Real-time monitoring of cellular oxidative stress during aerosol sampling: a proof of concept study

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.

Keywords:

• Biosensor
• copper particles
• inhalation
• nanoparticle
• in vitro

Acknowledgements

The authors acknowledge the Virginia Microelectronics Center for assistance in the fabrication of the biosensor. The author thank Kai Ding for their work in the fabrication process of the sensor including metal deposition and nanostructure growth and Shanzé Eshai and Jessi Shaffer for their work in the functionalization process of the sensor.

Disclosure statement

The authors report no conflict of interest.

Additional information

Funding

This work was supported by startup funds provided to Dr. Lewinski by the College of Engineering at Virginia Commonwealth University. Additionally, LES would like to acknowledge the NIEHS Training Grant [1T32ES019854, PIs: C. Weisel (Contact), G. Mainelis].