Department of Pediatric Dentistry and Orthodontics, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Post-Graduation Program in Science and Biotechnology, Universidade Federal Fluminense, Rio de Janeiro, Brazil.
Int Endod J. 2020 Oct;53(10):1348-1373. doi: 10.1111/iej.13353. Epub 2020 Jul 27.
This systematic review aimed to compare the cytotoxicity of root canal filling materials (RCFMs) assessed using tetrazolium salt-based tests (TSBT), including the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, with those obtained using other cell viability assays.
A search was performed on PubMed, Scopus, Web of Science and OpenGrey up to March 2019, followed by a manual search. According to the Participants, Exposure, Comparator and Outcomes (PECO) criteria, in vitro studies that evaluated the cytotoxic effect of RCFMss on animal and/or human cells through TSBT and at least one other viability assay were compared. The methodological quality of selected papers was assessed using ToxRTool and SciRAP . Data were analysed using Wilcoxon's signed-rank test for paired samples and linear weighting kappa.
A total of 230 non-duplicated records were identified. After applying the eligibility criteria, 55 studies were selected for methodological evaluation, seven were selected by manual searching, 22 were excluded for methodological reasons, and 40 were included. A total of 410 comparisons were performed between TSBT and distinct cell viability tests (DCVT). MTT had moderate concordance with DCVT using human cells (n = 138 samples) (P = 0.507; k = 0.4225) and animal cells (n = 122 samples) (P = 0.124; k = 0.5775). XTT had good concordance using human (n = 110 samples) (P = 0.507; k = 0.6336) and animal cells (n = 12 samples) (P = 0.564; k = 0.6604). MTT, XTT, WST and MTS assays showed moderate concordance with DCVT (n = 410 samples) (P = 0.375; k = 0.5138) and complete agreement in 226 samples.
The included studies had methodological heterogeneity that was minimized by the systematic review methodology.
MTT and XTT do not cause over- or underestimation of cell viability during cytotoxicity screening of root canal filling materials, implying that these assays can be considered reliable for this purpose. Nonetheless, the development of protocols for the cytotoxic screening of these materials on 3D tissue-like cultures aiming to improve their predictability in the clinical scenario is suggested.
本系统评价旨在比较基于噻唑蓝盐（TSBT）的测试（包括 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法）与其他细胞活力测定法评估根管充填材料（RCFM）的细胞毒性。
截至 2019 年 3 月，在 PubMed、Scopus、Web of Science 和 OpenGrey 上进行了检索，随后进行了手动检索。根据参与者、暴露、对照和结局（PECO）标准，比较了通过 TSBT 及至少一种其他活力测定法评估 RCFM 对动物和/或人细胞的细胞毒性影响的体外研究。使用 ToxRTool 和 SciRAP 评估所选论文的方法学质量。使用 Wilcoxon 符号秩检验对配对样本和线性加权kappa 进行数据分析。
共确定了 230 份非重复记录。应用入选标准后，对 55 项研究进行了方法学评估，通过手动检索又选择了 7 项研究，因方法学原因排除了 22 项研究，纳入了 40 项研究。在 TSBT 与不同细胞活力测定法（DCVT）之间共进行了 410 次比较。MTT 与人细胞（n=138 个样本）（P=0.507；k=0.4225）和动物细胞（n=122 个样本）（P=0.124；k=0.5775）的 DCVT 具有中度一致性。XTT 与人细胞（n=110 个样本）（P=0.507；k=0.6336）和动物细胞（n=12 个样本）（P=0.564；k=0.6604）的 DCVT 具有良好的一致性。MTT、XTT、WST 和 MTS 测定法与 DCVT（n=410 个样本）（P=0.375；k=0.5138）具有中度一致性，在 226 个样本中具有完全一致性。
纳入的研究存在方法学异质性，通过系统评价方法学将其最小化。
MTT 和 XTT 在对根管充填材料进行细胞毒性筛选时不会导致细胞活力的过高或过低估计，这意味着这些测定法可被认为在这方面是可靠的。然而，建议针对 3D 类组织培养物的这些材料的细胞毒性筛选制定方案，以提高其在临床环境中的预测性。
