Design and Testing of Electrostatic Aerosol<i>In Vitro</i>Exposure System (EAVES): An Alternative Exposure System for Particles

Conventional in vitro exposure methods for cultured human lung cells rely on prior suspension of particles in a liquid medium; these have limitations for exposure intensity and may modify the particle composition. Here electrostatic precipitation was used as an effective method for such in vitro exposures. An obsolete electrostatic aerosol sampler was modified to provide a viable environment within the deposition field for human lung cells grown on membranous support. Particle deposition and particle-induced toxicological effects for a variety of particles including standardized polystyrene latex spheres (PSL) and diesel exhaust emission particle mixtures are reported. The Electrostatic Aerosol in Vitro Exposure System (EAVES) efficiently deposited particles from an air stream directly onto cells. Cells exposed to the electric field of the EAVES in clean air or in the presence of charged PSL spheres exhibited minimal cytotoxicity, and their release of inflammatory cytokines was indistinguishable from that of the controls. For the responses tested here, there are no significant adverse effects caused neither by the electric field alone nor by the mildly charged particles. Exposure to diesel exhaust emissions using the EAVES system induced a threefold increase in cytokines and cytotoxicity as compared to the control. Taken together, these data show that the EAVES can be used to expose human lung cells directly to particles without prior collection in media, thereby providing an efficient and effective alternative to the more conventional particle in vitro exposure methods.