NGMA) were synthesized as a potential cell culture harvesting system for generating 3D synthetic tissues. The copolymer was characterized by differential scanning calorimetry, gel permeation chromatography, Fourier transform infrared spectroscopy, water contact angle, atomic force microscopy, and nuclear magnetic resonance spectroscopy. The NGMA-coated dishes were evaluated for cytotoxicity and cytocompatibility using L-929 cells. Primary rabbit corneal cultures established on NGMA surface were detached as an intact cell sheet with epithelial specific characteristics as well as maintenance of cell—cell and cell—extracellular matrix contact. The results confirmed the suitability of NGMA substrate for cell culture and temperature-induced cell sheet harvest. This is the first report on this copolymer formulation as a substrate for tissue engineering application. Hydrophobic GMA apart from modulating the lower critical solution temperature features the prospects of further modification, namely the incorporation of biomolecules through the epoxy groups.
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