Characterizing nanoplastics‐induced stress and its SERS fingerprint in an intestinal membrane model
Characterizing nanoplastics‐induced stress and its SERS fingerprint in an intestinal membrane model
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DOI：
10.1002/nano.202100017
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发表时间：
2021-04
期刊：
影响因子：
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通讯作者：
Qianyun Zhang;B. Reinhard
中科院分区：
文献类型：
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作者：
Qianyun Zhang;B. Reinhard
上皮膜提供重要的屏障功能，了解纳米颗粒（NP）暴露如何影响其屏障功能非常重要。在这篇论文中，我们在Caco-2肠上皮膜模型中研究了np诱导的应激及其对细胞外介质振动谱的影响，细胞外介质可以在不干扰细胞的情况下采样和研究。单层cco -2细胞与直径为50 nm的胺或羧基功能化聚苯乙烯（PS） NPs，浓度为1 × 10 12 -1 × 10 14 PS NPs mL -1孵育6和18小时。通过测定活性氧（ROS）生成、细胞活力和肠膜完整性来检测和量化急性暴露条件下PS np诱导的膜损伤。在确定导致np诱导应激的条件后，应用表面增强拉曼光谱（SERS）监测与肠细胞直接接触的培养基的组成，并以微创方式实时检测PS np诱导的细胞代谢的潜在变化。通过人工智能算法和化学计量学工具对SERS光谱进行分析，揭示了PS NPs对细胞代谢的浓度、暴露时间和表面化学依赖性差异。SERS光谱分析确定了次黄嘌呤（c4h4n4o）的环状呼吸模式，作为PS np诱导的膜完整性损失的光谱标记。
Epithelium membranes provide important barrier functions, and it is important to understand how nanoparticle (NP) exposure affects their barrier function. In this manuscript, we investigate NP-induced stress in a Caco-2 intestinal epithelial membrane model and its effect on the vibrational spectrum of the extracellular medium that can be sampled and investigated without perturbation of the cells. Monolayers of Caco-2 cells were incubated with 50 nm diameter polystyrene (PS) NPs functionalized with amine or carboxylic acid groups and concentrations of 1 × 10 12 –1 × 10 14 PS NPs mL –1 for 6 and 18 hours. Reactive oxygen species (ROS) generation, cell viability, and intestinal membrane integrity measurements were performed to detect and quantify PS NP-induced membrane damage under the acute exposure conditions. After identifying conditions that result in NP-induced stress, Surface Enhanced Raman Spectroscopy (SERS) was applied to monitor the composition of the medium in direct contact with the intestinal cells and to detect potential PS NP-induced changes in the cellular metabolism in real time and in a minimally invasive fashion. The analysis of the SERS spectra through artificial intelligence algorithms and chemometric tools revealed concentration, exposure time-, and surface chemistry-dependent differences in the cellular metabolism in response to PS NPs. The SERS spectral analysis identifies the ring breathing mode of hypoxanthine (C 4 H 4 N 4 O), as a spectroscopic marker for the PS NP-induced loss in membrane integrity.