Anton Wessels

Oxidant generation and toxicity of size-fractionated ambient particles in human lung epithelial cells.

Exposure to ambient particulate matter (PM) is associated with respiratory and cardiovascular disease and lung cancer. In this study, we used size fractionated PM samples (3-7, 1.5-3, 0.95-1.5, 0.5-0.95, and <0.5 microm), collected at four contrasting locations (three urban sites, one remote background) in the UK with a Sierra-Andersen high volume cascade impactor. The H(2)O(2)-dependent oxidant generating capacity of the samples was determined by electron spin resonance with 5,5-dimethyl-1-pyrroline-N-oxide spin trapping. In A549 human lung epithelial cells, we determined the cytotoxicity of samples by LDH assay, and interleukin-8 (IL-8) release as an indicator of their inflammatory potency. Oxidative DNA damage was measured by the formamido-pyrimidine-glycosylase (fpg)-modified comet assay. Marked contrasts were observed for all endpoints. Remote background PM showed the lowest oxidant potential, was neither cytotoxic nor genotoxic and did not increase IL-8 release. For the other samples, effects were found to depend more on sampling location than on size fraction. PM collected at high-traffic locations generally showed the strongest oxidant capacity and toxicity. Significant correlations were observed between the oxidant generating potential and all toxicological endpoints investigated, which demonstrates that measurement of the oxidant generating potential by ESR represents a sensitive method to estimate the toxic potential of PM.

Neutrophil-derived ROS contribute to oxidative DNA damage induction by quartz particles

The carcinogenicity of respirable quartz is considered to be driven by reactive oxygen species (ROS) generation in association with chronic inflammation. The contribution of phagocyte-derived ROS to inflammation, oxidative stress, and DNA damage responses was investigated in the lungs of C57BL/6J wild-type and p47(phox-/-) mice, 24h after pharyngeal aspiration of DQ12 quartz (100 mg/kg bw). Bone-marrow-derived neutrophils from wild-type and p47(phox-/-) mice were used for parallel in vitro investigations in coculture with A549 human alveolar epithelial cells. Quartz induced a marked neutrophil influx in both wild-type and p47(phox-/-) mouse lungs. Significant increases in mRNA expression of the oxidative stress markers HO-1 and γ-GCS were observed only in quartz-treated wild-type animals. Oxidative DNA damage in lung tissue was not affected by quartz exposure and did not differ between p47(phox-/-) and WT mice. Differences in mRNA expression of the DNA repair genes OGG1, APE-1, DNA Polβ, and XRCC1 were also absent. Quartz treatment of cocultures containing wild-type neutrophils, but not p47(phox-/-) neutrophils, caused increased oxidative DNA damage in epithelial cells. Our study demonstrates that neutrophil-derived ROS significantly contribute to pulmonary oxidative stress responses after acute quartz exposure, yet their role in the associated induction of oxidative DNA damage could be shown only in vitro.

Oxidative stress and DNA damage responses in rat and mouse lung to inhaled carbon nanoparticles

Abstract We have investigated whether short-term nose-only inhalation exposure to electric spark discharge-generated carbon nanoparticles (∼60 nm) causes oxidative stress and DNA damage responses in the lungs of rats (152 μg/m3; 4 h) and mice (142 μg/m3; 4 h, or three times 4 h). In both species, no pulmonary inflammation and toxicity were detected by bronchoalveolar lavage or mRNA expression analyses. Oxidative DNA damage (measured by fpg-comet assay), was also not increased in mouse whole lung tissue or isolated lung epithelial cells from rat. In addition, the mRNA expressions of the DNA base excision repair genes OGG1, DNA Polβ and XRCC1 were not altered. However, in the lung epithelial cells isolated from the nanoparticle-exposed rats a small but significant increase in APE-1 mRNA expression was measured. Thus, short-term inhalation of carbon nanoparticles under the applied exposure regimen, does not cause oxidative stress and DNA damage in the lungs of healthy mice and rats.

Investigation of the effects of short-term inhalation of carbon nanoparticles on brains and lungs of c57bl/6j and p47phox−/− mice

Recent studies indicate that the brain is a target for toxic carbonaceous nanoparticles present in ambient air. It has been proposed that the neurotoxic effects of such particles are driven by nicotinamide adenine dinucleotide phosphate (NADPH) oxidase mediated generation of reactive oxygen species (ROS) in activated microglia. In the present study, we have evaluated the effects of short term (4h) nose-only inhalation exposure to carbon NP (CNP) in the brains and lungs of C57BL/6J mice and in p47(phox-/-) mice that lack a functional NADPH oxidase. It was shown that the lungs of the p47(phox-/-) mice are less responsive to CNP inhalation than lungs of the corresponding C57BL/6J control animals. Lung tissue mRNA expression of the oxidative stress/DNA damage response genes 8-oxoguanine glycosylase (OGG1) and apurinic/apyrimidinic endonuclease 1 (APE1) were induced by CNP exposure in C57BL/6J but not in the p47(phox-/-) mice. In contrast, the expression of these genes, as well as Tumor Necrosis Factor-α (TNFα), Cyclooxygenase-2 (COX-2) and Heme Oxygenase-1 (HO-1) was not altered in the olfactory bulb, cerebellum or remaining brain tissue part of either mouse background. This indicates that neuroinflammation was not induced by this exposure. CNP inhalation for 4h or for 4h on three consecutive days also did not affect brain tissue protein expression of interleukin (IL)-1β, while a clear significant difference in constitutive expression level of this pro-inflammatory cytokine was found between C57BL/6J and p47(phox-/-) mice. In conclusion, short-term inhalation exposure to pure carbon nanoparticles can trigger mild p47(phox) dependent oxidative stress responses in the lungs of mice whereas in their brains at the same exposure levels signs of oxidative stress and inflammation remain absent. The possible role of p47(phox) in the neuro-inflammatory effects of nanoparticles in vivo remains to be clarified.