Department of Conservative Dentistry, Federal University of Rio Grande do Sul-UFRGS Faculty of Dentistry, Porto Alegre, Brazil.
Department of Dentistry, Ibirapuera University (UNIB) Faculty of Dentistry, São Paulo, Brazil.
Eur Endod J. 2022 Jun;7(2):129-134. doi: 10.14744/eej.2022.09326.
This study aimed to assess the genotoxicity and cytotoxicity of Sealer Plus BC (SBC), AH Plus (AHP) and MTA Fillapex (MTF).
Human periodontal ligament dental stem cells (hPDLSCs) from third molars were isolated and cultured in a clonogenic medium. Cells were maintained in an incubator, and cell growth was monitored daily. hPDLSCs were characterised under flow cytometry and stem cell surface markers. The tested groups were a control group, SBC, AHP and MTF. Each sealer was prepared according to the manufacturer's instructions and placed in a clonogenic medium to produce a conditioned media. Conditioned media were then diluted to 10% to be placed in contact with culture cells in cell viability assay afterwards. The cells were harvested and plated into 96 wells culture plates. Genotoxicity was assessed by evaluation of micronucleus formation and cytotoxicity by MTT-based assay. All experiments were performed in triplicate. Data normality was verified by the Kolmogorov-Smirnov test. Statistical analysis for genotoxicity was performed with Kruskal-Wallis and Dunn's tests and two-way ANOVA for cytotoxicity, both with a significance level of 5%.
Cells expressed typical levels of mesenchymal stem cell surface markers. No differences in the number of micronuclei were observed among all groups (P>0.05). In all periods analysed (24, 48, and 72 h), the sealers presented statistically different results for cell viability (P<0.05), with SBC presenting the lowest cytotoxicity, followed by the control group, MTF, and AHP.
All sealers presented low genotoxicity, and Sealer Plus BC presented the lowest cytotoxicity.
本研究旨在评估 Sealer Plus BC（SBC）、AH Plus（AHP）和 MTA Fillapex（MTF）的遗传毒性和细胞毒性。
从第三磨牙中分离和培养人牙周膜牙髓干细胞（hPDLSCs）于集落形成培养基中。将细胞置于孵育箱中，每天监测细胞生长情况。通过流式细胞术和干细胞表面标志物对 hPDLSCs 进行鉴定。实验组为对照组、SBC、AHP 和 MTF。根据制造商的说明制备每种密封剂，并将其放置在集落形成培养基中以产生条件培养基。然后将条件培养基稀释至 10%，与细胞活力测定中的培养细胞接触。收集细胞并接种于 96 孔培养板中。通过评估微核形成来评估遗传毒性，通过 MTT 法评估细胞毒性。所有实验均重复 3 次。通过 Kolmogorov-Smirnov 检验验证数据正态性。采用 Kruskal-Wallis 和 Dunn 检验进行遗传毒性的统计分析，采用双向方差分析进行细胞毒性的统计分析，均具有 5%的显著性水平。
细胞表达典型的间充质干细胞表面标志物。所有组之间的微核数量均无差异（P>0.05）。在所有分析的时间段（24、48 和 72 h），密封剂的细胞活力均呈现出统计学差异（P<0.05），其中 SBC 的细胞毒性最低，其次是对照组、MTF 和 AHP。
所有密封剂的遗传毒性均较低，而 Sealer Plus BC 的细胞毒性最低。
