2022, Vol. 26 ›› Issue (30): 4867-4872

Cannabidiol promotes proliferation and osteogenic differentiation of human periodontal ligament stem cells

Li Xiheng1, Li Xinyue1, Mao Tianjiao2, Yang Wanqi3, Tang Liang1, Li Jiang1, 2   

Abstract: BACKGROUND: Cannabidiol, one of the major non-psychotomimetic components from plant Cannabis sativa, has anti-inflammatory, antioxidant, and other pharmacological activities. Recently, it was shown that cannabidiol could effectively promote the osteogenic differentiation of preosteoblasts and mesenchymal stem cells. However, the effect of cannabidiol on the proliferation and osteogenic differentiation of human periodontal ligament stem cells remains unclear.  
OBJECTIVE: To investigate the effects of cannabidiol on the proliferation and osteogenic differentiation of human periodontal ligament stem cells.
METHODS:   Human periodontal ligament stem cells were isolated and cultured using explant method. Cell identification was performed by morphological observation, flow cytometry analysis, and multidirectional differentiation assay. CCK-8 assay was used to assess the effect of different concentrations of cannabidiol (0.1, 0.5, 2.5, 12.5 µmol/L) on the proliferation of human periodontal ligament stem cells. Alkaline phosphatase activity assay, Alizarin Red S staining and semi quantitative analysis were performed to assess different concentrations of cannabidiol (0.1, 0.5, 2.5 µmol/L) on the osteogenic differentiation of human periodontal ligament stem cells. Osteogenic differentiation-related gene expression level changes were assessed by qPCR.  
RESULTS AND CONCLUSION: (1) 0.1 μmol/L cannabidiol showed no significant effect on the viability of human periodontal ligament stem cells; 0.5 and 2.5 μmol/L cannabidiol significantly increased cell proliferation, whereas 12.5 μmol/L cannabidiol had obvious cytotoxicity. (2) 0.1-2.5 μmol/L cannabidiol significantly enhanced alkaline phosphatase activity and mineral deposition of human periodontal ligament stem cells, and upregulated the expression of osteogenic related genes COL 1, OCN, RUNX2, and β-catenin. (3) In conclusion, cannabidiol promoted proliferation and osteogenic differentiation of human periodontal ligament stem cells in a concentration dependent manner, with the most significant effect at 2.5 μmol/L, which may be related to Wnt/β-catenin signaling pathway.
Key words: periodontal ligament stem cells, cannabidiol, osteogenic differentiation, tissue engineering, Wnt/β-catenin signaling pathway