Abstract: Objective: To investigate the effect and possible mechanisms of GBE(ginkgo biloba extract)on the differentiation of osteoblast. Methods: Different concentrations of GBE were applied to the MG63. The osteogenic differentiation capacity was evaluated by Alizarin red and alkaline phosphatase (ALP) activity. RT-PCR was taken to test the expression of COL1A1 (type I collagen), OC (osteocalcin), Runx2, and cyclinD1. Western blot was adopted to inspect the protein level of cyclinD1 and subcellular localization of β-catenin. Results: Incubation of MG63 cell in GBE-supplemented culture medium significantly increased ALP activity, the mRNA expression levels of COL1A1, OC, Runx2, and cyclinD1 as well as the cyclinD1 protein level and β-catenin nuclear accumulation at day 7. The matrix mineralization was also augmented after 14 days incubation of GBE. And the 75mg/L was the most effective concentration. Importantly, the positive effects of GBE above were abolished by DKK1, a blocker of the Wnt/β-catenin receptor. Conclusion: GBE has a significantly positive effect on the osteogenic differentiation of the MG63 and the 75mg/L was the most effective concentration, in which Wnt/β-catenin signaling pathway may play an important role.