Genotoxicity induced by fine urban air particulate matter in the macrophages cell line RAW 264.7

Recent studies support a participation of fine airborne particulate matter (PM) with an aerodynamic diameter less than 2.5 microm in the effects of air pollutants on health. Particulate matter was collected in an urban area of L'Aquila during the winter 2004. Fine particulate samples were analyzed by X-ray photoelectron spectroscopy (XPS) to determine the chemical inventory of the aerosol particle surfaces and to evaluate the weight of characteristic functional groups of the most frequent carbon-containing organic pollutant compounds (C-C/C-H, C-O/C-N, C=O, COOH). The most important contributor to the mass of fine particulate matter was carbon. The overall purpose of this work was to determine the in vitro toxicity and genotoxicity of fine PM in cultured macrophages (RAW 264.7 cells) since the biological target of inhaled PM are the pulmonary epithelium and resident macrophages. In parallel in vitro toxicity assays were used including cell viability and apoptosis. Genotoxicity was evaluated by the micronucleus (MN) assay. The viability of macrophages was assessed by the MTT method; apoptosis by an ELISA test for programmed cell death (PCD) was determined after RAW 264.7 cells treatment. Concentration of 1, 3 and 10 microg/cm2 of fine particles induced micronuclei in a dose-dependent manner. We also compared the effects of fine PM with those of fine carbon black particles (CB) in similar doses. Fine carbon black particles were consistently less genotoxic than the fine atmospheric particles, suggesting that the contaminants adsorbed on them (i.e. carbon-containing organic compounds in addition to metal oxides and metal salts) are involved in genotoxicity. Fine PM reduced cellular proliferation. Overall, the results presented here demonstrate the utility of in vitro tests in mouse cells for testing genotoxicity of urban air particulate matter.