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二氧化钛纳米颗粒诱导人羊膜上皮(WISH)细胞的细胞毒性、氧化应激和 DNA 损伤。

Titanium dioxide nanoparticles induced cytotoxicity, oxidative stress and DNA damage in human amnion epithelial (WISH) cells.

机构信息

Department of Zoology, College of Science, King Saud University, Riyadh, Saudi Arabia.

出版信息

Toxicol In Vitro. 2012 Mar;26(2):351-61. doi: 10.1016/j.tiv.2011.12.011. Epub 2011 Dec 19.

Abstract

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.

摘要

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。

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