| Title: | A new cell culture exposure system for studying the toxicity of volatile chemicals at the air-liquid interface |
| Author(s): | Zavala J; Ledbetter AD; Morgan DS; Dailey LA; Puckett E; McCullough SD; Higuchi M; |
| Address: | "a NHEERL , U.S. Environmental Protection Agency, Research Triangle Park , NC , USA. b NHEERL, U.S. Environmental Protection Agency , Chapel Hill , NC , USA" |
| DOI: | 10.1080/08958378.2018.1483983 |
| ISSN/ISBN: | 1091-7691 (Electronic) 0895-8378 (Print) 0895-8378 (Linking) |
| Abstract: | "A cell culture exposure system (CCES) was developed to expose cells established at an air-liquid interface (ALI) to volatile chemicals. We characterized the CCES by exposing indigo dye-impregnated filter inserts inside culture wells to 125 ppb ozone (O(3)) for 1 h at flow rates of 5 and 25 mL/min/well; the reaction of O(3) with an indigo dye produces a fluorescent product. A 5-fold increase in fluorescence at 25 mL/min/well versus 5 mL/min/well was observed, suggesting higher flows were more effective. We then exposed primary human bronchial 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 in vitro exposure chambers at the U.S. EPA's Human Studies Facility (HSF Chambers). We measured transcript changes of heme oxygenase-1 (HMOX1) and interleukin-8 (IL-8), as well as lactate dehydrogenase (LDH) release, at 0, 1, and 24 h post-exposure. Comparing responses from HSF Chambers to the CCES, differences were only observed at 1 h post-exposure for HMOX1. Here, the HSF Chamber produced a approximately 6-fold increase while the CCES at 3 and 5 mL/min/well produced a approximately 1.7-fold increase. Operating the CCES at 25 mL/min/well produced a approximately 4.5-fold increase; slightly lower than the HSF Chamber. Our biological results, supported by our comparison against the HSF Chambers, agree with our fluorescence results, suggesting that higher flows through the CCES are more effective at delivering volatile chemicals to cells. This new CCES will be deployed to screen the toxicity of volatile chemicals in EPA's chemical inventories" |
| Keywords: | "Acrolein/*toxicity Biomarkers/metabolism Bronchi/*drug effects/metabolism/pathology Cells, Cultured Epithelial Cells/*drug effects/metabolism/pathology Heme Oxygenase-1/genetics/metabolism Humans Inhalation Exposure Interleukin-8/genetics/metabolism L-Lac;" |
| Notes: | "MedlineZavala, Jose Ledbetter, Allen D Morgan, David S Dailey, Lisa A Puckett, Earl McCullough, Shaun D Higuchi, Mark eng EPA999999/Intramural EPA/ Research Support, U.S. Gov't, Non-P.H.S. England 2018/08/09 Inhal Toxicol. 2018 Mar-Apr; 30(4-5):169-177. doi: 10.1080/08958378.2018.1483983. Epub 2018 Aug 8" |
