P Kulan1, O Karabiyik2, G T Kose2,3, B Kargul1. 1. Department of Pediatric Dentistry, Faculty of Dentistry, Marmara University, Istanbul, Turkey. 2. Department of Genetics and Bioengineering, Faculty of Engineering, Yeditepe University, Istanbul, Turkey. 3. BIOMATEN Center of Excellence in Biomaterials and Tissue Engineering, METU, Ankara, Turkey.
Abstract
AIM: To investigate the effect of accelerated-set mineral trioxide aggregate (MTA) on the proliferation and odontoblastic differentiation of human dental pulp cell niches (DPSC). METHODOLOGY: 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. RESULTS: 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. CONCLUSIONS: 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.
AIM: To investigate the effect of accelerated-set mineral trioxide aggregate (MTA) on the proliferation and odontoblastic differentiation of human dental pulp cell niches (DPSC). METHODOLOGY: 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. RESULTS: 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. CONCLUSIONS: 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.
Authors: Shelly Arora; Paul R Cooper; Jithendra T Ratnayake; Lara T Friedlander; Shakila B Rizwan; Benedict Seo; Haizal M Hussaini Journal: Int Endod J Date: 2022-02-04 Impact factor: 5.165