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Nanotoxicity of Silver Nanoparticles on HEK293T Cells: A Combined StudyUsing Biomechanical and Biological Techniques

机译:银纳米颗粒对HEK293T细胞的纳米毒性:联合研究使用生物力学和生物技术

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摘要

Human embryonic kidney 293T cells (HEK293T cells) before and after treatment with silver nanoparticles (AgNPs) were measured using advanced atomic force microscopy (AFM) force measurement technique, and the biomechanical property of cells was analyzed using a theoretical model. The biomechanical results showed that the factor of viscosity of untreated HEK293T cells reduced from 0.65 to 0.40 for cells exposure to 40 μg/mL of AgNPs. Comet assay indicated that significant DNA damage occurred in the treated cells, measured as tail DNA% and tail moment. Furthermore, gene expression analysis showed that for the cells treated with 40 μg/mL of AgNPs, the antiapoptosis genes Bcl2-t and Bclw were, respectively, downregulated to 0.65- and 0.66-fold of control, and that the proapoptosis gene Bid was upregulated to 1.55-fold of control, which indicates that apoptosis occurred in cells exposed to AgNPs. Interestingly, excellent negative correlations were found between the factor of viscosity and tail DNA%, and tail moment, which suggest that the biomechanical property can be correlated with genotoxicity of nanoparticles on the cells. Based on the above results, we conclude that (1) AgNPs can lead to biomechanicalchanges in HEK293T cells, concomitantly with biological changes includingcell viability, DNA damage, and cell apoptosis; (2) the factor ofviscosity can be exploited as a promising label-free biomechanicalmarker to assess the nanotoxicity of nanoparticles on the cells; and(3) the combination of AFM-based mechanical technique with conventionalbiological methods can provide more comprehensive understanding ofthe nanotoxicity of nanoparticles than merely by using the biologicaltechniques.
机译:使用先进的原子力显微镜(AFM)力测量技术测量了用银纳米颗粒(AgNPs)处理之前和之后的人类胚胎肾293T细胞(HEK293T细胞),并使用理论模型分析了细胞的生物力学特性。生物力学结果表明,对于暴露于40μg/ mL AgNPs的细胞,未经处理的HEK293T细胞的粘度因子从0.65降低至0.40。彗星试验表明,经处理的细胞发生了明显的DNA损伤,以尾部DNA%和尾部矩来衡量。此外,基因表达分析表明,对于用40μg/ mL AgNPs处理的细胞,抗凋亡基因Bcl2-t和Bclw分别下调至对照的0.65和0.66倍,而促凋亡基因Bid上调达到对照的1.55倍,这表明凋亡发生在暴露于AgNPs的细胞中。有趣的是,在粘度和尾巴DNA%以及尾矩之间发现了极好的负相关性,这表明生物力学特性可以与纳米粒子对细胞的遗传毒性相关。根据以上结果,我们得出结论:(1)AgNPs可以导致生物力学HEK293T细胞发生变化,并伴随生物学变化,包括细胞活力,DNA损伤和细胞凋亡; (2)因素粘度可以用作有前途的无标记生物力学评估纳米颗粒对细胞的纳米毒性的标记物;和(3)基于AFM的机械技术与常规技术的结合生物学方法可以提供更全面的了解纳米颗粒的纳米毒性比仅仅通过生物技术。

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