Odonto/Osteogenic Differentiation of Dental Pulp Stem Cells of Type 1 Diabetic Patients with Mineral Trioxide Aggregate/1α,25-Dihydroxyvitamin D3 Combination.

INTRODUCTION

Vitamin D3 plays an important role in the mineralization mechanism and is often deficient in diabetic patients. The objective of this study was to investigate the odonto/osteogenic differentiation potential of the combination of mineral trioxide aggregate (MTA)/1α,25-dihydroxyvitamin D3 (VD3) on dental pulp stem cells (DPSCs) of patients with type 1 diabetes mellitus (T1DM).

METHODS

DPSCs isolated from donors (control and T1DM) were cultured and characterized. Cell proliferation and wound healing assays were performed. DPSCs were exposed to 4 different media: growth medium (Dulbecco's modified Eagle's medium, 10% fetal bovine serum, antibiotic, and antimycotic), differentiation medium (DM) (growth medium plus ß-glycerophosphate and ascorbic acid), DM + MTA (DM plus 0.02 mg/mL MTA), and DM + MTA + VD3 (DM + MTA and 10 nmol/L vitamin D3). Odonto/osteogenic differentiation of DPSCs was evaluated by the alizarin red test, relative real-time polymerase chain reaction (dentin sialophosphoprotein, dentin matrix protein 1, collagen type 1 alpha 1, and osteocalcin), immunocytochemistry (antibone sialoprotein II, anti-dentin matrix protein 1, and anti-collagen type 1 alpha 1), and Western blot (dentin matrix protein 1 and osteocalcin) methods.

RESULTS

The proliferation rates of DPSCs isolated from controls were significantly higher than DPSCs isolated from T1DM in a time-dependent manner (P < .05). Alizarin red staining and the expression of odonto/osteogenic markers showed that odonto/osteogenic differentiation was more pronounced in controls (P < .05) compared with T1DM patients. Although DM + MTA caused the odonto/osteogenic differentiation in DPSCs derived from controls, DM + MTA + VD3 resulted in the odonto/osteogenic differentiation in DPSCs of T1DM patients (P < .05).

CONCLUSIONS

Odonto/osteogenic differentiation was affected by both supplements used for differentiation and the systemic disease, diabetes mellitus. The differentiation potential of T1DM-derived DPSCs was clearly increased with the VD3 supplement, although it was not as efficient as in the controls. The VD3 supplement showed a positive effect on the differentiation of T1DM DPSCs compared with MTA alone.