Assessment of Biological Responses of EpiAirway™ 3-D Cell Constructs vs. A549 Cells for Determining Toxicity of Ambient Air Pollution

Jose Zavala, Bridget O’Brien, Kim Lichtveld, Kenneth G. Sexton, Ivan Rusyn, Ilona Jaspers and William Vizuete

Context — EpiAirway™ 3-D constructs are human-derived cell cultures of differentiated airway epithelial cells that may represent a more biologically relevant model of the human lung. However, limited information is available of its utility for exposures to air pollutants at the air-liquid interface (ALI). Objective — To assess the biological responses of EpiAirway™ cells in comparison to the responses of A549 human alveolar epithelial cells after exposure to air pollutants at ALI. Methods — Cells were exposed to filtered air, 400ppb of ozone (O3) or a photochemically-aged Synthetic Urban Mixture (SynUrb54) consisting of hydrocarbons, nitrogen oxides, O3, and other secondary oxidation products for 4 h. Basolateral supernatants and apical washes were collected at 9 and 24 h post-exposure. We assessed cytotoxicity by measuring lactate dehydrogenase (LDH) release into the culture medium and apical surface. Interleukin 6 (IL-6) and interleukin 8 (IL-8) proteins were measured in the culture medium and in the apical washes to determine the inflammatory response after exposure. Results — Both O3 and SynUrb54 significantly increased basolateral levels of LDH and IL-8 in A549 cells. No significant changes in LDH and IL-8 levels were observed in the EpiAirway™ cells, however, IL-6 in the apical surface was significantly elevated at 24 h after O3 exposure. Conclusion — LDH and IL-8 are robust endpoints for assessing toxicity in A549 cells. The EpiAirway™ cells show minimal adverse effects after exposure suggesting that they are more toxicologically resistant compared to A549 cells. Higher concentrations or longer exposure times are needed to induce effects on EpiAirway™ cells


EpiAirway (AIR-100-SNP), A549 human alveolar epithelial cells, LDH, IL-6, IL-8, Inhalation toxicology, environmental irradiation chamber, air pollution

Materials Tested

ozone (O3), Synthetic Urban Mixture (SynUrb54) - photochemically-aged, nitric oxide (NO), nitrogen dioxide (NO2)

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