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Graphene oxide nanoparticle attachment and its toxicity on living lung epithelial cells

机译：氧化石墨烯纳米颗粒附着及其对活肺上皮细胞的毒性

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Since its discovery, graphene and its oxidized form, graphene oxide (GO), have attracted interest in a wide range of technical applications. Concerns about their potential toxicity calls for scrutinized studies, but hitherto conflicting results have been reported which partly may be due to variations of synthesis and exposure procedures. Here we report on the attachment and toxicity of contamination-free graphene oxide nanoparticles (GONP) in living lung epithelial cells. The synthesis of chemically pure GONP was made by an improvement of the Hummer's method based on graphene exfoliated from graphite using high-intensity ultrasonication, resulting in two dimensional sheets with a lateral dimension in the range 200 nm to 3 mu m and thickness of 0.9 nm. Confocal Raman spectroscopy combined with multivariate analysis was used to study the interaction of GONP and living cells. It is shown that overlapping Raman bands due to GONPs and biomolecules in the cells can clearly be separated with this approach. Orthogonal partial least squares discriminant analysis was used to compare spectral data collected from cells exposed to GONP with spectral data collected from non-exposed control cells, and spectral data from cells exposed to a surfactant known to induce apoptosis. Our analyses show that GONP readily attach to the cells, forming sheets which cover a large fraction of the cell surfaces, and induce small chemical changes. In particular, chemical modifications of proteins and lipids in lung epithelial cells are inferred. GONPs do not, however, decrease cell viability. In contrast, enhanced cell proliferation is observed. Our results shed new light on the interactions of GO, and in contrast to some previous reports, suggest that GO is not toxic. The hyperspectral Raman spectroscopy analysis employed here should be applicable for other fields in nanomedicine as a label-free non-perturbing analytical method.
机译：自发现以来，石墨烯及其氧化形式的氧化石墨烯（GO）在广泛的技术应用中引起了人们的兴趣。有关其潜在毒性的担忧需要进行仔细研究，但迄今已有报道相互矛盾的结果，部分原因可能是合成方法和暴露程序的差异。在这里，我们报告生活肺上皮细胞中的无污染氧化石墨烯纳米颗粒（GONP）的附着和毒性。化学纯GONP的合成是通过改良Hummer方法进行的，该方法基于高强度超声处理，从石墨剥离的石墨烯上剥离了石墨烯，得到了二维片材，其横向尺寸在200 nm至3μm范围内，厚度为0.9 nm 。共焦拉曼光谱结合多变量分析用于研究GONP与活细胞的相互作用。结果表明，通过这种方法可以清楚地分离出细胞中由于GONP和生物分子而产生的重叠拉曼谱带。正交偏最小二乘判别分析用于比较从暴露于GONP的细胞收集的光谱数据与从未暴露的对照细胞收集的光谱数据，以及暴露于已知可诱导凋亡的表面活性剂的细胞的光谱数据。我们的分析表明，GONP易于附着在细胞上，形成覆盖大部分细胞表面的薄片，并引起小的化学变化。特别地，推断肺上皮细胞中蛋白质和脂质的化学修饰。但是，GONP不会降低细胞活力。相反，观察到细胞增殖增强。我们的研究结果为GO的相互作用提供了新的思路，并且与以前的一些报道形成对比，表明GO无毒。此处采用的高光谱拉曼光谱分析应作为无标记，无干扰的分析方法，可应用于纳米医学的其他领域。

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