A comparative investigation of Bombyx mori silk fibroin hydrogels generated by chemical and enzymatic cross-linking.

Fibroin, the major proteinaceous component of the silk fiber produced by larvae of the domesticated silk moth (Bombyx mori), has been widely investigated as a biomaterial for potential applications in tissue engineering and regenerative medicine. Following sol-gel transition, silk fibroin solutions can generate hydrogels that present certain advantages when employed as biomaterials, especially if they are cross-linked. The subject of this study was the self-cross-linking of silk fibroin through a process induced by the enzyme horseradish peroxidase (HRP) in the presence of hydrogen peroxide, a method only recently proposed and scarcely reported. The hydrogels were prepared either by physical cross-linking, by cross-linking with a natural compound (genipin), or by enzymatic cross-linking. The products were comparatively characterized in regard to their synthesis and background chemical aspects, physical and optical properties, mechanical properties, secondary structure, swelling/deswelling behavior, enzymatic degradation, and compatibility as substrates for cell adhesion and proliferation. The study confirmed the advantages of the HRP-induced cross-linking, which included considerably shorter gelation times, enhanced elasticity of the resulting hydrogels, and improved cytocompatibility. Discrepancies between certain results of this investigation and those reported previously were discussed in detail.