Abstract

The Portable In Vitro Exposure Cassette (PIVEC) was developed for on-site air quality testing using lung cells. Here, we describe the incorporation of a sensor within the PIVEC for real time monitoring of cellular oxidative stress during exposure to contaminated air. An electrochemical, enzymatic biosensor based on cytochrome c (cyt c) was selected to measure reactive oxygen species (ROS), including hydrogen peroxide and super oxides, due to the stability of signal over time. Human A549 lung cells were grown at the air–liquid interface and exposed within the PIVEC to dry 40 nm copper nanoparticle aerosols for 10 minutes. The generation of ROS compounds was measured during exposure and post-exposure for one hour using the biosensor and compared to intracellular ROS determined using the 2′,7′-dichlorodihydrofluoroscein diacetate (DCFH-DA) assay. A similar increase in oxidative stress upon aerosol exposure was measured using both the cyt c biosensor and DCFH-DA assay. The incorporation of a biosensor within the PIVEC is a unique, first-of-its-kind system designed to monitor the real-time effect of aerosols.

摘要

便携式体外暴露盒 (PIVEC) 是为使用肺细胞进行现场空气质量测试而开发的。在这里，我们描述了在 PIVEC 中加入一个传感器，用于实时监测暴露于受污染空气期间的细胞氧化应激。由于信号随时间的稳定性，选择基于细胞色素 c (cyt c) 的电化学酶生物传感器来测量活性氧 (ROS)，包括过氧化氢和超氧化物。人 A549 肺细胞在气液界面生长，并在 PIVEC 内暴露于干燥的 40 nm 铜纳米颗粒气溶胶中 10 分钟。使用生物传感器在暴露期间和暴露后一小时测量 ROS 化合物的产生，并与使用 2',7'-二氯二氢荧光素二乙酸酯 (DCFH-DA) 测定法测定的细胞内 ROS 进行比较。使用 cyt c 生物传感器和 DCFH-DA 测定法测量了气溶胶暴露后氧化应激的类似增加。在 PIVEC 中加入生物传感器是一种独特的、首创的系统，旨在监测气溶胶的实时影响。