Effect of two different concentrations of 1α,25-dihydroxyvitamin D3 on odontogenic differentiation of stem cells from human exfoliated deciduous teeth

Stem cells from human exfoliated deciduous teeth (SHEDs) can transform into odontoblasts in vitro and in vivo.The role of 1α,25‑dihydroxyvitamin D3 (1α,25 vitD3) has been reported in the mineralization of hard tissues and teeth, as well as osteoblastic differentiation.This study aimed to assess the effect of different concentrations of 1α,25 vitD3 on odontogenic differentiation of SHEDs.

In this experimental study, second‑passage SHEDs were exposed to odontogenic medium along with 0,10,50,100,and 150 nmol concentrations of in 1α,25 vitD3 to determine its optimal concentration for odontogenic differentiation.The methyl thiazolyl tetrazolium (MTT) assay was performed.Odontogenic differentiation was evaluated by QRT‑ polymerase chain reaction for dentin matrix protein (DMP1) and dentin sialophosphoprotein (DSPP) genes. Morphology of differentiated cells was studied by Scanning Electron Microscopy.Data were analyzed using the Kruskal–Wallis,Mann– Whitney, Friedman, and Chi‑square test. P < 0.05 is considered statistically significant.

MTT test result showed the two groups of odontogenic medium + 10 nm 1α,25 vitD3 and odontogenic medium + 150 nm 1α,25 vitD3 provided the most suitable conditions for cell viability at 72 h. Expression of both genes significantly increased in the presence of 1α,25 vitD3 (P < 0.001). Expression of both genes was significantly higher at 14 days compared with 7 days (P < 0.01). At both time points, expression of both genes was significantly higher in the presence of 150 nm 1α,25 vitD3 compared with 10 nm (P < 0.01).The accumulation of cells with odontoblastic morphology, cell interactions, and calcifications were evident.

1α,25 vitD3 upregulates DMP1 and DSPP and results in odontogenic differentiation of SHEDs in odontogenic medium. This upregulation increases with time and by an increase in concentration of 1α,25 vitD3.