Sonic hedgehog enhances the proliferation and osteogenic differentiation of bone marrow-derived mesenchymal stem cells

Abstract MSCs (mesenchymal stem cells) may be promising seed cells for tissue regeneration because of their self-renewal andmulti-differentiation potential. Shh (sonic hedgehog) is involved in the skeletal formation during embryo developmentand skeletal regeneration. However, how Shh regulates the biological characteristics of BM-MSCs (bone marrow-derivedMSCs) is poorly understood. We have investigated the effect of rShh-N (recombinant N-terminal Shh) on the proliferationand osteogenic differentiation of rBM-MSCs (rat BM-MSCs) in vitro. rBM-MSCs were treated with rShh-N at concentrationsup to 200 ng/ml. Proliferation and colony-forming ability of rBM-MSCs were increased in a dose-dependent manner. rShh-Nincreased the ratio of cells in S and G 2 /M phase, as well as the number of Ki-67 + cells. In addition, ALP (alkalinephosphatase) activity and matrix mineralization were enhanced by 200 ng/ml rShh-N. Real-time PCR showed that rShh-N(200 ng/ml) up-regulated the expression of genes encoding Cbfa-1 (core-binding factor a1), osteocalcin, ALP andcollagen type I in rBM-MSCs. This information reveals some potential of rShh-N in the therapeutics of bone-relateddiseases.