AAAR 35th Annual Conference October 17 - October 21, 2016 Oregon Convention Center Portland, Oregon, USA
Abstract View
Improved in vitro Inhalation Toxicology Method for Nanoparticles and Combustion Emissions
PASI, I JALAVA, Kari Kuuspalo, Mika Ihalainen, Oskari Uski, Tuukka Ihantola, Olli Sipppula, Jarkko Tissari, Jorma Jokiniemi, Maija-Riitta Hirvonen, University of Eastern Finland
Abstract Number: 294 Working Group: Health Related Aerosols
Abstract Air liquid interface (ALI) methods have been introduced to improve the accuracy of aerosol toxicology studies. However, particle deposition in the ALI methods have been least effective with the particles <100nm in aerodynamic diameter. In our new system, thermophoresis has been applied to increase deposition of the particles in combustion emission and, thus, reliably study the toxicological effects of emissions and nanoparticles (Ihalainen et al. AAAR abstract).
We used the new exposure device in exposing human alveolar A549 cells to wood combustion aerosol. The aerosol was measured for the gaseous and particulate compounds. The cells were cultured at the membrane inserts for four to six days to form an epithelial barrier at air liquid interface. The cells were exposed at air liquid interface for one hour to diluted wood combustion aerosol from a stove operated with the batches of beech wood logs. We also studied the effects of aged wood smoke. After the one hour exposures, cell culture inserts were moved to the support plates and allowed to recover in an incubator for 24 hours. Thereafter, toxicological endpoints were measured including fluorescence imaging of the cells in the inserts. The other toxicity endpoints using detached cells, included analyses of thiol reduction, mitochondrial potential, cellular metabolic activity and oxidative stress. In addition, the cell culture mediums were analyzed for cytokine concentration.
The new ALI exposure device showed to be well suited for combustion emission toxicology studies, in addition to nanoparticles described in the other abstract. The constantly good viability of the clean air exposed control cells provides good repeatability of the studies and reduces artifacts in the comparability between the clean air and aerosol exposed cells. In the experiments we saw differences in toxicological responses both between the clean air and aerosol samples as well as between fresh and aged emissions.