Cannabidiol and Vitamin D3 Impact on Osteogenic Differentiation of Human Dental Mesenchymal Stem Cells

Abstract

Background and objective: The aim of the present study was to establish a new differentiation protocol using cannabidiol (CBD) and vitamin D3 (Vit. D3) for a better and faster osteogenic differentiation of dental tissue derived mesenchymal stem cells (MSCs). Materials and methods: MSCs were harvested from dental follicle (DFSCs), dental pulp (DPSCs), and apical papilla (APSCs) of an impacted third molar of a 17-year old patient. The stem cells were isolated and characterized using flow cytometry; reverse transcription polymerase chain reaction (RT-PCR); and osteogenic, chondrogenic, and adipogenic differentiation. The effects of CBD and Vit. D3 on osteogenic differentiation of dental-derived stem cell were evaluated in terms of viability/metabolic activity by alamar test, expression of collagen1A, osteopontin (OP), osteocalcin (OC), and osteonectin genes and by quantification of calcium deposits by alizarin red assay. Results: Stem cell characterization revealed more typical stemness characteristics for DFSCs and DPSCs and atypical morphology and markers expression for APSCs, a phenotype that was confirmed by differences in multipotential ability. The RT-PCR quantification of bone matrix proteins expression revealed a different behavior for each cell type, APSCs having the best response for CBD. DPSCs showed the best osteogenic potential when treated with Vit. D3. Cultivation of DFSC in standard stem cell conditions induced the highest expression of osteogenic genes, suggesting the spontaneous differentiation capacity of these cells. Regarding mineralization, alizarin red assay indicated that DFSCs and APSCs were the most responsive to low doses of CBD and Vit. D3. DPSCs had the lowest mineralization levels, with a slightly better response to Vit. D3. Conclusions: This study provides evidence that DFSCs, DPSCs, and APSCs respond differently to osteoinduction stimuli and that CBD and Vit. D3 can enhance osteogenic differentiation of these types of cells under certain conditions and doses.

Vitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells

“…A previous report showed that MC3T3-E1 osteoblastic cells showed significant increases in alkaline phosphatase activity with vitamin D at 0.1 nM [34]. Treatment with vitamin D at 5 nM and 10 nM led to a statistically significant increase in Alizarin red optical density of mesenchymal stem cells obtained from dental pulp [35]. Human periodontal ligament stem cells were treated with vitamin D at 100 nM and photobiomodulated, and this led to enhanced osteoblastic differentiation [36].…”

“…The effects of different concentrations of vitamin D have been evaluated in previous studies [34][35][36][37]. A previous report showed that MC3T3-E1 osteoblastic cells showed significant increases in alkaline phosphatase activity with vitamin D at 0.1 nM [34].…”

bFGF stimulated plasminogen activation factors, but inhibited alkaline phosphatase and SPARC in stem cells from apical Papilla: Involvement of MEK/ERK, TAK1 and p38 signaling

“…In this study, bFGF reduced the osteonectin/SPARC production of SCAP. In dental pulp cells, bFGF has been shown to suppress Type I collagen, ALP activity, and SPARC expression [57] . A similar effect of bFGF on murine MC3T3 osteoblasts has also been reported [58] .…”

In quest of a new therapeutic approach in COVID-19: the endocannabinoid system

“… 2019 ; Petrescu et al. 2020 ). The phytocannabinoids are obtained by decarboxylation of the cannabinoid acids within the plants and are represented by cannabigerol (CBG), cannabidivarin (CBGV), cannabidiol (CBD), cannabichromene (CBC), cannabidivarin (CBDV), Δ 9 -tetrahydrocannabivarin (Δ 9 -THCV), and cannabichromevarin (CBCV) (Apostu et al.…”