Department of Zoology, College of Science, King Saud University, Riyadh, Saudi Arabia.
Toxicol In Vitro. 2012 Mar;26(2):351-61. doi: 10.1016/j.tiv.2011.12.011. Epub 2011 Dec 19.
Titanium dioxide nanoparticles (TiO(2)-NPs) induced cytotoxicity and DNA damage have been investigated using human amnion epithelial (WISH) cells, as an in vitro model for nanotoxicity assessment. Crystalline, polyhedral rutile TiO(2)-NPs were synthesized and characterized using X-ray diffraction (XRD), UV-Visible spectroscopy, Fourier transform infra red (FTIR) spectroscopy, and transmission electron microscopic (TEM) analyses. The neutral red uptake (NRU) and [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) assays revealed the concentration dependent cytotoxic effects of TiO(2)-NPs (30.6nm) in concentration range of 0.625-10μg/ml. Cells exposed to TiO(2)-NPs (10μg/ml) exhibited significant reduction (46.3% and 34.6%; p<0.05) in catalase activity and glutathione (GSH) level, respectively. Treated cells showed 1.87-fold increase in intracellular reactive oxygen species (ROS) generation and 7.3% (p<0.01) increase in G(2)/M cell cycle arrest, as compared to the untreated control. TiO(2)-NPs treated cells also demonstrated the formation of DNA double strand breaks with 14.6-fold (p<0.05) increase in Olive tail moment (OTM) value at 20μg/ml concentration, vis-à-vis untreated control, under neutral comet assay conditions. Thus, the reduction in cell viability, morphological alterations, compromised antioxidant system, intracellular ROS production, and significant DNA damage in TiO(2)-NPs exposed cells signify the potential of these NPs to induce cyto- and genotoxicity in cultured WISH cells.
采用人羊膜上皮(WISH)细胞作为纳米毒性评估的体外模型,研究了二氧化钛纳米粒子(TiO2-NPs)诱导的细胞毒性和 DNA 损伤。采用 X 射线衍射(XRD)、紫外-可见分光光度法、傅里叶变换红外(FTIR)光谱和透射电子显微镜(TEM)分析对结晶多面金红石 TiO2-NPs 进行了合成和表征。中性红摄取(NRU)和[3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴化物](MTT)测定法揭示了 TiO2-NPs(30.6nm)在 0.625-10μg/ml 浓度范围内的浓度依赖性细胞毒性作用。暴露于 TiO2-NPs(10μg/ml)的细胞表现出过氧化氢酶活性和谷胱甘肽(GSH)水平显著降低(分别为 46.3%和 34.6%;p<0.05)。与未处理的对照组相比,处理过的细胞显示出细胞内活性氧(ROS)生成增加 1.87 倍,G2/M 细胞周期阻滞增加 7.3%(p<0.01)。与未处理的对照组相比,中性彗星分析条件下,20μg/ml 浓度下 TiO2-NPs 处理的细胞还表现出 DNA 双链断裂的形成,Olive 尾巴矩(OTM)值增加了 14.6 倍(p<0.05)。因此,细胞活力降低、形态改变、抗氧化系统受损、细胞内 ROS 产生以及 TiO2-NPs 暴露细胞中的显著 DNA 损伤表明这些 NPs 具有在培养的 WISH 细胞中诱导细胞毒性和遗传毒性的潜力。