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Cellular response to the deposition of diesel exhaust particle aerosols onto human lung cells grown at the air–liquid interface by inertial impaction
Authors:Daniel J. Cooney  Anthony J. Hickey
Affiliation:aDepartment of Biomedical Engineering, University of North Carolina, Chapel Hill, NC, USA;bEshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC, USA
Abstract:The pathogenesis of disease resulting from exposure to diesel exhaust particles (DEP) is often studied using cultured lung cells. Frequently, researchers expose cells to DEP by spiking a suspension of particles in liquid onto the apical surface. This is not representative of in vivo exposure, where aerosols are deposited onto cell surfaces at the air–liquid interface (ALI). Inertial impaction provides an opportunity to deliver high doses of particles with aerodynamic diameters >∼1 μm to the surface of cells in seconds in a reproducible and predictable manner.A custom device was constructed to deposit DEP aerosols onto the surface of Calu-3 and A549 cells grown at the ALI. The pro-inflammatory and toxic cellular response to exposure to the deposited DEP aerosols was measured and compared to the response of cells exposed to DEP as suspensions. Calu-3 cells showed evidence of an oxidative stress response for both exposure types, while there was strong evidence to suggest that the method of aerosol delivery was harmful to the A549 cells.
Keywords:Abbreviations: ALI, air&ndash  liquid interface   ANOVA, analysis of variance   CDC, custom deposition chamber   d50, 50% aerodynamic cutoff diameter   da, aerodynamic diameter   DEP, diesel exhaust particles   DF, disodium fluorescein   ELISA, enzyme-linked immunosorbent assay   GM-CSF, granulocyte macrophage-colony stimulating factor   HFA134a, hydrofluoroalkane 134a   IL-8, interleukin-8   LDH, lactate dehydrogenase   Papp, apparent permeability   PBS, phosphate buffered saline   ROS, reactive oxygen species   SEM, scanning electron microscopy   TSLI, twin-stage liquid impinger
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