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Hanging drop: An in vitro air toxic exposure model using human lung cells in 2D and 3D structures

Liu, F. F. and Peng, C. and Escher, B. I. and Fantino, E. and Giles, C. and Were, S. and Duffy, L. and Ng, J. C. (2013) Hanging drop: An in vitro air toxic exposure model using human lung cells in 2D and 3D structures. Journal of Hazardous Materials, 261 . pp. 701-710. ISSN 0304-3894

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Article Link(s): http://dx.doi.org/10.1016/j.jhazmat.2013.01.027

Publisher URL: http://www.sciencedirect.com/science/article/pii/S0304389413000459

Abstract

Using benzene as a candidate air toxicant and A549 cells as an in vitro cell model, we have developed and validated a hanging drop (HD) air exposure system that mimics an air liquid interface exposure to the lung for periods of 1 h to over 20 days. Dose response curves were highly reproducible for 2D cultures but more variable for 3D cultures. By comparing the HD exposure method with other classically used air exposure systems, we found that the HD exposure method is more sensitive, more reliable and cheaper to run than medium diffusion methods and the CULTEX (R) system. The concentration causing 50% of reduction of cell viability (EC50) for benzene, toluene, p-xylene, m-xylene and o-xylene to A549 cells for 1 h exposure in the HD system were similar to previous in vitro static air exposure. Not only cell viability could be assessed but also sub lethal biological endpoints such as DNA damage and interleukin expressions. An advantage of the HD exposure system is that bioavailability and cell concentrations can be derived from published physicochemical properties using a four compartment mass balance model. The modelled cellular effect concentrations EC50(cell) for 1 h exposure were very similar for benzene, toluene and three xylenes and ranged from 5 to 15 mmol/kg(dry weight) which corresponds to the intracellular concentration of narcotic chemicals in many aquatic species, confirming the high sensitivity of this exposure method. (C) 2013 Elsevier B.V. All rights reserved.

Item Type:Article
Business groups:Biosecurity Queensland
Additional Information:Liu, Faye F. Peng, Cheng Escher, Beate I. Fantino, Emmanuelle Giles, Cindy Were, Stephen Duffy, Lesley Ng, Jack C. Crccare [1-3-03-07/08]; crccare We wish to acknowledge the guidance of Dr. Nicolas Timmons in 3D HD cell culturing techniques; QHFSS Histology Laboratory and the Cytology Laboratory in Queen Elizabeth It Hospital for spheroid cell block making and sample embedding (these results are supplied in online supplementary document) and Nat Ling Jin for the help of graph productions. One plate of 12 free samples of Alvetex (R) (Reinnervate # AVP002) inserts and 3 samples of 6 welled 3D inserts (AVP004-3) were kindly provided by Bio Scientific. This project was funded by CRCCARE (project No.: 1-3-03-07/08). A PhD scholarship to F.F. Liu was also provided by CRCCARE. Entox is a partnership between Queensland Health and the University of Queensland. Elsevier science bv Amsterdam
Keywords:Hanging drop In vitro air exposure Lung cells Benzene Bioavailability multicellular tumor spheroids organic-compounds epithelial-cells gaseous compounds cultured-cells ozone exposure system chemicals a549 metabolism
Subjects:Science > Biology > Cytology
Deposited On:16 Jul 2014 03:01
Last Modified:16 Jul 2014 03:01

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