Involvement of AKT/β-catenin in Enhanced Osteogenesis by Cannabidiol in Bone Stromal Cells Harvested from Human Tori.

Cannabidiol (CBD), a major cannabinoid of Cannabis sativa, has been utilized for several medical purposes, such as, an anti-resorptive activity that may benefit treatment of alveolar bone destruction in periodontitis. This study aimed to investigate osteogenic effects of CBD in primary bone stromal cells harvested from jaw tori of healthy patients. To determine cytotoxicity of CBD, the bone stromal cells were treated with various doses of CBD for 24 or 48 h and then analyzed by an alamarBlue(®) assay. No cytotoxicity was found in these cells treated with CBD up to 10 µM. Enhanced osteogenic differentiation and biomineralization of the cells treated with non-toxic doses of CBD were determined by alkaline phosphatase staining and Alizarin Red and von Kossa staining, respectively, and confirmed by upregulated mRNA expressions of runt-related transcription factor 2 (RUNX2), bone sialoprotein (BSP), and Osterix. Treatment with CBD significantly enhanced osteogenic differentiation and biomineralization and upregulated mRNA expressions of RUNX2, BSP, and Osterix (p < 0.05). Pretreatment with MK-2206, an AKT inhibitor, or with CWP232228, a β-catenin antagonist, significantly decreased the increased Alizarin Red staining and the upregulated expressions of BSP and Osterix (p < 0.05), suggesting involvement of AKT/β-catenin in osteogenic induction upon treatment with CBD. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1038/s41598-025-03763-5.