Objective This study aims to explore the effect of bergapten (BP) on the osteogenic differentiation of human dental pulp stem cells (hDPSCs).Methods Specific antigens of hDPSCs were identified by flow cytometry analysis.Cytotoxicity and biosafety of BP was detected by CCK-8 (cell counting kit-8, CCK-8), and 7.5 μM, 15 μM, and 30 μM were selected as the osteogenic differentiation experimental concentrations accordingly. The hDPSCs were randomly divided into control groups and BP groups.Different concentrations of BP were added in complete medium in BP group ,while equal volume of DMSO was added to the complete medium in control group. The morphology of hDPSCs in BP and control groups is detected by fluorescent stain.BCIP/NBT staining and ALP quantitative detection were used to detect the effect of BP at the early stage of osteogenic differentiation of hDPSCs. Alizarin red S (ARS) detection was used to assess the degree of extracellular matrix mineralization at the late stage of .osteogenic differentiation.Expression of osteogenic-related genes such as core binding factor alphal 1(Runx2), osteocalcin (Ocn), osteopontin (Opn), alkaline phosphatase (ALP) were detected by quantitative real-time PCR(qPCR). Expression of osteogenic-related proteins were detected using western blot analysis (Western Blot, WB).Results BP of 7.5 μM, 15 μM and 30 μM showed better biosafety and 15 μM was better (P<0.05).The ALP activity in the 15 μM group was higher than that in control group (P<0.05). The degree of extracellular matrix mineralization of BP group was higher than that in control group and the number of calcium nodules in 15 μM group was more. The expression of osteogenic-related genes and proteins was significantly increased in BP groups and 15 μM group was better (P<0.05). Conclusions BP can promote the osteogenesis differentiation of human dental pulp stem cells in vitro, and the induction effect of 15 μM group was better.