Abstract: Objective To analysis the effect of Shh on BMP9-induced osteogenic differentiation of C3H10T1/2 mesenchymal stem cells and the mechanism involved. Methods C3H10T1/2 cells were treated with adenovirus Shh or/and BMP9, then the ALP activity was detected by quantitative and staining assay, calcium deposition was detected by Alizarin Red S staining, the expressions of Shh, BMP9, OPN, OCN, Id1, Id2, Id3, CTGF and Runx2 were detected by RT-PCR, the expressions of BMP9, OPN, OCN, Runx2, Dlx5 and p-Smad1/5/8 were detected by Western blot, Luciferae reporter assay was detected by quantitative. Results Shh can not influence the expression of BMP9 but increase the ealy and later osteogenic differentiation and the expression of osteogenesis related genes of C3H10T1/2 cells induced by BMP9. Furthermore, Shh also led to luciferase activity but not the phosphorylation activity of canonical Smad pathway stimulated by BMP9. Conclusion Shh promotes BMP9-induced osteogenic differentiation of C3H10T1/2 mesenchymal stem cells.

Key words: Key words: mesenchymal stem cells, BMP9, Shh, osteogenic differentiation