Dental composite components induce DNA-damage and altered nuclear morphology in gingiva fibroblasts.
Dental composite components induce DNA-damage and altered nuclear morphology in gingiva fibroblasts.
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DOI：
10.1016/j.dental.2015.08.156
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发表时间：
2015-11
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影响因子：
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通讯作者：
Marianthi Styllou;F. Reichl;Panorea Styllou;Ebru Urcan;Lena Rothmund;R. Hickel;C. Högg;H. Scherthan
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文献类型：
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作者：
Marianthi Styllou;F. Reichl;Panorea Styllou;Ebru Urcan;Lena Rothmund;R. Hickel;C. Högg;H. Scherthan
释放的牙科复合材料组分可损伤人牙龈成纤维细胞（HGFs）及其DNA。采用改进的γ-H2 AX焦点试验研究了牙科复合材料不同化合物的细胞毒性、染色质凝聚和诱导DNA双链断裂（DSB）。方法HGFs与单体一起孵育：双酚-A-乙氧基化物-二甲基丙烯酸酯（Bis-DMA），双酚-A-甘油酸酯-二甲基丙烯酸酯（BisGMA），乙基三甘醇甲基丙烯酸酯（ETEGMA），甲基丙烯酸缩水甘油酯（GMA），1，6-己二醇-二甲基丙烯酸酯（HDDMA），丙烷乙氧基化物三丙烯酸酯（TMPTA）和丙烯酰胺（ACR）。结果DSB的诱导作用呈浓度依赖性，其顺序为：GMA > BisGMA > ACR > Bis-DMA > HDDMA > TMPTA > ETEGMA。HGF暴露于GMA（0.3 mM）和BisGMA（0.09 mM）诱导最高速率的DSB病灶，即相对于对照（0.33 DSB病灶/细胞）分别为12倍和8倍。在最高浓度（EC 50）下，观察到HGF细胞核染色质形态的显著变化，即核染色质致密化和卵圆形成纤维细胞核覆盖面积减少。GMA（0.3 mM时为1.7倍）和BisGMA（0.09 mM时为1.6倍）可显著诱导细胞核浓缩，这与诱导的DSB灶的最高数量相关（GMA、BisGMA分别为3.9和2.6个灶/细胞）。此外，观察到HGF细胞核形态的浓度依赖性变化，证明了供试物质的遗传毒性作用。
ObjectiveReleased dental composite components can damage human gingival fibroblasts (HGFs) and their DNA. The cytotoxicity, chromatin condensation and the induction of DNA double strand breaks (DSBs) by different compounds of dental composites was investigated using an improved γ-H2AX focus assay.MethodsHGFs were incubated with the monomers: bisphenol-A-ethoxylate-dimethacrylate (Bis-DMA), bisphenol-A-glycerolate-dimethacrylate (BisGMA), ethyltriethylen glycol methacrylate (ETEGMA), glycidyl methacrylate (GMA), 1,6-hexandiol-dimethycrylate (HDDMA), trimethylolpropane ethoxylate triacrylate (TMPTA), and acrylamide (ACR). DSBs were determined by enumerating γ-H2AX and 53BP1 foci colocalized at DSBs.ResultsA concentration-dependent induction of DSBs was found in the order: GMA > BisGMA > ACR > Bis-DMA > HDDMA > TMPTA > ETEGMA. HGFs exposure to GMA (0.3 mM) and to BisGMA (0.09 mM) induced the highest rate of DSB foci, i.e. 12-fold and 8-fold, respectively, relative to control (0.33 DSB foci/cell). At the highest concentrations (EC50) prominent changes in the chromatin morphology of HGF cell nuclei, i.e. compaction of nuclear chromatin and reduction of the area covered by the ovoid fibroblast nuclei, were observed. Nuclear condensation was significantly induced by GMA (1.7-fold at 0.3 mM) and BisGMA (1.6-fold at 0.09 mM), which correlated with the highest numbers of induced DSB foci (GMA, BisGMA, 3.9 and 2.6 foci/cell, respectively).SignificanceThe improved γ-H2AX/53BP1 focus assay revealed a concentration-dependent increase in DSBs for all tested substances. Furthermore, concentration-dependent changes in HGF cell nucleus morphology was noted, demonstrating genotoxic effects of the substances tested.