Sandro Steiner

Diesel exhaust: current knowledge of adverse effects and underlying cellular mechanisms

Diesel engine emissions are among the most prevalent anthropogenic pollutants worldwide, and with the growing popularity of diesel-fueled engines in the private transportation sector, they are becoming increasingly widespread in densely populated urban regions. However, a large number of toxicological studies clearly show that diesel engine emissions profoundly affect human health. Thus the interest in the molecular and cellular mechanisms underlying these effects is large, especially concerning the nature of the components of diesel exhaust responsible for the effects and how they could be eliminated from the exhaust. This review describes the fundamental properties of diesel exhaust as well as the human respiratory tract and concludes that adverse health effects of diesel exhaust not only emerge from its chemical composition, but also from the interplay between its physical properties, the physiological and cellular properties, and function of the human respiratory tract. Furthermore, the primary molecular and cellular mechanisms triggered by diesel exhaust exposure, as well as the fundamentals of the methods for toxicological testing of diesel exhaust toxicity, are described. The key aspects of adverse effects induced by diesel exhaust exposure described herein will be important for regulators to support or ban certain technologies or to legitimate incentives for the development of promising new technologies such as catalytic diesel particle filters.

Cerium dioxide nanoparticles can interfere with the associated cellular mechanistic response to diesel exhaust exposure

The aim of this study was to compare the biological response of a sophisticated in vitro 3D co-culture model of the epithelial airway barrier to a co-exposure of CeO(2) NPs and diesel exhaust using a realistic air-liquid exposure system. Independent of the individual effects of either diesel exhaust or CeO(2) NPs investigation observed that a combined exposure of CeO(2) NPs and diesel exhaust did not cause a significant cytotoxic effect or alter cellular morphology after exposure to diesel exhaust for 2h at 20μg/ml (low dose) or for 6h at 60μg/ml (high dose), and a subsequent 6h exposure to an aerosolized solution of CeO(2) NPs at the same doses. A significant loss in the reduced intracellular glutathione level was recorded, although a significant increase in the oxidative marker HMOX-1 was found after exposure to a low and high dose respectively. Both the gene expression and protein release of tumour necrosis factor-α were significantly elevated after a high dose exposure only. In conclusion, CeO(2) NPs, in combination with diesel exhaust, can significantly interfere with the cell machinery, indicating a specific, potentially adverse role of CeO(2) NPs in regards to the biological response of diesel exhaust exposure.

Investigating the potential for different scooter and car exhaust emissions to cause cytotoxic and (pro-)inflammatory responses to a 3D in vitro model of the human epithelial airway

The aim of this study was to compare the cytotoxicity and the (pro-)inflammatory responses of two-stroke (direct injection and carburetor technology) and four-stroke scooter and diesel car exhaust emissions on lung cells in vitro. This was analyzed by exposing a 3D in vitro model of the epithelial airway (consisting of human bronchial epithelial cells (cell line 16HBE14o−) combined with human whole blood monocyte-derived macrophages and dendritic cells) to physically characterized exhaust emissions. Biological endpoints of cytotoxicity (lactate dehydrogenase release), as well as pro-inflammatory cytokine (tumor necrosis factor (TNF)-α) and inflammatory chemokine (interleukin(IL)-8) stimulation were examined. Two-stroke direct injection scooter exhaust contained the highest particle number concentration and nitrogen oxides (NO x ) concentrations and the emissions from the two-stroke carburetor scooter contained the highest hydrocarbon and lowest NO x concentrations. The four-stroke scooter emitted the highest carbon monoxide concentration whereas the cars emitted the lowest. The combination of various technical optimizations for the two-stroke direct injection scooter (particle filter, oxidative catalyst, better oil and fuel) reduced the total emissions strongly and the TNF-α concentration significantly (p < 0.05). The cytotoxicity and the IL-8 concentration showed strong tendencies to be reduced. The analysis of the emissions of all tested two-stroke, four-stroke scooters and diesel cars showed a strong association between the adverse effects and the particle number concentration.

