ABSTRACT: Canabidiol (CBD), derived from Cannabis sativa plant exhibits beneficial in alleviating a number of of oral and dental pathoses This study aimed to explore the impact of CBD on several traits of human dental pulp stem cell (hDPSC) such as their proliferation, apoptosis, migration, and odonto/osteogenic differentiation. hDPSCs were harvested from human dental pulp tissues. The cells were treated with 1.25, 2.5, 5, 10, 25, and 50 μg/mL CBD. Cell response in terms of cell proliferation, colony-forming unit, cell cycle progression, cell migration, apoptosis, and odonto/osteogenic differentiation of hDPSCs were assessed in normal and P. gingivalis lipopolysaccharide (LPS)-induced `inflammatory` milieus. RNA sequencing and proteomic analysis were performed to predict target pathways impacted by CBD. In a normal physiological growth milieu, exposure to 5 μg/mL CBD attenuated hDPSCs proliferation and reduced the number of colony-forming units. On the contrary, in LPS-exposed hDPSCs, a dose of 1.25 μg/mL CBD restored proliferation and odonto/osteogenic differentiation. Proteomic analysis demonstrated that several proteins involved in cell proliferation and differentiation were upregulated following CBD exposure, including CCL8, CDC42, and KFL5. RNA sequencing data indicated that CBD upregulated Notch signaling pathway. In an inhibitory experiment, DAPT, a Notch inhibitor, reduced the effect of CBD-rescued LPS-attenuated mineralization in hDPSCs, suggesting that CBD potentially mediates Notch activation to exert its effect on odonto/osteogenic differentiation of hDPSCs. CBD recovers the proliferation and survival of hDPSCs following exposure to LPS. Additionally,we report here, for the first time, that CBD-mediated Notch activation effectively restores the odonto/osteogenic differentiation ability of hDPSCs. These results while uncovers and underscore the potential role of CBD as a therapeutic option to enhance dentin regeneration.
