Alveolar epithelial cells (A549) exposed at the air–liquid interface to diesel exhaust: First study in TNO’s powertrain test center

Kooter, Ingeborg M.; Alblas, M.J.; Jedyńska, Aleksandra; Steenhof, Maaike; Houtzager, Marc M.G.; Ras, Martijn van

doi:10.1016/j.tiv.2013.10.007

Cited by 42 publications

(23 citation statements)

References 42 publications

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“…Exposure of submerged cells to sampled exhaust components suspended in cell culture medium does not represent a realistic scenario; therefore, many sophisticated systems have been developed recently to bring a freshly produced exhaust directly onto the lung cells [21,[25][26][27][28][29]. Exhaust samples can be taken directly at the tailpipe and brought to the lung cell cultures with a delay of less than 30 s [29].…”

Section: Electronic Supplementary Materialsmentioning

confidence: 99%

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

Bisig

Steiner

Comte

et al. 2015

Emiss. Control Sci. Technol.

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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 proinflammatory 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.

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Section: Electronic Supplementary Materialsmentioning

confidence: 99%

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

Bisig

Steiner

Comte

et al. 2015

Emiss. Control Sci. Technol.

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“…19,20 However, a decrease in IL-8 was also demonstrated by Kooter et al, 21 in the presence of DEA-induced oxidative stress and cytotoxicity, with 1.5 h of active exposure and 24 h of postexposure. Considering the short-term exposure, it cannot be excluded that pro-inflammatory responses may have occurred at later time points or even in a temperaturized chamber.…”

Section: Environmental Science and Technologymentioning

confidence: 73%

Enhanced Deposition by Electrostatic Field-Assistance Aggravating Diesel Exhaust Aerosol Toxicity for Human Lung Cells

Stoehr

Madl

Boyles

et al. 2015

Environ. Sci. Technol.

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Air pollution is associated with increased risk of cardiovascular and pulmonary diseases, but conventional air quality monitoring gives no information about biological consequences. Exposing human lung cells at the air-liquid interface (ALI) to ambient aerosol could help identify acute biological responses. This study investigated electrode-assisted deposition of diesel exhaust aerosol (DEA) on human lung epithelial cells (A549) in a prototype exposure chamber. A549 cells were exposed to DEA at the ALI and under submerged conditions in different electrostatic fields (EFs) and were assessed for cell viability, membrane integrity, and IL-8 secretion. Qualitative differences of the DEA and its deposition under different EFs were characterized using scanning mobility particle sizer (SMPS) measurements, transmission electron microscopy (TEM), and electron energy loss spectroscopy (EELS). Upon exposure to DEA only, cell viability decreased and membrane impairment increased for cells at the ALI; submerged cells were unaffected. These responses were enhanced upon application of an EF, as was DEA deposition. No adverse effects were observed for filtered DEA or air only, confirming particle-induced responses. The prototype exposure chamber proved suitable for testing DEA-induced biological responses of cells at the ALI using electrode-assisted deposition and may be useful for analysis of other air pollutants.

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“…Although an extensive discussion of the various read-outs is beyond the scope of this review, it needs to be stressed that a range of other parameters are currently assessed to determine the biological effect of an inhaled toxicant in in vitro cell culture systems. These include the various -omics techniques, as illustrated by the effect of electronic cigarette liquid on the metabolome of airway epithelial cells (Aug et al, 2015), markers of oxidative stress, induction of an oxidative stress response and inflammatory response (Zarcone et al, 2016;Zarcone et al, 2017;Kuper et al, 2015;Kooter et al, 2013), induction of the integrated stress response (Zarcone et al, 2016;Zarcone et al, 2017), impairment of autophagy (Shivalingappa et al, 2015), and impairment of epithelial antimicrobial defence (Amatngalim et al, 2017;Herr et al, 2009;Wu et al, 2014). Adsorption of test compounds can be specifically addressed by using radiolabeled substances that are applied via the apical surface, as demonstrated by Reus et al using the MucilAirTM model of differentiated human bronchial epithelial cells (Reus et al, 2014).…”

Section: Mass Flow Controllermentioning

confidence: 99%

Human lung epithelial cell cultures for analysis of inhaled toxicants: Lessons learned and future directions

Hiemstra

Grootaers

Does

et al. 2018

Toxicology in Vitro

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153

116

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A B S T R A C TThe epithelium that covers the conducting airways and alveoli is a primary target for inhaled toxic substances, and therefore a focus in inhalation toxicology. The increasing concern about the use of animal models has stimulated the development of in vitro cell culture models for analysis of the biological effects of inhaled toxicants. However, the validity of the current in vitro models and their acceptance by regulatory authorities as an alternative to animal models is a reason for concern, and requires a critical review. In this review, focused on human lung epithelial cell cultures as a model for inhalation toxicology, we discuss the choice of cells for these models, the cell culture system used, the method of exposure as well as the various read-outs to assess the cellular response. We argue that rapid developments in the 3D culture of primary epithelial cells, the use of induced pluripotent stem cells for generation of lung epithelial cells and the development of organ-on-a-chip technology are among the important developments that will allow significant advances in this field. Furthermore, we discuss the various routes of application of inhaled toxicants by air-liquid interface models as well as the vast array of read-outs that may provide essential information. We conclude that close collaboration between researchers from various disciplines is essential for development of valid methods that are suitable for replacement of animal studies for inhalation toxicology.

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Alveolar epithelial cells (A549) exposed at the air–liquid interface to diesel exhaust: First study in TNO’s powertrain test center

Cited by 42 publications

References 42 publications

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

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

Enhanced Deposition by Electrostatic Field-Assistance Aggravating Diesel Exhaust Aerosol Toxicity for Human Lung Cells

Human lung epithelial cell cultures for analysis of inhaled toxicants: Lessons learned and future directions

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