Testing of the toxicity of volatile compounds on human lung cells using the Air/Liquid Interface (ALI) culturing and exposure technique: a prevalidation study
2010
Pirow,R. | Smirnova,L. | Liebsch,M. | Tharmann,J. | Luch,A. | Graebsch,C. | Bauer,M. | Linsel,G. | Siemers,R. | Otto,C. | Berger-Preiss,E. | Kock,H. | Oertel,A. | Ritter,D. | Knebel,J.
Alemán. Testing requirements under the new European chemicals policy (REACH) are expected to trigger a large number of toxicologicalstudies. For environmentally and occupationally relevant substances which are taken up by inhalation this will prompt extensive efforts in terms of toxicological assessments unlikelyto be realised through in vivo testing alone. The aim of the present study was to perform a multi-laboratory evaluation of an Air/Liquid Interface (ALI) culturing and exposure technique for testing the cytotoxic and genotoxic effects of inhalable substances(gases) on human lung cells.Human lung cells of the alveolar cell line A549 were grown on microporous membranes at the air/liquid interface and wereexposed to test atmospheres consisting of different mixtures of synthetic air supplemented with the test gas (i.e. NO2, SO2, formaldehydeor ozone). Gas-mediated cytotoxicity and genotoxicity were assessed via electronic cell counting (CASY technology) and the COMET assay, respectively. Logistic regressionand hierarchical modelling were employed to determine the intra- and interlaboratory variabilities in dose-response relationships.A linear-regression prediction model was developed for the cytotoxic endpoint.Analyses of dose-response relationships for the endpoint cell number showed a good repeatability within and reproducibility between the laboratories for all four gases. Comparison of thederived EC50 values with published LC50 values for mice and rats revealed a tight quantitative relationship between in vitro cytotoxicity and in vivo lethality. Further, analyses of the tail-momentsobtained from the COMET assay demonstrated clear and reproducible dose-response relationships for SO2 and formaldehyde, being indicative for dose-dependent DNA strand breaks (SO2)and DNA-protein crosslinks (formaldehyde). No such dose-dependent effects could be observed for NO2 and ozone.The results of the present prevalidation study are promising with respect to the reliability and relevance of the proposed in vitro method for inhalation toxicity testing. Before entering aformal validation stage, extended prevalidation will be necessary to establish a tested training set of compounds sufficiently large to allow for optimisation of the prediction model
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Este registro bibliográfico ha sido proporcionado por German Federal Institute for Risk Assessment