AAAR 34th Annual Conference
October 12 - October 16, 2015
Hyatt Regency
Minneapolis, Minnesota, USA
Abstract View
Cellular Assays for Measuring Reactive Oxygen Species (ROS) Production of Particulate Matter Mixtures
WING-YIN TUET, Vishal Verma, Meghan Knight, Julie Champion, Anna Grosberg, Nga Lee Ng, Georgia Institute of Technology
Abstract Number: 445 Working Group: Health Related Aerosols
Abstract The health effects of particulate matter (PM) have received considerable attention with an increasing number of epidemiological studies associating PM exposure with increases in cardiovascular and respiratory diseases. Furthermore, the World Health Organization (WHO) has classified PM as a Group I carcinogen, meaning there is sufficient evidence that it causes cancer in humans. Despite these findings, the mechanism by which PM exposure results in adverse health effects is unclear. Toxicology studies have suggested that PM-induced reactive oxygen species (ROS) production can initiate inflammatory cascades and that prolonged stimulation of these cascades can lead to oxidative stress and cellular damage. This provides a possible link between PM exposure and adverse health effects as PM can directly generate ROS and induce cellular pathways that produce ROS.
Here we present ROS measurements of ambient PM samples collected in the greater Atlanta area as part of the Southeastern Center for Air Pollution and Epidemiology (SCAPE) study. Murine alveolar macrophages and neonatal rat ventricular myocytes were used to measure cellular oxidative stress via particle-induced ROS production using a fluorescent probe. These cell types were chosen to represent the respiratory and cardiovascular system respectively. Briefly, the probe is non-fluorescent until it reacts with ROS and the measured fluorescence is proportional to ROS produced. For each cell type, various assay parameters (cell number, sample incubation time, etc.) were optimized to give the maximum signal separation between positive and negative controls. Finally, after optimization, filter dose responses were obtained by measuring ROS production in a series of dilutions for filters collected at rural (Yorkville) and urban (Jefferson Street) sites to compare ROS activity and composition between the two.