Department of Conservative Dentistry and Endodontics, Manipal College of Dental Sciences, Manipal University, Manipal, Karnataka, India.
Clin Oral Investig. 2013 Jun;17(5):1319-27. doi: 10.1007/s00784-012-0818-x. Epub 2012 Aug 15.
This study aims to evaluate and compare the genotoxic and apoptotic effect of aqueous solutions of ethylenediaminetetraacetic acid (EDTA) with that of maleic acid (MA) using Chinese hamster lung fibroblast (V79) cells growing in vitro.
Exponentially growing V79 cells were treated with various concentrations of EDTA or MA alone for 30 min, and genotoxic effect was analyzed by micronucleus as well as comet assays and the type of cell death by apoptotic cell measurements using microscopic and flow cytometric methods. For all the experiments, H2O2 was used as a positive control.
Treatment of V79 cells with H2O2 resulted in significantly (P < 0.001) increased micronuclei and levels of DNA damage, whereas, EDTA/MA alone treated cells did not show significant increase of MN frequencies and comet parameters even at their higher concentrations when compared with that of untreated control. V79 cells treated with EDTA/MA for 30 min showed a nonsignificant increase in the percentage of apoptotic and necrotic cells at their lower concentrations (0.025 and 0.05 % for EDTA and MA, respectively). However, at higher concentrations, i.e., >IC50 (0.1 and 0.5 %) for EDTA and MA resulted in increased number of apoptotic and necrotic cells when compared with the untreated group.
This study clearly demonstrates that MA and EDTA are not potentially genotoxic agents and MA induced lesser apoptotic/necrotic death than that of EDTA at their clinically relevant doses.
MA may have a better clinical acceptability with comparable smear layer removal ability. Hence, the results presented here might be an additional supporting evidence for the use of MA in endodontic practice.
本研究旨在评估和比较乙二胺四乙酸(EDTA)水溶液与马来酸(MA)的体外遗传毒性和细胞凋亡效应,使用中国仓鼠肺成纤维细胞(V79)。
用不同浓度的 EDTA 或 MA 单独处理指数生长期的 V79 细胞 30 分钟,通过微核分析以及彗星试验分析遗传毒性效应,通过显微镜和流式细胞术分析细胞凋亡的细胞死亡类型。所有实验均使用 H2O2 作为阳性对照。
H2O2 处理 V79 细胞导致微核和 DNA 损伤水平显著增加(P<0.001),而 EDTA/MA 单独处理的细胞即使在较高浓度下也未显示 MN 频率和彗星参数的显著增加,与未处理对照组相比。EDTA/MA 处理 30 分钟的 V79 细胞在较低浓度(分别为 0.025%和 0.05%的 EDTA 和 MA)时,凋亡和坏死细胞的百分比略有增加,但在较高浓度时,即>IC50(0.1%和 0.5%的 EDTA 和 MA)时,与未处理组相比,凋亡和坏死细胞的数量增加。
本研究清楚地表明 MA 和 EDTA 不是潜在的遗传毒性剂,并且在临床相关剂量下 MA 诱导的凋亡/坏死死亡少于 EDTA。
MA 可能具有更好的临床可接受性,并且具有相当的清除玷污层能力。因此,这里提出的结果可能为 MA 在牙髓治疗中的应用提供额外的支持证据。