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一种新型的暴露系统,称为 NAVETTA,用于纳米气溶胶的体外层流电泳沉积,并在人肺报告细胞中评估免疫效应。

A Novel Exposure System Termed NAVETTA for In Vitro Laminar Flow Electrodeposition of Nanoaerosol and Evaluation of Immune Effects in Human Lung Reporter Cells.

机构信息

Flemish Institute for Technological Research (VITO NV) , Boeretang 200, 2400 Mol, Belgium.

Paris Lodron University of Salzburg (PLUS) , Department of Molecular Biology, Hellbrunnerstrasse 34, 5020 Salzburg, Austria.

出版信息

Environ Sci Technol. 2017 May 2;51(9):5259-5269. doi: 10.1021/acs.est.7b00493. Epub 2017 Apr 14.

DOI:10.1021/acs.est.7b00493
PMID:28339192
Abstract

A new prototype air-liquid interface (ALI) exposure system, a flatbed aerosol exposure chamber termed NAVETTA, was developed to investigate deposition of engineered nanoparticles (NPs) on cultured human lung A549 cells directly from the gas phase. This device mimics human lung cell exposure to NPs due to a low horizontal gas flow combined with cells exposed at the ALI. Electrostatic field assistance is applied to improve NP deposition efficiency. As proof-of-principle, cell viability and immune responses after short-term exposure to nanocopper oxide (CuO)-aerosol were determined. We found that, due to the laminar aerosol flow and a specific orientation of inverted transwells, much higher deposition rates were obtained compared to the normal ALI setup. Cellular responses were monitored with postexposure incubation in submerged conditions, revealing CuO dissolution in a concentration-dependent manner. Cytotoxicity was the result of ionic and nonionic Cu fractions. Using the optimized inverted ALI/postincubation procedure, pro-inflammatory immune responses, in terms of interleukin (IL)-8 promoter and nuclear factor kappa B (NFκB) activity, were observed within short time, i.e. One hour exposure to ALI-deposited CuO-NPs and 2.5 h postincubation. NAVETTA is a novel option for mimicking human lung cell exposure to NPs, complementing existing ALI systems.

摘要

一种新型的气液界面(ALI)暴露系统,一种名为 NAVETTA 的平板气溶胶暴露室,被开发出来,以便直接从气相中研究工程纳米颗粒(NPs)在培养的人肺 A549 细胞上的沉积。由于低水平的水平气流与暴露在 ALI 上的细胞相结合,该设备模拟了人类肺细胞对 NPs 的暴露。静电场辅助被应用于提高 NP 沉积效率。作为原理验证,确定了短期暴露于纳米氧化铜(CuO)-气溶胶后细胞活力和免疫反应。我们发现,由于层流气溶胶流和倒置 Transwell 的特定方向,与正常的 ALI 装置相比,获得了更高的沉积速率。用淹没条件下的暴露后孵育来监测细胞反应,揭示了 CuO 以浓度依赖的方式溶解。细胞毒性是离子和非离子 Cu 部分的结果。使用优化的倒置 ALI/后孵育程序,在短时间内观察到促炎免疫反应,即暴露于 ALI 沉积的 CuO-NPs 1 小时和后孵育 2.5 小时。NAVETTA 是模拟人类肺细胞暴露于 NPs 的新选择,补充了现有的 ALI 系统。

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