Samuel Chung 1 2, Nathalie Baumlin 1 2, John S Dennis 1 2, Robert Moore 2, Sebastian F Salathe 2, Phillip L Whitney 2, Juan Sabater 3, William M Abraham 3, Michael D Kim 1 2, Matthias Salathe 1 2
1Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, University of Kansas Medical Center, Kansas City, Kansas.
2Division of Pulmonary, Critical Care and Sleep Medicine, University of Miami School of Medicine, Miami, Florida; and.
3Department of Research, Mount Sinai Medical Center, Miami Beach, Florida.
The transient receptor potential ankyrin 1 (TRPA1) is a molecular target for vape effects due to its expression in airway epithelia and its reported gating by nicotine, reactive oxidants, and flavors, especially cinnamaldehyde. To test whether nicotine had effects independent of other e-cig vapor constituents, the Vitrocell® CLOUD exposure system was utilized to nebulize fixed nicotine doses onto the apical surface of ALI cultures. A549 cell cultures were exposed to nicotine containing e-cig vapor, produced by the VC-1 smoke exposure robot, in the air-liquid interface.
Rationale: Electronic cigarette (e-cig) use has been widely adopted under the perception of safety. However, possibly adverse effects of e-cig vapor in never-smokers are not well understood.Objectives: To test the effects of nicotine-containing e-cig vapors on airway mucociliary function in differentiated human bronchial epithelial cells isolated from never-smokers and in the airways of a novel, ovine large animal model.Methods: Mucociliary parameters were measured in human bronchial epithelial cells and in sheep. Systemic nicotine delivery to sheep was quantified using plasma cotinine levels, measured by ELISA.Measurements and Main Results: In vitro, exposure to e-cig vapor reduced airway surface liquid hydration and increased mucus viscosity of human bronchial epithelial cells in a nicotine-dependent manner. Acute nicotine exposure increased intracellular calcium levels, an effect primarily dependent on TRPA1 (transient receptor potential ankyrin 1). TRPA1 inhibition with A967079 restored nicotine-mediated impairment of mucociliary parameters including mucus transport in vitro. Sheep tracheal mucus velocity, an in vivo measure of mucociliary clearance, was also reduced by e-cig vapor. Nebulized e-cig liquid containing nicotine also reduced tracheal mucus velocity in a dose-dependent manner and elevated plasma cotinine levels. Importantly, nebulized A967079 reversed the effects of e-cig liquid on sheep tracheal mucus velocity.Conclusions: Our findings show that inhalation of e-cig vapor causes airway mucociliary dysfunction in vitro and in vivo. Furthermore, they suggest that the main nicotine effect on mucociliary function is mediated by TRPA1 and not nicotinic acetylcholine receptors.