Preparation and in vitro characterization of cross-linked collagen–gelatin hydrogel using EDC/NHS for corneal tissue engineering applications

Corneal disease is considered as the second leading cause of vision loss and keratoplasty is known as an effective treatment for it. However, the tissue engineered corneal substitutes are promising tools in experimental in vivo repair of cornea. Selecting appropriate cell sources and scaffolds are two important concerns in corneal tissue engineering. The object of this study was to investigate biocompatibility and physical properties of the bio-engineered cornea, fabricated from type-I collagen (COL) and gelatin (Gel). Two gelatin based hydrogels cross-linked with EDC/NHS were fabricated, and their physicochemical properties such as equilibrium water content, enzymatic degradation, mechanical properties, rheological, contact angle and optical properties as well as their ability to support human bone-marrow mesenchymal stem cells (hBM-MSCs) survival were characterized. The equilibrium water content and enzymatic degradation of these hydrogels can be easily controlled by adding COL. Our findings suggest that incorporation of COL-I increases optical properties, hydrophilicity, stiffness and Young's modulus. The viability of hBM-MSCs cultured in Gel and Gel: COL was assessed via CCK-8 assay. Also, the morphology of the hBM-MSCs on the top of Gel and Gel: COL hydrogels were characterized by phase-contrast microscopy. This biocompatible hydrogel may promise to be used as artificial corneal substitutes.