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通过单细胞电感耦合等离子体质谱法研究模型纳米塑料的细胞摄取

Investigating the Cellular Uptake of Model Nanoplastics by Single-Cell ICP-MS.

作者信息

Cassano Domenico, Bogni Alessia, La Spina Rita, Gilliland Douglas, Ponti Jessica

机构信息

European Commission, Joint Research Centre (JRC), 21027 Ispra, Italy.

出版信息

Nanomaterials (Basel). 2023 Feb 1;13(3):594. doi: 10.3390/nano13030594.

Abstract

A synthetic route to producing gold-doped environmentally relevant nanoplastics and a method for the rapid and high-throughput qualitative investigation of their cellular interactions have been developed. Polyethylene (PE) and polyvinyl chloride (PVC) nanoparticles, doped with ultrasmall gold nanoparticles, were synthesized via an oil-in-water emulsion technique as models for floating and sedimenting nanoplastics, respectively. Gold nanoparticles were chosen as a dopant as they are considered to be chemically stable, relatively easy to obtain, interference-free for elemental analysis, and suitable for bio-applications. The suitability of the doped particles for quick detection via inductively coupled plasma mass spectrometry (ICP-MS), operating in single-cell mode (scICP-MS), was demonstrated. Specifically, the method was applied to the analysis of nanoplastics in sizes ranging from 50 to 350 nm, taking advantage of the low limit of detection of single-cell ICP-MS for gold nanoparticles. As an initial proof of concept, gold-doped PVC and PE nanoplastics were employed to quantify the interaction and uptake of nanoplastics by the RAW 264.7 mouse macrophage cell line, using scICP-MS and electron microscopy. Macrophages were chosen because their natural biological functions would make them likely to internalize nanoplastics and, thus, would produce samples to verify the test methodology. Finally, the method was applied to assess the uptake by CaCo-2 human intestinal cells, this being a more relevant model for humanexposure to those nanoplastics that are potentially available in the food chain. For both case studies, two concentrations of nanoplastics were employed to simulate both standard environmental conditions and exceptional circumstances, such as pollution hotspot areas.

摘要

已开发出一种合成路线,用于生产掺金且与环境相关的纳米塑料,以及一种对其细胞相互作用进行快速高通量定性研究的方法。通过水包油乳液技术合成了分别掺杂超小金纳米颗粒的聚乙烯(PE)和聚氯乙烯(PVC)纳米颗粒,作为漂浮和沉降纳米塑料的模型。选择金纳米颗粒作为掺杂剂,因为它们被认为化学性质稳定、相对容易获得、对元素分析无干扰且适用于生物应用。证明了掺杂颗粒适用于通过以单细胞模式运行的电感耦合等离子体质谱(ICP-MS)进行快速检测。具体而言,利用单细胞ICP-MS对金纳米颗粒的低检测限,该方法被应用于分析尺寸范围为50至350nm的纳米塑料。作为概念的初步验证,使用掺金的PVC和PE纳米塑料,通过单细胞ICP-MS和电子显微镜来量化RAW 264.7小鼠巨噬细胞系对纳米塑料的相互作用和摄取。选择巨噬细胞是因为它们的天然生物学功能可能使它们易于内化纳米塑料,从而产生样本以验证测试方法。最后,该方法被应用于评估CaCo-2人肠道细胞的摄取情况,这是一个更相关的模型,用于模拟人类接触食物链中可能存在的那些纳米塑料的情况。对于这两个案例研究,采用了两种浓度的纳米塑料来模拟标准环境条件和特殊情况,如污染热点地区。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2709/9920308/65ae4f8200b8/nanomaterials-13-00594-g001.jpg

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