In Vitro primary human airway epithelial whole exhaust exposure

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.

Respiratory Health Effects of In Vivo Sub-Chronic Diesel and Biodiesel Exhaust Exposure

“…Both Canola and Tallow biodiesel were created following an established sodium methoxide transesterification process [ 102 ] using high-quality oils obtained from local suppliers (Campbells Wholesale Reseller, WA, Aus and Range Products, WA, Aus). Detailed FAME profiles have been previously published [ 54 , 103 ]. The diesel exhaust exposure consumed less fuel due than was required for both biodiesel exposures, likely due to the differences in fuel efficiency [ 18 ].…”

“…The exposure methodology is based on a combination of previously published protocols [ 41 , 103 ]. To make exposures more realistic to occupational settings, mice were exposed for short time periods to exhaust diluted approximately 1/10 with air with cold-start emissions included as part of the exposure.…”

Biodiesel Exhaust Toxicity with and without Diethylene Glycol Dimethyl Ether Fuel Additive in Primary Airway Epithelial Cells Grown at the Air–Liquid Interface

“…The exposure method followed a previously published protocol . Briefly, each exposure was done on a different day to control for cold start effects.…”

“…The exposure method followed a previously published protocol. 32 Briefly, each exposure was done on a different day to control for cold start effects. The exhaust was generated using a single-cylinder, 435cc design Yanmar L100V engine (Yanmar, Italy), fitted with EURO V/VI diesel exhaust after-treatment equipment including a diesel particulate filter (DPF) and diesel oxidation catalyst (DOC, both Daimler, Germany), coupled with a dynamometer.…”