The biological effects induced by complete engine emissions in a 3D model of the human airway (MucilAirTM) and in human bronchial epithelial cells (BEAS-2B) grown at the air-liquid interface were compared. The cells were exposed for one or five days to emissions generated by a Euro 5 direct injection spark ignition engine. The general condition of the cells was assessed by the measurement of transepithelial electrical resistance and mucin production. The cytotoxic effects were evaluated by adenylate kinase (AK) and lactate dehydrogenase (LDH) activity. Phosphorylation of histone H2AX was used to detect double-stranded DNA breaks. The expression of the selected 370 relevant genes was analyzed using next-generation sequencing. The exposure had minimal effects on integrity and AK leakage in both cell models. LDH activity and mucin production in BEAS-2B cells significantly increased after longer exposures; DNA breaks were also detected. The exposure affected CYP1A1 and HSPA5 expression in MucilAirTM. There were no effects of this kind observed in BEAS-2B cells; in this system gene expression was rather affected by the time of treatment. The type of cell model was the most important factor modulating gene expression. In summary, the biological effects of complete emissions exposure were weak. In the specific conditions used in this study, the effects observed in BEAS-2B cells were induced by the exposure protocol rather than by emissions and thus this cell line seems to be less suitable for analyses of longer treatment than the 3D model.

Center of Vehicles for Sustainable Mobility Faculty of Mechanical Engineering Czech Technical University Prague Technicka 4 160 00 Prague Czech Republic

Department of Chemistry and Toxicology Veterinary Research Institute 621 00 Brno Czech Republic

Department of Computer Science Czech Technical University Prague 12135 Prague Czech Republic

Department of Genetic Toxicology and Nanotoxicology Institute of Experimental Medicine of the CAS Videnska 1083 142 20 Prague Czech Republic

Department of Physiology Faculty of Science Charles University Vinicna 7 128 44 Prague Czech Republic

Department of Vehicles and Ground Transport Czech University of Life Sciences Prague Kamycka 129 165 21 Prague Czech Republic

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