Assessment of biological responses of EpiAirway 3-D cell constructs versus A549 cells for determining toxicity of ambient air pollution

Abstract Context: EpiAirwayTM 3-D constructs are human-derived cell cultures of differentiated airway epithelial cells that may represent a more biologically relevant model of the human lung. However, limited information is available on their utility for exposures to air pollutants at the air-liquid interface (ALI). Objective: To assess the biological responses of EpiAirwayTM cells in comparison to the responses of A549 human alveolar epithelial cells after exposure to air pollutants at ALI. Methods: Cells were exposed to filtered air, 400 ppb of ozone (O3) or a photochemically aged Synthetic Urban Mixture (SynUrb54) consisting of hydrocarbons, nitrogen oxides, O3 and other secondary oxidation products for 4 h. Basolateral supernatants and apical washes were collected at 9 and 24 h post-exposure. We assessed cytotoxicity by measuring lactate dehydrogenase (LDH) release into the culture medium and apical surface. Interleukin 6 (IL-6) and interleukin 8 (IL-8) proteins were measured in the culture medium and in the apical washes to determine the inflammatory response after exposure. Results: Both O3 and SynUrb54 significantly increased basolateral levels of LDH and IL-8 in A549 cells. No significant changes in LDH and IL-8 levels were observed in the EpiAirwayTM cells, however, IL-6 in the apical surface was significantly elevated at 24 h after O3 exposure. Conclusion: LDH and IL-8 are robust endpoints for assessing toxicity in A549 cells. The EpiAirwayTM cells show minimal adverse effects after exposure suggesting that they are more toxicologically resistant compared to A549 cells. Higher concentrations or longer exposure times are needed to induce effects on EpiAirwayTM cells.

[1]  Steven K. Gibb.  Toxicity testing in the 21st century: a vision and a strategy. , 2008, Reproductive toxicology.

[2]  Gerhard Kasper,et al.  In-vitro cell exposure studies for the assessment of nanoparticle toxicity in the lung - A dialog between aerosol science and biology , 2011 .

[3]  Dominique Balharry,et al.  Proteomic profiling of human respiratory epithelia by iTRAQ reveals biomarkers of exposure and harm by tobacco smoke components , 2011, Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals.

[4]  Ilona Jaspers,et al.  Photochemical Products in Urban Mixtures Enhance Inflammatory Responses in Lung Cells , 2004, Inhalation toxicology.

[5]  Gary R. Grotendorst,et al.  Sulfur mustard vapor effects on differentiated human lung cells , 2010, Inhalation toxicology.

[6]  I. Jaspers,et al.  The Gillings Sampler--an electrostatic air sampler as an alternative method for aerosol in vitro exposure studies. , 2014, Chemico-biological interactions.

[7]  J. Wong,et al.  Polarized Secretion of Interleukin (IL)-6 and IL-8 by Human Airway Epithelia 16HBE14o- Cells in Response to Cationic Polypeptide Challenge , 2010, PloS one.

[8]  J. Mauderly,et al.  RESPONSES OF DIFFERENTIATED PRIMARY HUMAN LUNG EPITHELIAL CELLS TO EXPOSURE TO DIESEL EXHAUST AT AN AIR-LIQUID INTERFACE , 2007, Experimental lung research.

[9]  S. Khurshid,et al.  Toxicological analysis of limonene reaction products using an in vitro exposure system. , 2013, Toxicology in vitro : an international journal published in association with BIBRA.

[10]  I. Jaspers,et al.  Gaseous VOCs rapidly modify particulate matter and its biological effects – Part 2 : Complex urban VOCs and model PM , 2012 .

[11]  Manuel C. Peitsch,et al.  Systems Approaches Evaluating the Perturbation of Xenobiotic Metabolism in Response to Cigarette Smoke Exposure in Nasal and Bronchial Tissues , 2013, BioMed research international.

[12]  Susanne Krauss-Etschmann,et al.  Health effects of ambient particulate matter--biological mechanisms and inflammatory responses to in vitro and in vivo particle exposures. , 2008, Inhalation toxicology.

[13]  D. Morgan,et al.  Diacetyl induces amphiregulin shedding in pulmonary epithelial cells and in experimental bronchiolitis obliterans. , 2014, American journal of respiratory cell and molecular biology.

[14]  Kelly Bérubé,et al.  Human primary bronchial lung cell constructs: the new respiratory models. , 2010, Toxicology.

[15]  R. Panettieri,et al.  Ozone modulates IL-6 secretion in human airway epithelial and smooth muscle cells. , 2009, American journal of physiology. Lung cellular and molecular physiology.

[16]  Barbara Rothen-Rutishauser,et al.  A dose-controlled system for air-liquid interface cell exposure and application to zinc oxide nanoparticles , 2009, Particle and Fibre Toxicology.

[17]  C. Roth,et al.  Deposition of fine and ultrafine aerosol particles during exposure at the air/cell interface , 2002 .

[18]  S. Randell,et al.  Novel human bronchial epithelial cell lines for cystic fibrosis research. , 2009, American journal of physiology. Lung cellular and molecular physiology.

