牙科复合材料浸出成分对原代人牙龈成纤维细胞的细胞毒性及DNA双链断裂诱导作用
Cytotoxicity and induction of DNA double-strand breaks by components leached from dental composites in primary human gingival fibroblasts.
作者信息
Shehata Mohamed, Durner Jürgen, Eldenez Ayce, Van Landuyt Kirsten, Styllou Panorea, Rothmund Lena, Hickel Reinhard, Scherthan Harry, Geurtsen Werner, Kaina Bernd, Carell Thomas, Reichl Franz X
机构信息
Department of Operative/Restorative Dentistry, Periodontology and Pedodontics, Ludwig-Maximilians-University of Munich, Goethestr. 70, 80336 Munich, Germany.
出版信息
Dent Mater. 2013 Sep;29(9):971-9. doi: 10.1016/j.dental.2013.07.007. Epub 2013 Jul 31.
INTRODUCTION
The public interest steadily increases in the biological adverse effects caused by components released from resin-based dental restorations.
OBJECTIVE
In this study, the cytotoxicity and the genotoxicity were investigated of following released components from dental resin restorations in human gingival fibroblasts (HGF): tetraethyleneglycol dimethacrylate (TEEGDMA), neopentylglycol dimethacrylate (Neopen), diphenyliodoniumchloride (DPIC), triphenyl-stibane (TPSB) and triphenylphosphane (TPP).
METHODS
XTT based cell viability assay was used for cytotoxicity screening of substances. γ-H2AX assay was used for genotoxicity screening. In the γ-H2AX assay, HGFs were exposed to the substances for 6h. Induced foci represent double DNA strand breaks (DSBs), which can induce ATM-dependent phosphorylation of the histone H2AX. Cell death effects (apoptosis and necrosis), induced by the substances were visually tested by the same investigator using the fluorescent microscope.
RESULTS
All tested substances induced a dose-dependent loss of viability in HGFs. Following toxicity ranking among the substances at EC50-concentration were found in the XTT assay (mM, mean±SEM; n=5): DPIC>Neopen>TPSB>TPP>TEEGDMA. DSB-foci per HGF-cell were obtained, when HGFs were exposed to the EC50-concentration of each substance in the following order (mean±SEM; n=3): DPIC>Neopen>TPSB>TPP>TEEGDMA. Multi-foci cells (cells that contain more than 40 foci each) in 80 HGF-cells at EC50-concentration of each substance were found as follow (mean±SEM; n=3): DPIC>Neopen>TPP>TPSB>TEEGDMA. Cell apoptosis contained in each substance at EC50-concentration in the following order (mean±SEM; n=3): DPIC>Neopen>TPSB>TPP >TEEGDMA. Cell necrosis contained in each substance at EC50-concentration in the following order (mean±SEM; n=3): DPIC>Neopen>TPSB>TPP>TEEGDMA.
CONCLUSION
Leached components from dental resin restorations can induce DNA DSBs and cell death effects in HGFs.
引言
公众对树脂基牙科修复体释放成分所引起的生物学不良反应的关注度持续上升。
目的
本研究调查了牙科树脂修复体释放的以下成分在人牙龈成纤维细胞（HGF）中的细胞毒性和遗传毒性：二缩三乙二醇二甲基丙烯酸酯（TEEGDMA）、新戊二醇二甲基丙烯酸酯（Neopen）、二苯基氯化碘鎓（DPIC）、三苯基锑（TPSB）和三苯基膦（TPP）。
方法
采用基于XTT的细胞活力测定法对物质进行细胞毒性筛选。采用γ-H2AX测定法进行遗传毒性筛选。在γ-H2AX测定中，将HGF暴露于这些物质6小时。诱导灶代表双链DNA断裂（DSB），其可诱导组蛋白H2AX的ATM依赖性磷酸化。由同一研究者使用荧光显微镜直观检测这些物质诱导的细胞死亡效应（凋亡和坏死）。
结果
所有测试物质均在HGF中诱导了剂量依赖性的活力丧失。在XTT测定中（mM，平均值±标准误；n = 5），各物质在EC50浓度下的毒性排序如下：DPIC>Neopen>TPSB>TPP>TEEGDMA。当HGF暴露于各物质的EC50浓度时，按以下顺序获得每个HGF细胞中的DSB灶（平均值±标准误；n = 3）：DPIC>Neopen>TPSB>TPP>TEEGDMA。在各物质的EC50浓度下，80个HGF细胞中的多灶细胞（每个细胞含有超过40个灶的细胞）如下（平均值±标准误；n = 3）：DPIC>Neopen>TPP>TPSB>TEEGDMA。各物质在EC50浓度下的细胞凋亡情况按以下顺序（平均值±标准误；n = 3）：DPIC>Neopen>TPSB>TPP>TEEGDMA。各物质在EC50浓度下的细胞坏死情况按以下顺序（平均值±标准误；n = 3）：DPIC>Neopen>TPSB>TPP>TEEGDMA。
结论
牙科树脂修复体渗出的成分可在HGF中诱导DNA双链断裂和细胞死亡效应。
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