In Vitro primary human airway epithelial whole exhaust exposure

Katherine R Landwehr; Jessica Hillas; Ryan Mead-Hunter; Peter Brooks; Andrew King; Rebecca A O'Leary; Anthony Kicic; Benjamin J Mullins; Alexander N Larcombe; AusREC; WAERP

doi:10.1016/j.mex.2021.101561

In Vitro primary human airway epithelial whole exhaust exposure

MethodsX. 2021 Oct 26:8:101561. doi: 10.1016/j.mex.2021.101561. eCollection 2021.

Authors

Katherine R Landwehr^{1

2}, Jessica Hillas², Ryan Mead-Hunter¹, Peter Brooks³, Andrew King⁴, Rebecca A O'Leary⁵, Anthony Kicic^{1

2

6

7}, Benjamin J Mullins¹, Alexander N Larcombe^{1

2}; AusREC; WAERP

Affiliations

¹ Occupation, Environment and Safety, School of Population Health, Curtin University, PO Box U1987, Perth, Western Australia 6845, Australia.
² Respiratory Environmental Health, Wal-yan Respiratory Research Centre, Telethon Kids Institute, Perth Children's Hospital, Nedlands, Perth, Western Australia 6009, Australia.
³ School of Science, Technology and Engineering, University of the Sunshine Coast, Sippy Downs, Queensland, Australia.
⁴ Fluid Dynamics Research Group, School of Civil and Mechanical Engineering, Curtin University, Perth, Western Australia, Australia.
⁵ Department of Primary Industries and Regional Development, Perth, Western Australia 6151, Australia.
⁶ Department of Respiratory and Sleep Medicine, Perth Children's Hospital, Nedlands, Perth, Western Australia 6009, Australia.
⁷ Centre for Cell Therapy and Regenerative Medicine, The University of Western Australia, Perth, Western Australia 6009, Australia.

Abstract

The method outlined in this article is a customization of the whole exhaust exposure method generated by Mullins et al. (2016) using reprogrammed primary human airway epithelial cells as described by Martinovich et al. (2017). It has been used successfully to generate recently published data (Landwehr et al. 2021). The goal was to generate an exhaust exposure model where exhaust is collected from a modern engine, real-world exhaust concentrations are used and relevant tissues exposed to assess the effects of multiple biodiesel exposures. Exhaust was generated, gently vacuumed into a dilution chamber where it was diluted 1/15 with air and then vacuumed into an incubator containing the primary cell cultures for exposure. Exhaust physico-chemical properties including combustion gas concentrations and particle spectra were then analyzed using a combustion gas analyzer and a Universal Scanning Mobility Particle Sizer. 24 h after exposure, cellular viability and mediator release were measured using Annexin-V/PI staining and meditator multiplexing kits respectively. This method was generated to test biodiesel exhaust exposures but can be easily adapted for any type of engine exhaust exposure or even potentially other respirable environmental exposures such as woodsmoke. The main customization points for this method are:•Exhaust generated by a diesel engine equipped with EURO VI exhaust after treatment devices including diesel particulate filter and diesel oxidation catalyst.•The generated exhaust was diluted 1/15 with air to replicate real world exposure concentrations.•Used primary human airway epithelial cells obtained from bronchoscope brushings from multiple volunteers and reprogrammed to allow multiple, comparative exposures from the same individual.

Keywords: Airway epithelial cells; Biodiesel; Environmental exposure; Exhaust exposure; Exposure protocol.