Fibroin/collagen hybrid hydrogels with crosslinking method: Preparation, properties, and cytocompatibility

Substrate stiffness is an important physical factor in the response of many cell types. Although protein-based hydrogels are widely used as cell-culture substrates because of their resemblance to the natural extracellular matrix (ECM) and complex signaling, their rigidity should be further increased to facilitate the adhesion and growth of cells. In this study, fibroin/collagen hydrogels having suitable stiffness were prepared directly by adding 1-ethyl-3-(3-dimethylaminoprophy) carbodiimide hydrochloride (EDC) in fibroin/collagen solution to induce crosslinking. The storage moduli of these crosslinked hydrogels are above 3 kPa, and even exceed 10 kPa, having stronger mechanical strength than that of previously reported protein-based hydrogels. Furthermore, the crosslinked hydrogels can maintain their configuration above 80(o)C, which proves their increased thermal stability. Although crosslinked, the hydrogels still maintain the mobility of fibroin molecules. The growth of vascular smooth muscle cells (VSMCs) in fibroin/collagen gels indicates that the crosslinking reaction has no negative influence on the biocompatibility of fibroin/collagen hydrogels. The fibroin/collagen hydrogels are more propitious to the growth of cells compared with fibroin/collagen scaffolds. Because of their inherent biocompatibility, excellent mechanical and thermal properties, and green preparation process, the fibroin/collagen hydrogels would become promising scaffolds for tissue engineering.