1,25-Dihydroxyvitamin D 3 stimulates odontoblastic differentiation of human dental pulp-stem cells in vitro

• Published in: Connective tissue research Vol. 58; no. 6; pp. 531 - 541
• Main Authors: Mucuk, Goksen, Sepet, Elif, Erguven, Mine, Ekmekcı, Ozlem, Bılır, Ayhan
• Format: Journal Article
• Language: English
• Published: England 02.11.2017
• Subjects: Alkaline Phosphatase - metabolism; Calcification, Physiologic - drug effects; Calcitriol - pharmacology; Cell Differentiation - drug effects; Cell Differentiation - physiology; Cell Proliferation - drug effects; Cells, Cultured - drug effects; Dental Pulp - cytology; Humans; Odontoblasts - cytology; Odontoblasts - drug effects; Stem Cells - cytology; Stem Cells - drug effects; dental pulp-stem cells; cell culture; 1,25-Dihydroxyvitamin D3; vitamin D; odontoblastic differentiation
• ISSN: 0300-8207; 1607-8438
• DOI: 10.1080/03008207.2016.1264395

1,25-Dihydroxyvitamin D (1,25-OH D ) plays an important role in mineralized tissue metabolism, including teeth. However, few studies have addressed its role in odontoblastic differentiation of human dental pulp-stem cells (hDPSCs). This study aimed to understand the influence of various concentrations of 1,25-OH D on the proliferation capacity and early dentinogenesis responses of hDPSCs. hDPSCs were obtained from the impacted third molar teeth. Monolayer cultured cells were incubated with a differentiation medium containing different concentrations of 1,25-OH D (0.001, 0.01, and 0.1 µM). All groups were evaluated by S-phase rate [immunohistochemical (IHC) bromodeoxyuridine (BrdU) staining], STRO-1 and dentin sialoprotein (DSP)+ levels (IHC), and alkaline phosphatase (ALP, enzyme-linked immunosorbent assay (ELISA)) levels. The number of cells that entered the S-phase was determined to be the highest and lowest in the control and 0.001 µM 1,25-OH D groups, respectively. The 0.1 µM vitamin D group had the highest increase in DSP+ levels. The highest Stro-1 levels were detected in the control and 0.1 µM 1,25-OH D groups, respectively. The 0.1 µM 1,25-OH D induced a mild increase in ALP activity. This study demonstrated that 1,25-OH D stimulated odontoblastic differentiation of hDPSCs in vitro in a dose-dependent manner. The high DSP + cell number and a mild increase in ALP activity suggest that DPSCs treated with 0.1 μM 1,25-OH D are in the later stage of odontoblastic differentiation. The results confirm that 1,25-OH D -added cocktail medium provides a sufficient microenvironment for the odontoblastic differentiation of hDPSCs in vitro.