Nanosilicate‐Reinforced Silk Fibroin Hydrogel for Endogenous Regeneration of Both Cartilage and Subchondral Bone

Abstract Osteochondral defects are characterized by injuries to both cartilage and subchondral bone, which is a result of trauma, inflammation, or inappropriate loading. Due to the unique biological properties of subchondral bone and cartilage, developing a tissue engineering scaffold that can promote dual‐lineage regeneration of cartilage and bone simultaneously remains a great challenge. In this study, a microporous nanosilicate‐reinforced enzymatically crosslinked silk fibroin (SF) hydrogel is fabricated by introducing montmorillonite (MMT) nanoparticles via intercalation chemistry. In vitro studies show that SF‐MMT nanocomposite hydrogel has improved mechanical properties and hydrophilicity, as well as the bioactivities to promote the osteogenic differentiation of bone marrow mesenchymal stem cells and maintain chondrocyte phenotype compared with SF hydrogel. Global proteomic analysis verifies the dual‐lineage bioactivities of SF‐MMT nanocomposite hydrogel, which are probably regulated by multiple signaling pathways. Furthermore, it is observed that the biophysical interaction of cells and SF‐MMT nanocomposite hydrogel is partially mediated by clathrin‐mediated endocytosis and its downstream processes. In vivo, the SF‐MMT nanocomposite hydrogel effectively promotes osteochondral regeneration as evidenced by macroscopic, micro‐CT, and histological evaluation. In conclusion, a functionalized SF‐MMT nanocomposite hydrogel is developed with dual‐lineage bioactivity for osteochondral regeneration, indicating its potential in osteochondral tissue engineering.