[19]  David Diaz-Sanchez,et al.  Ozone induces a proinflammatory response in primary human bronchial epithelial cells through mitogen-activated protein kinase activation without nuclear factor-κB activation. , 2014, American journal of respiratory cell and molecular biology.

[20]  G. Evans,et al.  In vivo and In vitro Assessment of Particulate Matter Toxicology , 2010 .

[21]  I. Jaspers,et al.  Effects of 1,3-Butadiene, Isoprene, and Their Photochemical Degradation Products on Human Lung Cells , 2004, Environmental health perspectives.

[22]  U. Mohr,et al.  A modified CULTEX system for the direct exposure of bacteria to inhalable substances. , 2004, Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie.

[23]  I. Jaspers,et al.  Atmospheric photochemical transformations enhance 1,3-butadiene-induced inflammatory responses in human epithelial cells: The role of ozone and other photochemical degradation products. , 2007, Chemico-biological interactions.

[24]  R. Fry,et al.  A Toxicogenomic Comparison of Primary and Photochemically Altered Air Pollutant Mixtures , 2011, Environmental health perspectives.

[25]  M. McKimJames.  The Journal of Applied In Vitro Toxicology: Its Time Has Come! , 2015 .

[26]  D. Daines,et al.  Use of the EpiAirway model for characterizing long-term host-pathogen interactions. , 2011, Journal of visualized experiments : JoVE.

[27]  D. Hochrainer,et al.  The CULTEX RFS: A Comprehensive Technical Approach for the In Vitro Exposure of Airway Epithelial Cells to the Particulate Matter at the Air-Liquid Interface , 2013, BioMed research international.

[28]  J. Volckens,et al.  Direct particle-to-cell deposition of coarse ambient particulate matter increases the production of inflammatory mediators from cultured human airway epithelial cells. , 2009, Environmental science & technology.

[29]  M. Peitsch,et al.  Human bronchial epithelial cells exposed in vitro to cigarette smoke at the air-liquid interface resemble bronchial epithelium from human smokers. , 2013, American journal of physiology. Lung cellular and molecular physiology.

[30]  M. Strand,et al.  SERCA2 Regulates Non-CF and CF Airway Epithelial Cell Response to Ozone , 2011, PloS one.

[31]  M. J. Charles,et al.  A sensitive method for the quantification of acrolein and other volatile carbonyls in ambient air. , 2006, Analytical chemistry.

[32]  A. Hickey,et al.  Cellular response to the deposition of diesel exhaust particle aerosols onto human lung cells grown at the air-liquid interface by inertial impaction. , 2011, Toxicology in vitro : an international journal published in association with BIBRA.

[33]  J. Rička,et al.  A novel exposure system for the efficient and controlled deposition of aerosol particles onto cell cultures. , 2008, Environmental science & technology.

[34]  Z. Ning,et al.  Field evaluation of a new particle concentrator- electrostatic precipitator system for measuring chemical and toxicological properties of particulate matter , 2008, Particle and Fibre Toxicology.

[35]  J. Curtis,et al.  Increased cytokine response of rhinovirus-infected airway epithelial cells in chronic obstructive pulmonary disease. , 2010, American journal of respiratory and critical care medicine.

[36]  H. Burtscher,et al.  Acute toxicity of silver and carbon nanoaerosols to normal and cystic fibrosis human bronchial epithelial cells , 2016, Nanotoxicology.

[37]  U. Baltensperger,et al.  Responses of lung cells to realistic exposure of primary and aged carbonaceous aerosols , 2013 .

[38]  S. Randell,et al.  Cytokine secretion by cystic fibrosis airway epithelial cells. , 2004, American journal of respiratory and critical care medicine.

[39]  W. Cascio,et al.  Photochemically Altered Air Pollution Mixtures and Contractile Parameters in Isolated Murine Hearts before and after Ischemia , 2013, Environmental health perspectives.

[40]  Christian Mühlfeld,et al.  In vitro models of the human epithelial airway barrier to study the toxic potential of particulate matter. , 2008, Expert opinion on drug metabolism & toxicology.

[41]  I. Jaspers,et al.  In vitro exposures in diesel exhaust atmospheres: resuspension of PM from filters versus direct deposition of PM from air. , 2012, Environmental science & technology.

[42]  U. Mohr,et al.  CULTEX--a new system and technique for the cultivation and exposure of cells at the air/liquid interface. , 1999, Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie.

[43]  J. Bailar,et al.  Toxicity Testing in the 21st Century: A Vision and a Strategy , 2010, Journal of toxicology and environmental health. Part B, Critical reviews.

[44]  David Diaz-Sanchez,et al.  Progress in assessing air pollutant risks from in vitro exposures: matching ozone dose and effect in human airway cells. , 2014, Toxicological sciences : an official journal of the Society of Toxicology.

[45]  K. BéruBé,et al.  An in vitro approach to assess the toxicity of inhaled tobacco smoke components: nicotine, cadmium, formaldehyde and urethane. , 2008, Toxicology.

[46]  B. Rothen‐Rutishauser,et al.  An optimized in vitro model of the respiratory tract wall to study particle cell interactions. , 2006, Journal of aerosol medicine : the official journal of the International Society for Aerosols in Medicine.