10th International Aerosol Conference September 2 - September 7, 2018 America's Center Convention Complex St. Louis, Missouri, USA
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Application of Air-Liquid-Interface (ALI) Based In-Vitro Exposure of Human or Murine Lung Cells and Validation by Selected Animal Exposure Tests in the Framework of the HICE Consortium to Investigate Fresh and Aged Combustion Aerosols
RALF ZIMMERMANN, Maija-Riitta Hirvonen, Jorma Jokiniemi, Gunnar Dittmar, Jeroen Buters, Hanns Rudolf Paur, Carsten Weiß, Bert Buchholz, Tamara Kanashova, Sebastian Oeder, Marco Dilger, Tobias Krebs, Sven Ehlert, Thorsten Streibel, Juergen Schnelle-Kreis, Martin Sklorz, Stefanie Kasurinen, Sebastiano di Bucchianico, Johannes Passig, Jürgen Orasche, Mikko Happo, Hendryk Czech, Olli Sippula, Pasi Jalava, HICE Consortium, Helmholtz Zentrum München and Rostock University
Abstract Number: 1153 Working Group: Aerosol Exposure
Abstract Combustion aerosol emissions are responsible for acute and chronic health effects such as asthma exacerbation, heart-arrhythmia and heart-failure, lung cancer or COPD in humans. However, different combustion sources may have different health outcomes. Furthermore the effects may depend on the type of combustion compliance and can be mitigated/altered by abatement technologies. However, up to now only few links between aerosol chemical composition and biological effects have been established. Thus the relative toxicity of different combustion emissions needs to be determined experimentally. In the framework of the Virtual Helmholtz Institute-HICE (www.hice-vi.eu), physical and chemical properties of combustion emissions and their biological effects on lung cells (human epithelial cells: A549, BEAS2B and RAW/THP1 murine/human macrophages) are comprehensively analysed. For addressing the biological activity and toxicity of the aerosols, the lung cell-cultures were realistically exposed at the air-liquid interface (ALI) using 2 novel mobile automated ALI exposure-systems with 24 or 18 places for inserts with ca. 600.000 lung cells each (Vitrocell GmbH, Germany). The ALI systems are placed a mobile S2-bio safety laboratory (HICE MobiLab). One of the automated ALI systems is equipped with a newly developed exposure analysis system where the deposited mass, the particle size distribution (Topas GmbH, Germany) and the chemical composition is measured by mass spectrometry (Photonion GmbH). The cells are exposed with different moisturized com-bustion aerosols or clean air (reference). After 4h exposure biological effects were ana-lysed by multi-omics characterisation (transcriptomic, proteomic and metabolomics level) or assays for specific endpoints (viability, genotoxicity, cell integrity etc.). During all biological tests, the chemical composition and physical parameters of the emissions were thoroughly characterized (gases and particles). The acquired biological effects were comprehensively characterized and are put in context with the chemical and physical aerosol data (see e.g. Oeder et al., PLoSone 2015). For validation partly also animals (BL6 mice) are exposed in parallel to the ALI-exposures (RAW cells) and the macrophages from the broncheoalveolar lavage fluid (BALF) are subjected to molecular biological effects anaylsis as well. A first comparison of the effects from mice BALF (in-vivo) and RAW (in-vitro) for exposures with diluted exhaust from a Kubota diesel engine showed that immune response pathways are activated in both cases (in-vivo and in-vitro). Emissions from wood combustion (mansory heater, pellet burner and log wood stove, ship engines (heavy and light fuel oil), car engines (diesel, gasoline and ethanol fuels) were investigated by the ALI approach. Very recently lignite combustion and aging of wood and lignite aerosol emissions were investigated. The UV-light induced aging of the emission was performed in specially developed, high-throughput flow tube. In summary/conclusion observed biological response-strengths differ considerably for different combustion aerosol sources and are not well correlated to the deposited PM2.5-mass (i.e. partly also strong gas phase effects). This is suggesting large differences in the relative toxicity of the aerosol emissions from different combustion sources and fuel types. The aging experiments induce increased cytotoxicity, an effect which is currently deeper analyzed on the molecular biological level. In addition to adverse reactions also supposedly protective effects are observed. For example, the emission from a log-wood stove, exhibiting very high emission of soot, organics, PM2.5 are inducing only relatively mild acute effects in the ALI exposed cells, if compared to diesel or pellet burner emission. The high abundance of antioxidant compounds such polyphenols in the logwood stove-emissions may explain this counter-intuitive observation (Kanashova et al, J. Molec. Clin. Med., 2018). The latter findings are supported by detailed analyses of activated cellular response pathways (GO-term analysis), depicting regulation of pathways such as pro-inflammatory signaling, xenobiotic metabolism, phagocytosis or oxidative stress and findings from the selected animal exposure experiments.