Prevalidation of an Acute Inhalation Toxicity Test Using the EpiAirway In Vitro Human Airway Model

George R. Jackson, Jr., Anna G. Maione, Mitchell Klausner, and Patrick J. Hayden

Introduction: Knowledge of acute inhalation toxicity potential is important for establishing safe use of chemicals and consumer products. Inhalation toxicity testing and classification procedures currently accepted within worldwide government regulatory systems rely primarily on tests conducted in animals. The goal of the current work was to develop and prevalidate a nonanimal (in vitro) test for determining acute inhalation toxicity using the EpiAirway™ in vitro human airway model as a potential alternative for currently accepted animal tests. Materials and Methods: The in vitro test method exposes EpiAirway tissues to test chemicals for 3 hours, followed
by measurement of tissue viability as the test endpoint. Fifty-nine chemicals covering a broad range of toxicity classes, chemical structures, and physical properties were evaluated. The in vitro toxicity data were utilized
to establish a prediction model to classify the chemicals into categories corresponding to the currently accepted Globally Harmonized System (GHS) and the Environmental Protection Agency (EPA) system. Results: The EpiAirway prediction model identified in vivo rat-based GHS Acute Inhalation Toxicity Category 1–2 and EPA Acute Inhalation Toxicity Category I–II chemicals with 100% sensitivity and specificity of 43.1% and 50.0%, for GHS and EPA acute inhalation toxicity systems, respectively. The sensitivity and specificity of the EpiAirway prediction model for identifying GHS specific target organ toxicity-single exposure (STOT-SE) Category 1 human toxicants were 75.0% and 56.5%, respectively. Corrosivity and electrophilic and oxidative reactivity appear to be the predominant mechanisms of toxicity for the most highly toxic chemicals. Conclusions: These results indicate that the EpiAirway test is a promising alternative to the currently accepted animal tests for acute inhalation toxicity.


acute inhalation toxicity, EpiAirway (AIR-100), Globally Harmonized System (GHS) classification, Environmental Protection Agency (EPA) classification, Millicell-CM-cap, IC75

Materials Tested

dimethylsulfoxide (DMSO), ethanol, acetone, corn oil, olive oil, 1-Butanol, 2-Butoxyethanol, 2-Ethoxyethanol, 2-Ethoxyethyl acetate, 4-Ethylmorpholine, Acetic acid, Acetone, Acetonitrile, Acrolein, Allyl alcohol, Allyl chloride, Allyl glycidyl ether, Aniline, Benzene, Benzoyl chloride, Benzyl chloride, Butylamine, Carbon Tetrachloride, Chloroacetaldehyde, Chloroform, Crotonaldehyde, Cyclohexane, Cyclohexanol, Cyclohexanone, Dichloroethyl ether, Diethyl(amino)ethanol, Diethylamine, Diisopropylamine, Dimethylacetamide, Dimethylamine, Dimethylphthalate, Ethanolamine, Ethyl acrylate, Ethyl alcohol, Ethyl formate, Ethylene chlorohydrins, Formaldehyde, Formic acid, Glycidol, Hydrogen peroxide, Hexane, Hexone, Isophorone, Methyl acrylate, Methyl Isobutyl Ketone, Methyl methacrylate, Morpholine, N,N-Dimethylformamide, n-Heptane, p-Anisidine, Paraquat, p-Dichlorobenzene, Phenol, Propylene Glycol, p-tert-Butyltoluene, p-Xylene, Tetrachloroethylene, Trichloroethylene, Triethylene glycol, Vinyl Acetate

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