Biological Effects in Lung Cells In Vitro of Exhaust Aerosols from a Gasoline Passenger Car With and Without Particle Filter

Exhaust aerosol from gasoline passenger cars is a complex mixture of a particulate fraction as well as volatile compounds. In contrary to the observed adverse effects of diesel exhaust particles the gasoline exhaust has, however, received little attention so far. The aim of this study was to perform a comparison of exhaust composition and biological responses from freshly produced non-filtered exhaust as well as from exhaust filtered with a noncoated gasoline particle filter (GPF). A 3D model of the human epithelial airway barrier was exposed to the exhaust directly at the air-liquid interface and different effects such as cytotoxicity, antioxidative response, pro-inflammation, and activation of the aryl hydrocarbon receptor (AhR) were studied. In addition, genotoxicity was assessed using the Ames test. By an online analysis of the exhaust, it has been shown that the GPF efficiently filters the particle count in both the cold and warm phase when the new European driving cycle (NEDC) was applied. The lung cell tests revealed that the use of the GPF increased the antioxidative glutathionine (GSH) response as well as the pro-inflammatory potential, i.e., IL-8, expression, indicating increased cell stimulation by the volatile compounds alone. The removal of the particulate fraction, however, decreased significantly the AhR activation in comparison to unfiltered exhaust, and the exhaust genotoxicity was reduced as tested by the Ames test. In conclusion, GPF exhaust did not completely reduce the adverse effects of gasoline exhaust in the in vitro test and further experiments with a coated GPF are needed in the future.

Test-methods on the test-bench: a comparison of complete exhaust and exhaust particle extracts for genotoxicity/mutagenicity assessment.

With the growing number of new exhaust after-treatment systems, fuels and fuel additives for internal combustion engines, efficient and reliable methods for detecting exhaust genotoxicity and mutagenicity are needed to avoid the widespread application of technologies with undesirable effects toward public health. In a commonly used approach, organic extracts of particulates rather than complete exhaust is used for genotoxicity/mutagenicity assessment, which may reduce the reliability of the results. In the present study, we assessed the mutagenicity and the genotoxicity of complete diesel exhaust compared to an organic exhaust particle extract from the same diesel exhaust in a bacterial and a eukaryotic system, that is, a complex human lung cell model. Both, complete exhaust and organic extract were found to act mutagenic/genotoxic, but the amplitudes of the effects differed considerably. Furthermore, our data indicate that the nature of the mutagenicity may not be identical for complete exhaust and particle extracts. Because in addition, differences between the responses of the different biological systems were found, we suggest that a comprehensive assessment of exhaust toxicity is preferably performed with complete exhaust and with biological systems representative for the organisms and organs of interest (i.e., human lungs) and not only with the Ames test.

Delivery efficiencies of constituents of combustion-derived aerosols across the air-liquid interface during in vitro exposures

In vitro aerosol exposure of epithelial cells grown at the air-liquid interface is an experimental methodology widely used in respiratory toxicology. The exposure depends to a large part on the physicochemical properties of individual aerosol constituents, as they determine the transfer kinetics from the aerosol into the cells. We characterized the transfer of 70 cigarette smoke constituents from the smoke into aqueous samples exposed in the Vitrocell® 24/48 aerosol exposure system. The amounts of these compounds in the applied smoke were determined by trapping whole smoke in N,N-dimethylformamide and then compared with their amounts in smoke-exposed, phosphate-buffered saline, yielding compound specific delivery efficiencies. Delivery efficiencies of different smoke constituents differed by up to five orders of magnitude, which indicates that the composition of the applied smoke is not necessarily representative for the delivered smoke. Therefore, dose metrics for in vitro exposure experiments should, if possible, be based on delivered and not applied doses. A comparison to literature on in vivo smoke retention in the respiratory tract indicated that the same applies for smoke retention in the respiratory tract.