摘要

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.