Department of Chemistry, The University of Texas at Dallas, Richardson, TX 75080, USA.
J Nanobiotechnology. 2007 Oct 23;5:8. doi: 10.1186/1477-3155-5-8.
This work concerns exposing cultured human epithelial-like HeLa cells to single-walled carbon nanotubes (SWNTs) dispersed in cell culture media supplemented with serum. First, the as-received CoMoCAT SWNT-containing powder was characterized using scanning electron microscopy and thermal gravimetric analyses. Characterizations of the purified dispersions, termed DM-SWNTs, involved atomic force microscopy, inductively coupled plasma - mass spectrometry, and absorption and Raman spectroscopies. Confocal microRaman spectroscopy was used to demonstrate that DM-SWNTs were taken up by HeLa cells in a time- and temperature-dependent fashion. Transmission electron microscopy revealed SWNT-like material in intracellular vacuoles. The morphologies and growth rates of HeLa cells exposed to DM-SWNTs were statistically similar to control cells over the course of 4 d. Finally, flow cytometry was used to show that the fluorescence from MitoSOXtrade mark Red, a selective indicator of superoxide in mitochondria, was statistically similar in both control cells and cells incubated in DM-SWNTs. The combined results indicate that under our sample preparation protocols and assay conditions, CoMoCAT DM-SWNT dispersions are not inherently cytotoxic to HeLa cells. We conclude with recommendations for improving the accuracy and comparability of carbon nanotube (CNT) cytotoxicity reports.
这项工作涉及将培养的人上皮样 HeLa 细胞暴露于在含有血清的细胞培养基中分散的单壁碳纳米管 (SWNTs)。首先，使用扫描电子显微镜和热重分析对所接收的 CoMoCAT 含 SWNT 的粉末进行了表征。对称为 DM-SWNTs 的纯化分散体的表征涉及原子力显微镜、电感耦合等离子体质谱和吸收和拉曼光谱。共焦微拉曼光谱用于证明 DM-SWNTs 以时间和温度依赖的方式被 HeLa 细胞摄取。透射电子显微镜显示 SWNT 样物质在细胞内空泡中。暴露于 DM-SWNTs 的 HeLa 细胞的形态和生长速率在 4 天的过程中与对照细胞在统计学上相似。最后，使用流式细胞术表明，线粒体中超氧阴离子的选择性指示剂 MitoSOXtrade mark Red 的荧光在对照细胞和在 DM-SWNTs 中孵育的细胞中在统计学上相似。综合结果表明，在我们的样品制备方案和测定条件下，CoMoCAT DM-SWNT 分散体对 HeLa 细胞没有固有细胞毒性。我们最后提出了改进碳纳米管 (CNT) 细胞毒性报告的准确性和可比性的建议。
