Oops they did it again! Carbon nanotubes hoax scientists in viability assays
Oops they did it again! Carbon nanotubes hoax scientists in viability assays
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DOI：
10.1021/nl060177c
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发表时间：
2006-06-01
期刊：
影响因子：
10.8
通讯作者：
Krug, H. F.
中科院分区：
文献类型：
--
作者：
Worle-Knirsch, J. M.;Pulskamp, K.;Krug, H. F.
具有新兴技术重要性的新材料是单壁碳纳米管（SWCNT）。由于单壁碳纳米管将大量用于商业产品，因此其对人类健康和环境的影响已在多项研究中得到解决。已经描述了体内吸入研究和体外浸没应用的不同结果。为什么有些人表明有很强的细胞毒性，而有些人没有，这就是我们在这里报告的原因。来自与碳纳米管孵育的A549细胞的数据在24小时后的MTT测定中伪造了达到大约50%的强细胞毒性效应，而用SWCNT进行相同处理，但用WST-1检测，显示没有细胞毒性。LDH、FACS辅助的线粒体膜电位测定和Annexin-V/PI染色也显示无细胞毒性。单壁碳纳米管似乎与一些四唑盐如MTT相互作用，但不与其他四唑盐（如WST-1、INT、XTT）相互作用。这种干扰似乎不影响酶促反应，而是在于MTT-甲瓒的不溶性。我们的研究结果强烈建议使用至少两个或多个独立的测试系统来验证这类新材料（纳米材料）的细胞毒性数据。此外，我们强烈建议对所用材料进行标准化纳米毒理学测定：显然需要参考材料。MTT反应中形成的MTT-甲瓒晶体与纳米管聚集在一起，并提供了一种潜在的机制来指导生物修复和清除“污染”组织中的单壁碳纳米管。单壁碳纳米管是组织生长的良好支持材料，因为附着的焦点粘连和连接到细胞骨架表明。
New materials of emerging technological importance are single-walled carbon nanotubes (SWCNTs). Because SWCNTs will be used in commercial products in huge amounts, their effects on human health and the environment have been addressed in several studies. Inhalation studies in vivo and submerse applications in vitro have been described with diverging results. Why some indicate a strong cytotoxicity and some do not is what we report on here. Data from A549 cells incubated with carbon nanotubes fake a strong cytotoxic effect within the MTT assay after 24 h that reaches roughly 50%, whereas the same treatment with SWCNTs, but detection with WST-1, reveals no cytotoxicity. LDH, FACS-assisted mitochondrial membrane potential determination, and Annexin-V/PI staining also reveal no cytotocicity. SWCNTs appear to interact with some tetrazolium salts such as MTT but not with others (such as WST-1, INT, XTT). This interference does not seem to affect the enzymatic reaction but lies rather in the insoluble nature of MTT-formazan. Our findings strongly suggest verifying cytotoxicity data with at least two or more independent test systems for this new class of materials (nanomaterials). Moreover, we intensely recommend standardizing nanotoxicological assays with regard to the material used: there is a clear need for reference materials. MTT-formazan crystals formed in the MTT reaction are lumped with nanotubes and offer a potential mechanism to guide bioremediation and clearance for SWCNTs from "contaminated" tissue. SWCNTs are good supporting materials for tissue growth, as attachment of focal adhesions and connections to the cytoskeleton suggest.