暴露于牙科复合材料浸提液中的人牙龈成纤维细胞的细胞毒性和 DNA 双链断裂。
Cytotoxicity and DNA double-strand breaks in human gingival fibroblasts exposed to eluates of dental composites.
机构信息
Department of Conservative Dentistry and Periodontology, University Hospital, LMU Munich, Germany; Walther-Straub-Institute of Pharmacology and Toxicology, Ludwig-Maximilians-University of Munich, Nußbaumstr. 26, 80336 Munich, Germany.
Department of Orthodontics, Ludwig-Maximilians-University of Munich, Goethestr. 70, 80336 Munich, Germany.
出版信息
Dent Mater. 2018 Feb;34(2):201-208. doi: 10.1016/j.dental.2017.10.002. Epub 2017 Oct 15.
OBJECTIVE
Previously, single composite components were used to study cytotoxicity and induction of DNA double-strand breaks (DNA-DSBs) of dental composite resins. In the present study, cytotoxicity and induction of DNA-DSBs in human gingival fibroblasts (HGFs) were investigated with dental composite eluates consisting of multiple components. The eluates were qualified and quantified.
METHODS
The composites Esthet.X HD, Venus, X-tra fil, CLEARFIL™ AP-X, Admira Fusion and QuiXfil were polymerized and immersed into Dulbecco's modified Eagle's medium (DMEM) for 72h. Subsequently, HGFs were incubated with the corresponding composite eluates. The cell viability of HGFs was obtained from an XTT assay. DNA-DSBs were determined using a γ-H2AX assay. The qualification and quantification of eluates were performed by gas chromatography/mass spectrometry (GC/MS).
RESULTS
HGFs exposed to the eluates of all investigated composites showed no significant loss of cell viability, compared to negative control. Significant DNA-DSBs induction could be found in HGFs exposed to the eluates of Esthet.X HD (0.43±0.05 foci/cell) and Venus (0.39±0.04 foci/cell), compared to control (0.22±0.03 foci/cell). A total of 12 substances were detected from the investigated composite eluates. Five of them were methacrylates: tetraethyleneglycol dimethacrylate (TEGDMA), 2-hydroxyethyl methacrylate (HEMA), hydroxypropyl methacrylate (HPMA), ethyleneglycol dimethacrylate (EGDMA) and trimethylolpropane trimethacrylate (TMPTMA). The highest concentration of HEMA (110.5μM), HPMA (86.08μM) and TMPTMA (4.50μM) was detected in the eluates of QuiXfil. The highest concentration of TEGDMA was 1080μM in Venus eluates and the highest concentration of EGDMA was 3.18μM in Esthet.X HD eluates.
SIGNIFICANCE
Significant DNA-DSBs induction can be found in HGFs exposed to the eluates of Esthet.X HD and Venus. The interactive effects among released (co)monomers and additives may influence the cytotoxicity and induction of DNA-DSBs, compared to exposure with single composite component.
目的
此前，曾使用单一复合成分来研究牙科复合树脂的细胞毒性和诱导 DNA 双链断裂（DNA-DSBs）。本研究采用由多种成分组成的牙科复合浸提液，研究了人牙龈成纤维细胞（HGFs）的细胞毒性和诱导 DNA-DSBs 。对浸提液进行了定性和定量分析。
方法
将 Esthet.X HD、Venus、X-tra fil、CLEARFIL™ AP-X、Admira Fusion 和 QuiXfil 等复合材料聚合后，浸入改良的杜尔贝科氏 Eagle 培养基（DMEM）中 72 小时。随后，用相应的复合浸提液孵育 HGFs。通过 XTT 测定获得 HGFs 的细胞活力。使用 γ-H2AX 测定法测定 DNA-DSBs。通过气相色谱/质谱（GC/MS）对浸提液进行定性和定量分析。
结果
与阴性对照相比，暴露于所有研究用复合浸提液的 HGFs 的细胞活力无明显丧失。与对照组（0.22±0.03 焦点/细胞）相比，暴露于 Esthet.X HD（0.43±0.05 焦点/细胞）和 Venus（0.39±0.04 焦点/细胞）浸提液的 HGFs 中可明显诱导 DNA-DSBs。从研究用复合浸提液中检测到 12 种物质。其中 5 种为甲基丙烯酸酯：三乙二醇二甲基丙烯酸酯（TEGDMA）、2-羟乙基甲基丙烯酸酯（HEMA）、羟丙基甲基丙烯酸酯（HPMA）、乙氧化二甲基丙烯酸酯（EGDMA）和三羟甲基丙烷三甲基丙烯酸酯（TMPTMA）。在 QuiXfil 的浸提液中检测到最高浓度的 HEMA（110.5μM）、HPMA（86.08μM）和 TMPTMA（4.50μM）。Venus 浸提液中 TEGDMA 的最高浓度为 1080μM，Esthet.X HD 浸提液中 EGDMA 的最高浓度为 3.18μM。
意义
暴露于 Esthet.X HD 和 Venus 浸提液的 HGFs 中可明显诱导 DNA-DSBs。与单一复合成分暴露相比，释放（共）单体和添加剂之间的相互作用可能会影响细胞毒性和诱导 DNA-DSBs。
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