The effect of accelerated mineral trioxide aggregate on odontoblastic differentiation in dental pulp stem cell niches.

Abstract

To investigate the effect of accelerated-set mineral trioxide aggregate (MTA) on the proliferation and odontoblastic differentiation of human dental pulp cell niches (DPSC). ProRoot White MTA (WMTA; Dentsply Tulsa Dental, Johnson City, TN, USA) was mixed with various additives, which included distilled water, 2.5% disodium hydrogen phosphate (Na2 HPO4 ; Merck, Darmstadt, Germany) and 5% calcium chloride (CaCl2 ; Merck). DPSC niches extracted from third molars were cultured directly on MTA in the culture medium. Cell viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4- sulphophenyl)-2H-tetrazolium (MTS) assay. Cell growth and expression of odontoblastic differentiation markers (dentine sialophosphoprotein (DSPP) and collagen type 1 (COL1)) were determined using Real-Time Polymerase Chain Reaction analysis. Osteo-/odontogenic differentiation of DPSC niches was evaluated by measurement of alkaline phosphatase activity (ALP). Calcium deposition was assessed using von Kossa staining. The results were analysed statistically using Mann-Whitney tests and Kruskal-Wallis tests. MTA mixed with 5% CaCl2 and 2.5% Na2 HPO4 exhibited optimal cell viability (P<0.05) compared to MTA mixed with distilled water. MTA mixed with 5% CaCl2 and 2.5% Na2 HPO4 significantly increased ALP activity (P<0.05), significantly promoted mineralization nodule formation (P<0.05) and significantly enhanced the mRNA expression level of the osteogenic/odontogenic markers (P<0.05; DSPP and COL1) compared with MTA mixed with distilled water. MTA mixed with 5% CaCl2 and 2.5% Na2 HPO4 was biocompatible with dental pulp stem cell niches. Accelerated-set MTA promoted better differentiation in DPSC niches than conventional MTA. The accelerators could provide an alternative to MTA mixed with distilled water.