Cytotoxicity and reactive oxygen species generation from aggregated carbon and carbonaceous nanoparticulate materials

Kristine M Garza¹, Karla F Soto², Lawrence E Murr³

¹Department of Biological Sciences, The University of Texas at El Paso, El Paso, TX, USA; ²Lockheed Martin Aeronautics Company, Forth Worth, TX, USA; ³Department of Metallurgical and Materials Engineering, The University of Texas at El Paso, El Paso, TX, USA

Abstract: We have investigated the cytotoxicity and reactive oxygen species (ROS) generation for indoor and outdoor soots: candle, wood, diesel, tire, and natural gas burner soots – along with surrogate black carbon, various multiwall carbon nanotube aggregate materials, TiO2 (anatase) and chrysotile asbestos as reference materials. All soots were observed utilizing TEM and FESEM to be composed of aggregated, primary spherules (20–80 nm diameter) forming complex, branched fractal structures. These spherules were composed of intercalated, turbostratic arrangements of curved graphene fragments with varying concentrations of polycyclic aromatic hydrocarbon (PAH) isomers. In vitro cultures with an immortalized human lung epithelial carcinoma cell line (A549) treated with these materials showed decreased cell viability and variations in ROS production, with no correlations to PAH content. The data demonstrate that soots are cytotoxic and that cytotoxicity is not related to PAH content but is related to ROS generation, suggesting that soot induces cellular oxidative stress and that cell viability assays can be indicators of ROS production.

Keywords: cytotoxicity assessment, ROS assays, FESEM and TEM analysis, nanoparticulate aggregates