title: Development of a repeated exposure protocol of human bronchial epithelium in vitro to study the long-term effects of atmospheric particles.
abstract: Chronic exposure to atmospheric particles is suspected of exacerbating chronic inflammatory respiratory diseases but the underlying mechanisms remain poorly understood. An experimental strategy using human bronchial epithelial cells (NHBE) known to be one of the main target cells of particles in the lung was developed to investigate the long term effects of repeated exposure to particles. Primary cultures of NHBE cells were grown at an air-liquid interface and subjected to repeated treatments to particles. Fate of particles, pro inflammatory response and epithelial differentiation were studied during the 5 weeks following the final treatment. Ultrastructural observations revealed the biopersistence of particles in the bronchial epithelium. The expression of cytochrome P450 1A1, was transiently induced, suggesting that organic compounds could have been metabolized. The release of GM-CSF and IL-6 (biomarkers of pro-inflammatory response), was induced by particle treatments and was maintained up to 5weeks after treatments. The release of amphiregulin and TGFα (Growth Factor) was induced after each treatment. The number of cells expressing the mucin MUC5AC, a differentiation marker, was increased in particle-exposed epithelium. The experimental strategy we developed is suitable for investigating in greater depth the long term effects of particles on bronchial epithelial cells repeatedly exposed to atmospheric particles in vitro.
authors.affiliation: Univ Paris Diderot, Sorbonne Paris Cité, Laboratory of Molecular and Cellular Responses to Xenobiotics, Unit of Functional and Adaptive Biology (BFA) EAC CNRS 4413, 75 013 Paris, France. boublil.laura@gmail.com
authors.forename: Laura
authors.initials: L
authors.lastname: Boublil
authors.forename: Emeline
authors.initials: E
authors.lastname: Assémat
authors.forename: Marie-Caroline
authors.initials: MC
authors.lastname: Borot
authors.forename: Sonja
authors.initials: S
authors.lastname: Boland
authors.forename: Laurent
authors.initials: L
authors.lastname: Martinon
authors.forename: Jean
authors.initials: J
authors.lastname: Sciare
authors.forename: Armelle
authors.initials: A
authors.lastname: Baeza-Squiban
mesh_terms: D000393:Air Pollutants / Q000633:toxicity*; D001980:Bronchi; D045744:Cell Line, Tumor; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics; D016207:Cytokines / Q000378:metabolism; D006801:Humans; D046529:Microscopy, Electron, Transmission; D052638:Particulate Matter / Q000008:administration & dosage / Q000633:toxicity*; D012333:RNA, Messenger / Q000378:metabolism; D020545:Respiratory Mucosa / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D000393:Air Pollutants; D016207:Cytokines; D052638:Particulate Matter; D012333:RNA, Messenger; C569463:CYP1A1 protein, human; D019363:Cytochrome P-450 CYP1A1