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铜锌合金纳米颗粒对人肺上皮细胞的体外细胞毒性和遗传毒性评估。

Evaluation of in vitro cytotoxicity and genotoxicity of copper-zinc alloy nanoparticles in human lung epithelial cells.

作者信息

Kumbıçak Umit, Cavaş Tolga, Cinkılıç Nilüfer, Kumbıçak Zübeyde, Vatan Ozgür, Yılmaz Dilek

机构信息

Department of Molecular Biology and Genetics, Faculty of Science and Art, Nevşehir University, 50300 Nevşehir, Turkey.

Cell Culture and Genetic Toxicology Laboratory, Department of Biology, Faculty of Sciences and Arts, Uludağ University, 16059 Nilüfer, Bursa, Turkey.

出版信息

Food Chem Toxicol. 2014 Nov;73:105-12. doi: 10.1016/j.fct.2014.07.040. Epub 2014 Aug 10.

Abstract

In the present study, in vitro cytotoxic and genotoxic effect of copper-zinc alloy nanoparticles (Cu-Zn ANPs) on human lung epithelial cells (BEAS-2B) were investigated. XTT test and clonogenic assay were used to determine cytotoxic effects. Cell death mode and intracellular reactive oxygen species formations were analyzed using M30, M65 and ROS Elisa assays. Genotoxic effects were evaluated using micronucleus, comet and γ-H2AX foci assays. Cu-Zn ANPs were characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS) and zeta potential measurements. Characterization of Cu-Zn ANPs showed an average size of 200nm and zeta potential of -22mV. TEM analyses further revealed the intracellular localization of Cu-Zn ANPs in cytoplasm within 24h. Analysis of micronucleus, comet and γ-H2AX foci counts showed that exposure to Cu-Zn ANPs significantly induced chromosomal damage as well as single and double stranded DNA damage in BEAS-2B cells. Our results further indicated that exposure to Cu-Zn ANPs significantly induced intracellular ROS formation. Evaluation of M30:M65 ratios suggested that cell death was predominantly due to necrosis.

摘要

在本研究中,研究了铜锌合金纳米颗粒(Cu-Zn ANPs)对人肺上皮细胞(BEAS-2B)的体外细胞毒性和遗传毒性作用。采用XTT试验和克隆形成试验来确定细胞毒性作用。使用M30、M65和ROS酶联免疫吸附测定法分析细胞死亡模式和细胞内活性氧的形成。使用微核、彗星试验和γ-H2AX焦点试验评估遗传毒性作用。通过透射电子显微镜(TEM)、动态光散射(DLS)和zeta电位测量对Cu-Zn ANPs进行表征。Cu-Zn ANPs的表征显示平均粒径为200nm,zeta电位为-22mV。TEM分析进一步揭示了24小时内Cu-Zn ANPs在细胞质中的细胞内定位。微核、彗星试验和γ-H2AX焦点计数分析表明,暴露于Cu-Zn ANPs会显著诱导BEAS-2B细胞中的染色体损伤以及单链和双链DNA损伤。我们的结果进一步表明,暴露于Cu-Zn ANPs会显著诱导细胞内活性氧的形成。M30:M65比值评估表明,细胞死亡主要是由于坏死。

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