Toward Standardization of a Lung New Approach Model for Toxicity Testing of Nanomaterials.

Abstract

This study represents an attempt toward the standardization of pulmonary NAMs and the development of a novel approach for toxicity testing of nanomaterials. Laboratory comparisons are challenging yet essential for identifying existing limitations and proposing potential solutions. Lung cells cultivated and exposed at the air-liquid interface (ALI) more accurately represent the physiology of human lungs and pulmonary exposure scenarios than submerged cell and exposure models. A triculture cell model system was used, consisting of human A549 lung epithelial cells and differentiated THP-1 macrophages on the apical side, with EA.hy926 endothelial cells on the basolateral side. The cells were exposed to silver nanoparticles NM-300K for 24 h. The model used here showed to be applicable for assessing the hazards of nanomaterials and chemicals, albeit with some limitations. Cellular viability was measured using the alamarBlue assay, DNA damage was assessed with the enzyme-modified comet assay, and the expression of 40 genes related to cell viability, inflammation, and DNA damage response was evaluated through RT² gene expression profiling. Despite harmonized protocols used in the two independent laboratories, however, some methodological challenges could affect the results, including sensitivity and reproducibility of the model.