Department of Histology, National University of Cordoba, Cordoba, Argentina.
J Adhes Dent. 2011 Aug;13(4):375-81. doi: 10.3290/j.jad.a20141.
To analyze the cytotoxic effects of 2-hydroxyethylmethacrylate (HEMA) in human gingival fibroblasts using quantitative x-ray microanalysis (EXPMA) and two classical methods (DNA and LDH release in culture medium).
Different concentrations of HEMA (5, 10, 20, 30, and 40 mM) in DMEM medium were used and the effects on human gingival fibroblasts after 6, 12, and 24 h were determined. As controls, fibroblasts cultured with DMEM culture medium (negative control) and fibroblast incubated in 1% triton X (positive control) were used.
The results showed that correlation between the concentrations of HEMA and the amount of LDH and DNA released to the medium were statistically significant for all times analyzed. LDH and DNA released from cells incubated in the lowest concentrations of HEMA (5 and 10 mM) were not significantly different to negative controls. In contrast, cells incubated in the highest HEMA concentrations (20, 30, 40 mM) showed a significant increase of both LDH and DNA released to the culture medium at 6, 12, and 24 h. On the other hand, the ionic concentration of the different elements analyzed in this work revealed that the contents of P, S, Cl, and K were significantly higher in the controls than in samples incubated for 6 h in 5 mM or 10 mM HEMA (p < 0.01). K/ Na index (an excellent marker of cell viability) showed a significant decrease, and therefore, viability was significantly reduced.
The results suggest that EXPMA is a sensitive method that is able to detect early cell damage even before the cell membrane is altered.
使用定量 X 射线微分析（EXPMA）和两种经典方法（培养基中 DNA 和 LDH 的释放）分析 2-羟乙基甲基丙烯酸酯（HEMA）对人牙龈成纤维细胞的细胞毒性作用。
使用不同浓度的 HEMA（5、10、20、30 和 40 mM）在 DMEM 培养基中，并确定其对人牙龈成纤维细胞在 6、12 和 24 h 后的影响。作为对照，使用在 DMEM 培养基中培养的成纤维细胞（阴性对照）和在 1% Triton X 孵育的成纤维细胞（阳性对照）。
结果表明，HEMA 浓度与培养基中 LDH 和 DNA 释放量之间的相关性在所有分析时间均具有统计学意义。在最低浓度的 HEMA（5 和 10 mM）孵育的细胞中释放的 LDH 和 DNA 与阴性对照无显著差异。相比之下，在最高 HEMA 浓度（20、30、40 mM）孵育的细胞在 6、12 和 24 h 时均显示出培养基中 LDH 和 DNA 释放量的显著增加。另一方面，对本工作中分析的不同元素的离子浓度进行分析表明，在 5 mM 或 10 mM HEMA 孵育 6 h 的样品中，P、S、Cl 和 K 的含量明显高于对照（p < 0.01）。K/Na 指数（细胞活力的一个极好标志物）明显下降，因此，细胞活力明显降低。
结果表明，EXPMA 是一种敏感的方法，即使在细胞膜改变之前，也能够检测到早期的细胞损伤。
