We developed a cell culture exposure system (CCES) to expose cells at the air-liquid interface (ALI) to volatile chemicals. We characterized the CCES by exposing indigo dye-impregnated filters inside each culture well to 125 ppb ozone (O3) for 1 h at flow rates of 5 and 25 mL/min/well; the reaction of O3 with an indigo dye produces a fluorescence product. We observed a 5-fold increase in fluorescence at 25 mL/min/well, suggesting higher flows were more effective. We then exposed primary human bronchial lung epithelial cells (HBECs) to 0.3 ppm acrolein for 2 h at 3, 5 and 25 mL/min/well and compared our results against well-established Human Studies Facility in vitro exposure chambers (HSF Chambers) at the U.S. EPA. We measured lactate dehydrogenase (LDH) release, and transcript changes of heme oxygenase-1 (HMOX1) and interleukin-8 (IL8) at 0, 1, and 24 h post-exposure. Comparing responses from the HSF Chamber to the CCES, differences were only observed at 1 h post-exposure for HMOX1. Here, the HSF Chamber produced a ~6-fold increase in HMOX1 while the CCES at 3 and 5 mL/min/well produced a ~1.7-fold increase. Operating the CCES at 25 mL/min/well produced a ~4.5-fold increase; slightly lower than the HSF Chamber. Our results suggest that higher flow rates in the CCES were more effective at delivering the gas to the cells, and this was further validated by our comparison against a well-established in vitro exposure system. Further testing is required to explore the sensitivity of the CCES with other chemicals.

This dataset is associated with the following publication: Zavala-Mendez, J., A. Ledbetter, D.S. Morgan, L. Dailey, E. Puckett, S. McCullough, and M. Higuchi. A New Cell Culture Exposure System (CCES) for Studying the Toxicity of Volatile Chemicals at the Air-Liquid Interface. INHALATION TOXICOLOGY. Taylor & Francis, Inc., Philadelphia, PA, USA, 30(4): 169-177, (2018).