Silk fibroin hydrogel membranes prepared by a sequential cross-linking strategy for guided bone regeneration

Silk fibroin hydrogel is a widely used material for tissue engineering. However, its mechanical properties are the main obstacle to its application to medical bone regeneration barrier membranes. Here, we developed a new hydrogel membrane for guided bone regeneration (GBR). In this study, a sequential crosslinking strategy including photo crosslinking and organic solvent (ethanol) treatment was used to obtain silk fibroin hydrogel membrane (EA-SF). The mechanical properties of EA-SF were significantly enhanced compared to the hydrogel prepared only by photocrosslinking (E-SF). The compressive and tensile strengths of the hydrogel film treated with 75% ethanol for 6 h were 1.18 ± 0.36 MPa and 0.43 ± 0.03 MPa, respectively. In vitro cell culture results showed that EA-SF has good biocompatibility and can effectively shield fibroblasts (L929). EA-SF also has excellent in vitro protein hydrolysis stability. In vivo experiments (subcutaneous implantation and calvarial defect experiment) confirmed the stability and barrier functionality of EA-SF. In conclusion, this study demonstrated that ethanol could improve the mechanical properties of silk fibroin hydrogel and extend the scope of their application, making silk fibroin hydrogel a promising GBR material.