Comparing the Biocompatibility of Diffusion Flame-Generated Soot to Human Lung Cells by Two Commonly Used In-Vitro Cultures: Monolayer and Air-Liquid Interface
Durgesh N. Das, Paul Hamilton, Shubham Sunil Sharma, DHRUV MITROO, Benjamin Kumfer, Veterans Research and Education Foundation
Abstract Number: 556
Working Group: Health-Related Aerosols
Abstract
In-vitro models are the avenues by which researchers evaluate initial biological responses and discover cellular pathways of mammalian tissues upon exposure to environmental pollutants. Perhaps the most well-recognized airborne environmental pollutant, soot, accounts for many pathologies associated with aerosol exposure. At present, several in-vitro models are employed to address similar questions, ranging from cell types to culturing conditions, because not everyone has access to latest technology and methodology.
To this end, we compare initial biological responses and some hallmark features of human pulmonary cells to soot, grown using two in-vitro methods: traditional monolayer coverage and the organ tissue equivalent (OTE) cultured the air-liquid interface (ALI). These are both not novel techniques, however, to our knowledge, the two most employed by researchers with roots in aerosol science. We treat human small airway epithelial cells (HSAEC; a commercially available primary cell line), cultured to provide both models, with laboratory-generated diffusion flame soot. Of the biological responses, we compare LD50, oxidative stress, and mitochondrial damage and dysfunction. To obtain insight into the cellular pathways occurring in both models, we monitor for makers of necrosis vs apoptosis. This is complemented with immunohistochemistry staining and immunofluorescent staining (e.g., for cleaved Caspase 3 during apoptosis). We hope this can allow a more direct comparison across studies that employ both models, current and prospective, and/or reveal any obvious discrepancies.