Development of an in vitro Batch-type Closed Gas Exposure Device with an Alveolar Epithelial Cell Line, A549, for Toxicity Evaluations of Gaseous Compounds

Abstract

To simply evaluate toxicity for various types of exhaust-gas samples collected in various locations, we developed a smallscale (150 mL) batch-type completely closed gas exposure device incorporated with an air-liquid interface culture of a human alveolar epithelial cell line, A549. On the basis of cell viability tests using an acid phosphatase assay after 48 h of gas exposure, the developed device was able to measure clear dose-response relationships for volatile organic and inorganic compounds, such as benzene, trichloroethylene (TCE), acetone, SO2 and NO2 gases, but not CO gas. Although the 50% effective concentration values in the device were much higher than 50% lethal concentration values reported in animal experiments, the tendency of the toxic intensity observed in the former was roughly consistent with that of the acute toxicity in the latter. We further applied the device to evaluate the toxicity of cigarette smoke as an example of actual environmental gases, and successfully measured acute cell death from the gas after 48 h of exposure. The present small device is expected to be one of good tools not only in simultaneously assessing various gaseous chemicals or samples, but also in studying acute toxicity expression mechanisms in human lung epithelia.

Realistic Exposure Methods for Investigating the Interaction of Nanoparticles with the Lung at the Air-Liquid Interface In Vitro

“…The contact of the exhaust emissions occurs over diffusion, as it is also the case in the gas exchange parts of the human airways (79). In comparison to other such exposure systems (Table 1) (97)(98)(99)(100)(101)(102)(103)(104)(105)(106), as also highlighted in (38), the control of the conditions, the dilution rate and the exchange of diffusion processes are more realistic in the exposure system of Muller et al (38).…”

An Alternative Gas-phase In Vitro Exposure System for Toxicity Testing: The Interaction Between Nitrous Oxide and A549 Cells

“…N 2 O was selected as the volatile compound, since it is commonly used as an adjunct to general anaesthesia. The A459 adenocarcinoma cell line (type II pneumocytes) has been widely used for investigations on in vitro exposure to volatile compounds (5,6,9,13,14,25,26). Besides this application, these cells have been used to explore the inflammatory effects of volatile compounds by using an ALIC system, in vitro.…”

“…Their main limitation is that there is no assurance that all the cell culture plates are exposed to the same concentration of the test compound. Recently, two similar air-liquid interface cultures (ALICs) have been developed, which enable cells to make contact with gases directly at the apical side, while the cells are fed with medium from the basal side through a semi-permeable membrane, all within a closed gas chamber (6,13,14).…”

Safety Evaluation of Dry Powder Formulations by Direct Dispersion onto Air–Liquid Interface Cultured Cell Layer

“…[19][20][21][22][23][24] In this study, thus, to apply ALI to evaluation of the safety of dry powders, we compared the safety of several candidate excipients with two cell culture systems (ALI with Transwell ® insert and SUB with a 96 well plate and Transwell ® insert)…”