Exposure of 19 substances to lung A549 cells at the air liquid interface or under submerged conditions reveals high correlation between cytotoxicity in vitro and CLP classifications for acute lung toxicity

Abstract

In vivo experiments are still widely used for the testing of lung toxicity but there is an ethical and legal obligation to replace, reduce and refine animal testing. Lung A549 cells could serve as an in vitro indicator for acute lung toxicity but little data about the correlation of the cytotoxicity in A549 cells and data leading to CLP classifications are available. We exposed A549 cells to 19 CLP-classified substances with doses of 25, 50, and 100 μg/cm² either under submerged (SME) condition or with aerosols at the air-liquid interface (ALIF) and determined accuracy, precision, sensitivity and the F1 score with the CLP classifications H330, H332, or H335. When data from both exposure methods were combined, we found accuracies of 0.84 ± 0.05, precisions of 0.74 ± 0.1, sensitivities of 0.93 ± 0.08 and F1 scores of 0.82 ± 0.04. Separated from each other, ALIF exposure was more sensitive at any dose but, at higher doses, also less accurate and precise compared to SME. Considering the 19 substances tested, our data suggest that cytotoxicity in A549 cells could be a reliable in vitro indicator for in vivo toxicity. Thus, we discuss how A549 could be integrated into validation test guidelines.

Keywords: A549; Acute lung toxicity; Air-liquid; Laboratory animal; Regulation; Submerged.