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ELECTROSTATIC PRECIPITATION AN AN ALTERNATIVE METHOD FOR /IN VITRO/ EXPOSURES TO MIXTURES OF GASES AND PARTICLES
Citation:
DE BRUJINE, K., S. LAKE, K. SEXTON, M. DOYLE, S. EBERVILLER, H. E. JEFFERIES, D. LEITH, AND I. JASPERS. ELECTROSTATIC PRECIPITATION AN AN ALTERNATIVE METHOD FOR /IN VITRO/ EXPOSURES TO MIXTURES OF GASES AND PARTICLES. Presented at Society of Toxicology Annual Meeting, San Deigo, CA, March 05 - 09, 2006.
Description:
There is an increasing interest in examining complex urban air pollution mixtures that include both particulate and gaseous components. Conventional methodologies are unable to expose lung cells in vitro simultaneously to both particulate and gaseous pollutants that are being formed in the ambient air. Therefore, a new method for collection of such mixtures, using electrostatic precipitation, was developed and tested with diesel exhaust (DE). A TSI model 3100 Electrostatic Aerosol Sampler (EAS) was modified to improve the deposition of particles onto respiratory epithelial cells grown on membranes. DE was collected from a 1980 Mercedes-Benz model 300SD, using commercially available diesel fuel, and diluted with room air to a particle volume concentration of approximately 3.0x10^12 nm3/cm3. The particle concentration and size distribution both entering and exiting the EAS were monitored and counted in the range of 13.81 nm to 673 nm. After exposure of respiratory epithelial cells to DE, cells were analyzed for particle deposition and cytotoxicity. The EAS achieved an average collection efficiency of 97% for all particles in the measured size range, which returned to 2% once the EAS was turned off. Cells exposed to the EAS system alone or DE without the EAS showed minimal cytotoxicity and release of IL-8 that was indistinguishable from the incubator controls. However, cells exposed to DE with the EAS turned on produced a threefold increase in LDH and IL-8 release relative to the control. Overall, we were able to directly expose respiratory epithelial cells to DE particles without prior collection in a separate medium, avoiding the possibility of significantly altering the particles¿ characteristics. These results suggest that the EAS system can be used to determine the full toxic potential of both gaseous and particulate components of air pollution mixtures, while also distinguishing the adverse effects of each component separately.
There is an increasing interest in examining complex urban air pollution mixtures that include both particulate and gaseous components. Conventional methodologies are unable to expose lung cells in vitro simultaneously to both particulate and gaseous pollutants that are being formed in the ambient air. Therefore, a new method for collection of such mixtures, using electrostatic precipitation, was developed and tested with diesel exhaust (DE). A TSI model 3100 Electrostatic Aerosol Sampler (EAS) was modified to improve the deposition of particles onto respiratory epithelial cells grown on membranes. DE was collected from a 1980 Mercedes-Benz model 300SD, using commercially available diesel fuel, and diluted with room air to a particle volume concentration of approximately 3.0x10^12 nm3/cm3. The particle concentration and size distribution both entering and exiting the EAS were monitored and counted in the range of 13.81 nm to 673 nm. After exposure of respiratory epithelial cells to DE, cells were analyzed for particle deposition and cytotoxicity. The EAS achieved an average collection efficiency of 97% for all particles in the measured size range, which returned to 2% once the EAS was turned off. Cells exposed to the EAS system alone or DE without the EAS showed minimal cytotoxicity and release of IL-8 that was indistinguishable from the incubator controls. However, cells exposed to DE with the EAS turned on produced a threefold increase in LDH and IL-8 release relative to the control. Overall, we were able to directly expose respiratory epithelial cells to DE particles without prior collection in a separate medium, avoiding the possibility of significantly altering the particles¿ characteristics. These results suggest that the EAS system can be used to determine the full toxic potential of both gaseous and particulate components of air pollution mixtures, while also distinguishing the adverse effects of each component separately.