我们在细胞内 SERS 中实际看到了什么?研究纳米传感器诱导的变化。

What do we actually see in intracellular SERS? Investigating nanosensor-induced variation.

机构信息

Department of Chemistry, Institute of Life Sciences (IfLS), University of Southampton, SO17 1BJ, UK.

出版信息

Faraday Discuss. 2017 Dec 4;205:409-428. doi: 10.1039/c7fd00156h.

Abstract

Plasmonic nanoparticles (NPs), predominantly gold (AuNPs), are easily internalised into cells and commonly employed as nanosensors for reporter-based and reporter-free intracellular SERS applications. While AuNPs are generally considered non-toxic to cells, many biological and toxicity studies report that exposure to NPs induces cell stress through the generation of reactive oxygen species (ROS) and the upregulated transcription of pro-inflammatory genes, which can result in severe genotoxicity and apoptosis. Despite this, the extent to which normal cellular metabolism is affected by AuNP internalisation remains a relative unknown along with the contribution of the uptake itself to the SERS spectra obtained from within so called 'healthy' cells, as indicated by traditional viability tests. This work aims to interrogate the perturbation created by treatment with AuNPs under different conditions and the corresponding effect on the SERS spectra obtained. We characterise the changes induced by varying AuNP concentrations and medium serum compositions using biochemical assays and correlate them to the corresponding intracellular reporter-free SERS spectra. The different serum conditions lead to different extents of nanoparticle internalisation. We observe that changes in SERS spectra are correlated to an increasing amount of internalisation, confirmed qualitatively and quantitatively by confocal imaging and ICP-MS analysis, respectively. We analyse spectra and characterise changes that can be attributed to nanoparticle induced changes. Thus, our study highlights a need for understanding condition-dependent NP-cell interactions and standardisation of nanoparticle treatments in order to establish the validity of intracellular SERS experiments for use in all arising applications.

摘要

等离子体纳米粒子(NPs),主要是金(AuNPs),很容易被细胞内化,并常被用作基于报告分子和无报告分子的细胞内 SERS 应用的纳米传感器。虽然 AuNPs 通常被认为对细胞没有毒性,但许多生物和毒性研究报告称,NP 的暴露会通过产生活性氧(ROS)和上调促炎基因的转录来诱导细胞应激,这可能导致严重的遗传毒性和细胞凋亡。尽管如此,AuNP 内化对正常细胞代谢的影响程度以及摄取本身对所谓“健康”细胞内获得的 SERS 光谱的贡献仍然是一个相对未知的问题,正如传统的生存力测试所表明的那样。这项工作旨在探讨不同条件下用 AuNPs 处理所产生的干扰以及对获得的 SERS 光谱的相应影响。我们使用生化分析方法来描述不同 AuNP 浓度和介质血清成分引起的变化,并将其与相应的无细胞内报告 SERS 光谱相关联。不同的血清条件导致纳米粒子内化的程度不同。我们观察到 SERS 光谱的变化与内化量的增加相关,这通过共聚焦成像和 ICP-MS 分析分别定性和定量地得到证实。我们分析光谱并表征可以归因于纳米粒子诱导变化的变化。因此,我们的研究强调了需要理解条件依赖性 NP-细胞相互作用,并对纳米粒子处理进行标准化,以确立细胞内 SERS 实验在所有出现的应用中的有效性。

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