Effects of tricacium silicate based cements on apical papilla stem cell survive

The purpose of this thesis is to evaluate the effect of tricalcium silicate based cements on apical papilla stem cells (SCAP) in terms of cytotoxicity and odontogenic differentiation. 5x2 discs were obtained from ProRoot WMTA (Dentsply Tulsa Dental Specialties, Johnson City, TN, USA), Neo MTA Plus (Avalon Biomed Inc, Bradenton, FL), Pd MTA White (Produits Dentaires SA, Switzerland) cements containing tricalcium silicate ( n=3). SCAP cells were cultured from wisdom teeth with open apex. The MTS (5-(3- carboxymethoxyphenyl)-2-(4,5-dimethyl thiazole)-3-(4-sulphophenyl) tetrazolium) method was used to evaluate the cytotoxic effect of cements on SCAP. Also, alkaline phosphatase activity was surveyed for odontogenic differentiation. Osteocalcin (OCN), Type I collagen (Col1A1), RUNX2 (Runt-related transcription factor2) gene expressions were analyzed by PCR method. Alizarin Red staining was performed for the visualization of hard tissue formation. The statistical analysis was performed using one-way analysis of variance followed by multiple comparison Anova Tukey test using GraphPad Prism 5 software. In all statistical evaluations, 0.05 was used as the level of significance and accepted that there was a significant difference when test results indicated p<0.05 . Cytotoxicity was tested on the 1st, 4th, 7th and 14th days by the MTS method. No statistically significant difference was observed on the 1st and 4th days. While there was no significant difference at the effect of ProRoot WMTA and Neo MTA Plus groups on viability compared to the control group; a statistically significant decrease was observed in the Pd MTA White group (p<0.05) at 7th and 14th days. Among the MTA varieties, the lowest ALP activity was found to be statistically significant in the Pd MTA White group (p<0.05). Expression of three different genes which were expected to be active in odontogenic differentiation was observed in Neo MTA Plus, Pd MTA White and ProRoot WMTA groups. All MTA varieties were found to be statistically significantly higher than the negative control group (p<0.05). In all MTA groups which were examined in the study, calcium deposits observed with the method Alizarin Red staining. Considering the findings of this in vitro study, it was observed that these three tricalcium silicate-based MTAs had different cytotoxic and odontogenic differentiation effects on SCAP. There is a need for further studies of different tricalcium silicate based cements, both in vivo and in vitro, with various and mutually supportive methods. Keywords: MTA, MTS, Cytotoxicity, Odontogenic differentiation