Enzymatically crosslinked silk-nanosilicate reinforced hydrogel with dual-lineage bioactivity for osteochondral tissue engineering

Osteochondral defects are characterized by damage to both articular cartilage and subchondral bone. Various tissue engineering strategies have been developed for osteochondral defect repair. However, strong mechanical properties and dual-lineage (osteogenesis and chondrogenesis) bioactivity still pose challenges for current biomaterial design. Silicate nanoclay has been reported to improve the mechanical properties and biofunctionality of polymer systems, but its effect on in vitro dual-lineage differentiation or in vivo osteochondral regeneration has not been extensively investigated before. Here, a novel enzymatically crosslinked silk fibroin (SF)-Laponite (LAP) nanocomposite hydrogel was fabricated and evaluated for osteochondral regeneration. The incorporation of a small amount of LAP (1% w/v) accelerated the gelation process of SF and greatly enhanced the mechanical properties and hydrophilicity of the hydrogel. In vitro investigations showed that the developed SF-LAP hydrogel was biocompatible and was able to induce osteogenic and chondrogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs), validated by Alizarin red/Alcian blue staining, qPCR, and immunofluorescent staining. During an 8-week implantation into rabbit full-thickness osteochondral defects, the SF-LAP hydrogel promoted the simultaneous and enhanced regeneration of cartilage and subchondral bone. The repaired tissue in the chondral region was constituted mainly of hyaline cartilage with typical chondrocyte morphology and cartilaginous extracellular matrix (ECM). These findings suggested that the SF-LAP nanocomposite hydrogel developed in this study served as a promising biomaterial for osteochondral regeneration due to its mechanical reinforcement and dual-lineage bioactivity.