Purpose: To evaluate the effect of microRNA (miR)-124 on osteogenic differentiation of dental pulp mesenchymal stem cells (DPSCs) and to explore the possible mechanism.
Methods: Logarithmic DPSCs were collected and divided into blank group, no-load group, miR-124 inhibitor group, miR-124 inhibitor combined with N-[N-(3,5-difluorophenacetyl)-1-alanyl]-S-ph (DAPT, Notch signaling pathway inhibitor) group. The blank group was not treated, the empty group was transfected with negative control vector inhibitor-NC, the miR-124 inhibitor group was transfected with miR-124 inhibitor, the miR-124 inhibitor combined with DAPT group was transfected with miR-124 inhibitor, and DAPT was added to make the final concentration of 5 μmol/L. The proliferation ability was tested by CCK-8 method 48 h after transfection. Alkaline phosphatase (ALP) activity was tested by p-nitrophenyl phosphate (P-NPP) method after 2 weeks of induction. The area of calcified nodules was tested by alizarin red staining method. The protein expression of hair-like division-related enhancer 1 (HEY1), hair-like division-related enhancer 2 (HEY2), and cyclin D1 gene (CCND1) were tested by Western blot. The data was analyzed by SPSS 19.0 software package.
Results: Compared with the blank group and no-load group, the A450 value at 24, 48, 72 h detected by CCK-8 experiment, A450 value of ALP activity, the area composition ratio of calcified nodules, and expression of HEY1, HEY2, and CCND1 in the miR-124 inhibitor group were increased (P<0.05). Compared with miR-124 inhibitor group, the A450 value at 24, 48, 72 h detected by CCK-8 experiment, A450 value of ALP activity, the area composition ratio of calcified nodules, and the expression of HEY1, HEY2, and CCND1 in the miR-124 inhibitor combined with DAPT group were significantly decreased(P<0.05).
Conclusions: Down-regulation of miR-124 can promote osteogenic differentiation of DPSCs. It is speculated that the mechanism of action is related to the activation of Notch signaling pathway.