Abstract: Objective To investigate the effect of artesunate (Art) on osteogenic differentiation of dental pulp stem cells (DPSCs) via AMPK/ NF-κB signaling pathway. Methods DPSCs were isolated and identified, the osteogenic and adipogenic differentiation of DPSCs were induced to evaluate the multi-lineage differentiation potential of the cells. DPSCs were treated with different concentrations of Art, and the cell proliferation activity was measured by the CCK-8 method. DPSCs were grouped into 4 groups as follows: the control group, the low-concentration artesunate group (Art-L), the medium-concentration artesunate group (Art-M), and the high-concentration artesunate group (Art-H). Alkaline phosphatase (ALP) staining was performed to measure the ALP activity in cells, and alizarin red S staining were performed to detect the cell mineralization nodules. Western blotting assay was performed to detect the expression levels of osteopontin (OPN), Runt-related transcription factor 2 (RUNX2), osteocalcin (OCN) and AMPK/ NF-κB pathway related proteins. Results In this study, DPSCs were successfully isolated and their multidirectional differentiation potential were verified. The ALP activity, the number of mineralized nodules, the protein expression levels of OPN, RUNX2, OCN, and p-AMPK were significantly increased in DPSCs treated with Art compared with the control group (P< 0. 05), while the expression level of p-NF-κB P65 protein was significantly decreased (P< 0. 05). Conclusion Art can promote the osteogenic differentiation of dental pulp stem cells, which may be related to the regulation of AMPK/ NF-κB signaling pathway. 

Key words: artesunate, dental pulp stem cells, AMPK/ NF-κB signaling pathway, osteogenic differentiation