School of Dentistry, University of Catanzaro Magna Graecia, Catanzaro, Italy.
Laboratory of Clinical Chemistry and Microbiology, IRCCS Orthopedic Institute Galeazzi, Milano, Italy.
Int Endod J. 2020 Aug;53(8):1093-1110. doi: 10.1111/iej.13314. Epub 2020 May 29.
To evaluate the antimicrobial, toxicity and cleaning effectiveness of ethylenediaminetetraacetic acid (EDTA) and maleic acid (MA) alone and combined with cetrimide (CTR).
Cytotoxic and genotoxic effects were assessed on Chinese hamster cells V79 using the MTT, clonogenic and micronucleus assays, respectively. The bacterial inhibitory and bactericidal concentrations (MIC and MBC, respectively) were determined on a strain of Enterococcus faecalis. Antimicrobial tests were performed on a biofilm model after treatment with the chelating agents by using a biofilm eradication concentration (MBEC) and confocal laser scanning microscope (CLSM) assays. Quantification of cell biomass and percentage of live and dead cells in the biomass were assessed for each group. The percentage reduction of accumulated hard-tissue debris (AHTD) after root canal preparation and final irrigation protocols was evaluated by micro-CT. Statistical tests of one-way analysis of variance (anova), Bonferroni test, Kruskal-Wallis test, Dunn's multiple comparison test and Wilcoxon matched-pairs signed-rank tests were used.
Cetrimide alone as well as in combination with EDTA and MA at dilutions of 1/10 and 1/100 was significantly more toxic as compared to untreated controls (P < 0.001). All tested mixtures were nontoxic at a dilution of 1/1000. EDTA retained a weak inhibitory and bactericidal effect against planktonic cells, whilst MA inhibited cells growth and killed 99.9% of the cells when diluted. CTR revealed the most prominent effect, being inhibitory and bactericidal, also when diluted. Cetrimide alone or combined with EDTA was able to remove, respectively, 40% (P < 0.01) and 60% (P < 0.001) of the entire biomass after 1 min. Conversely, MA alone and in combination with CTR did not have a significant effect on biomass reduction. After final irrigation, the AHTD volume was significantly decreased in all groups (P < 0.05). EDTA + CTR and MA + CTR were associated with a significant reduction in the percentage of AHTD on the entire root canal compared to the same solutions without surfactant.
7% MA was less cytotoxic in comparison with 17% EDTA. The addition of cetrimide to EDTA and MA removed accumulated hard-tissue debris effectively from the canal walls and increased their antimicrobial activity when compared to the same solutions without detergents.
评估乙二胺四乙酸（EDTA）和马来酸（MA）单独使用以及与十六烷基三甲基溴化铵（CTR）联合使用的抗菌、毒性和清洁效果。
使用 MTT、集落形成和微核测定法分别评估中国仓鼠肺细胞 V79 的细胞毒性和遗传毒性效应。在粪肠球菌菌株上测定抑菌浓度（MIC）和杀菌浓度（MBC）。使用生物膜消除浓度（MBEC）和共聚焦激光扫描显微镜（CLSM）测定法，在处理螯合剂后，在生物膜模型上进行抗菌测试。评估每个组的细胞生物量和生物量中活细胞和死细胞的百分比。通过微计算机断层扫描（micro-CT）评估根管预备和最终冲洗方案后硬组织碎屑（AHTD）的累积减少百分比。使用单向方差分析（anova）、Bonferroni 检验、Kruskal-Wallis 检验、Dunn 多重比较检验和 Wilcoxon 配对符号秩检验进行统计检验。
与未处理对照组相比，单独使用十六烷基三甲基溴化铵以及与 EDTA 和 MA 以 1/10 和 1/100 的稀释度混合使用时毒性明显更大（P<0.001）。所有测试的混合物在 1/1000 的稀释度下均无毒。EDTA 对浮游细胞保留弱的抑菌和杀菌作用，而 MA 抑制细胞生长并将 99.9%的细胞杀死，当稀释时。单独的十六烷基三甲基溴化铵或与 EDTA 混合使用时能够分别去除 40%（P<0.01）和 60%（P<0.001）的整个生物量。相反，MA 单独使用和与 CTR 混合使用对生物量减少没有显著影响。最终冲洗后，所有组的 AHTD 体积均显著减少（P<0.05）。与不含表面活性剂的相同溶液相比，EDTA+CTR 和 MA+CTR 与整个根管中 AHTD 百分比的显著减少相关。
与 17%EDTA 相比，7%MA 的细胞毒性更小。与不含清洁剂的相同溶液相比，将十六烷基三甲基溴化铵添加到 EDTA 和 MA 中可以有效地从根管壁上清除累积的硬组织碎屑，并提高其抗菌活性。
