无标记的体外毒性和三种细胞系中柠檬酸稳定的金纳米粒子摄取评估。

Label-free in vitro toxicity and uptake assessment of citrate stabilised gold nanoparticles in three cell lines.

机构信息

Toxicology Section, National Institute for Occupational Health (NIOH), P O Box 4788, Johannesburg 2000, South Africa.

出版信息

Part Fibre Toxicol. 2013 Oct 9;10:50. doi: 10.1186/1743-8977-10-50.

Abstract

BACKGROUND

Reliable in vitro toxicity testing is needed prior to the commencement of in vivo testing necessary for hazard identification and risk assessment of nanoparticles. In this study, the cytotoxicity and uptake of 14 nm and 20 nm citrate stabilised gold nanoparticles (AuNPs) in the bronchial epithelial cell line BEAS-2B, the Chinese hamster ovary cell line CHO, and the human embryonic kidney cell line HEK 293 were investigated.

METHODS

Cytotoxicity of the AuNPs was assessed via traditional XTT-, LDH-, and ATP-based assays, followed by cell impedance studies. Dark-field imaging and hyperspectral imaging were used to confirm the uptake of AuNPs into the cells.

RESULTS

Interference of the AuNPs with the XTT- and ATP-based assays was overcome through the use of cell impedance technology. AuNPs were shown to be relatively non-toxic using this methodology; nevertheless CHO cells were the most sensitive cell type with 20 nm AuNPs having the highest toxicity. Uptake of both 14 nm and 20 nm AuNPs was observed in all cell lines in a time- and cell type-dependent manner.

CONCLUSIONS

Using the cell impedance and dark-field hyperspectral imaging technologies, it was possible to study the toxicity of AuNPs in different cell lines and show that these cells could internalize AuNPs with their subsequent intracellular aggregation. It was also possible to show that this toxicity would not correlate with the level of uptake but it would correlate with cell-type and the size of the AuNPs. Therefore, these two label-free methodologies used in this study are suitable for in vitro studies on the effects of AuNPs, and could present themselves as appropriate and valuable methodologies for future nanoparticle toxicity and uptake studies.

摘要

背景

在进行体内测试以识别和评估纳米颗粒的危害和风险之前,需要进行可靠的体外毒性测试。在这项研究中,研究了 14nm 和 20nm 柠檬酸稳定的金纳米粒子(AuNPs)在支气管上皮细胞系 BEAS-2B、中国仓鼠卵巢细胞系 CHO 和人胚肾细胞系 HEK 293 中的细胞毒性和摄取。

方法

通过传统的 XTT、LDH 和基于 ATP 的测定法评估 AuNPs 的细胞毒性,然后进行细胞阻抗研究。暗场成像和高光谱成像用于确认 AuNPs 进入细胞的摄取。

结果

通过使用细胞阻抗技术克服了 AuNPs 对 XTT 和基于 ATP 的测定的干扰。使用这种方法表明 AuNPs 相对无毒;然而,CHO 细胞是最敏感的细胞类型,20nm AuNPs 的毒性最高。14nm 和 20nm AuNPs 的摄取均在所有细胞系中以时间和细胞类型依赖的方式观察到。

结论

使用细胞阻抗和暗场高光谱成像技术,可以研究不同细胞系中 AuNPs 的毒性,并表明这些细胞可以内化 AuNPs 及其随后的细胞内聚集。还可以表明,这种毒性不会与摄取水平相关,而是与细胞类型和 AuNPs 的大小相关。因此,本研究中使用的这两种无标记方法适用于 AuNPs 对体外研究的影响,并且可以作为未来纳米颗粒毒性和摄取研究的合适和有价值的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4a2d/3853235/54d0354ceebd/1743-8977-10-50-1.jpg

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