Synergistic effects of Indian hedgehog and sonic hedgehog on chondrogenesis during cartilage repair

Sonic hedgehog (Shh) and Indian hedgehog (Ihh) have been shown to control the induction of early cartilaginous differentiation. However, it is unclear whether Ihh and Shh exert synergistic effects on chondrogenesis during articular cartilage repair. Herein, we investigate the effects of chondrogenesis of bone-derived mesenchymal stem cells (BMSCs) following co-transfection with Shh and Ihh via adenoviral vectors in vitro and in vivo. A rotary cell culture system (RCCS) and Cytodex 3 microcarriers were used to create a stereoscopic dynamic environment for cell culture. In the RCCS environment, BMSCs co-transfected with Ihh and Shh displayed stronger chondrogenic differentiation and chondrogenesis than BMSCs transfected with Ihh or Shh alone, and exhibited higher expression levels of Sox 9, ACAN and collagen II, stronger toluidine blue and collagen II immunohistochemical staining. After transplanted into the osteochondral defect at 8 weeks, Ihh/Shh co-transfected BMSCs showed a significantly better cartilage repair than BMSCs transfected with Ihh or Shh alone. Ihh and Shh have synergistic effects on the induction of chondrogenic differentiation and chondrogenesis under a microgravity environment, and help to repair damaged cartilage and reverse subchondral defects during the early stages.