In vitro toxicity evaluation of single walled carbon nanotubes on human A549 lung cells
In vitro toxicity evaluation of single walled carbon nanotubes on human A549 lung cells
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DOI：
10.1016/j.tiv.2006.10.007
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发表时间：
2007-04-01
影响因子：
3.2
通讯作者：
Lyng, Fiona M.
中科院分区：
文献类型：
--
作者：
Davoren, Maria;Herzog, Eva;Lyng, Fiona M.
本文描述了单壁碳纳米管（SWCNT）对人肺细胞系A549细胞的体外细胞毒性评估。采用阿替尼蓝（AB）、中性红（NR）和MTT法测定细胞活力，分别评价代谢、溶酶体和线粒体活性。此外，使用考马斯亮蓝（CB）测定法测量细胞的总蛋白含量。还测定上清液的腺苷酸激酶（AK）释放和白细胞介素8（IL-8），其分别指示细胞膜完整性的丧失和炎症反应。为了研究试验培养基中血清组分与试验材料之间的相互作用，在含血清（5%）和无血清培养基中进行暴露。细胞毒性试验（AB、CB、MTT）的结果显示，SWCNT对A549细胞的急性毒性非常低，因为除一个外，所有报告的24 h EC 50值均超过了试验的最高浓度（800 μ g/ml）。发现单壁碳纳米管干扰细胞毒性评估中使用的许多染料，我们目前正在进行全面的光谱研究，以进一步研究这些相互作用。在使用的多种细胞毒性试验中，发现AB试验最灵敏且可重现。透射电子显微镜（TEM）的研究证实，有没有在A549细胞内的单壁碳纳米管定位后24小时的曝光，但是，在暴露的细胞中观察到的表面活性剂存储板层体的数量增加。(c)2006爱思唯尔有限公司保留所有权利。
This paper describes the in vitro cytotoxicity assessment of single walled carbon nanotubes (SWCNT) on A549 cells, a human lung cell line. Cellular viability was determined using the alamar blue (AB), neutral red (NR) and MTT assays, which evaluated metabolic, lysosomal and mitochondrial activity respectively. In addition, the total protein content of the cells was measured using the coomassie brilliant (CB) blue assay. Supernatants were also assayed for Adenylate Kinase (AK) release and Interleukin 8 (IL-8) which indicated a loss of cell membrane integrity and an inflammation response respectively. To investigate the interactions between serum components in the test medium and the test materials, exposures were conducted both in serum containing (5%) and serum-free medium. Results from the cytotoxicity tests (AB, CB, MTT) revealed the SWCNT to have very low acute toxicity to the A549 cells as all but one of the reported 24 h EC50 values exceeded the top concentration tested (800 mu g/ml). The SWCNT were found to interfere with a number of the dyes used in the cytotoxicity assessment and we are currently conducting a comprehensive spectroscopic study to further investigate these interactions. Of the multiple cytotoxicity assays used, the AB assay was found to be the most sensitive and reproducible. Transmission electron microscopy (TEM) studies confirmed that there was no intracellular localization of SWCNT in A549 cells following 24 h exposure; however, increased numbers of surfactant storing lamellar bodies were observed in exposed cells. (c) 2006 Elsevier Ltd. All rights reserved.