Alternative air–liquid interface method for inhalation toxicity testing of a petroleum-derived substance

October 8, 2020

Verstraelen Sandra a , Jacobs An a , Van Laer Jo a , Van Deun Masha b , Bertels Diane b , Hilda Witters a , Remy Sylvie a , c , Geerts Lieve a , Deferme Lize d , Frijns Evelien a
a VITO NV (Flemish Institute for Technological Research), Unit HEALTH, Mol, Belgium 
b VITO NV, Unit SCT (Separation and Conversion Technology), Mol, Belgium 
c Department of Biomedical Sciences, University of Antwerp, Antwerp, Belgium 
d ExxonMobil Petroleum and Chemical B.V., Machelen, Belgium


A549 cells were exposed to gasoline at the ALI by passive dosing, an approach that is already used in aquatic toxicity testing. This showes a clear dose-dependent biological response. This in vitro -based new approach methodology might be promising for inhalation toxicity testing of (semi-)volatile complex substances.


In vitro -based new approach methodologies (NAMs) provide a pragmatic solution to animal testing of petroleum substances and their constituents. A previous study exposed an in vitro model (A549 cells) at the air–liquid interface (ALI) to assess inhalation toxicity of a single compound, ethylbenzene. Experimental conditions using VITROCELL® 24/48 exposure system were optimized to achieve a deposition efficiency that resulted in dose- dependent biological changes. The feasibility of this set-up was evaluated for testing the complex substance gasoline, which, at only high concentrations, can induce mild respiratory irritation in animals and cough in humans.

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