title: Effects of acute in vitro exposure of murine precision-cut lung slices to gaseous nitrogen dioxide and ozone in an air-liquid interface (ALI) culture.
abstract: The aim of this study was to establish an air-liquid interface (ALI) culture of precision-cut lung slices (PCLS) for direct exposure of lung cells to gaseous contaminants. Nitrogen dioxide (NO(2)) and ozone (O(3)) were selected as model gas compounds. Acute pro-inflammatory and toxic effects of NO(2) and O(3) on live lung tissue were investigated. Murine PCLS were exposed to different flow rates (3-30mL/min) of synthetic air, O(3) (3.5-8.5ppm), or NO(2) (1-80ppm). Tissue survived ex vivo in ALI culture and resisted exposure to NO(2) (1-10ppm) and O(3) (3.5-8.5ppm) for 1h. Longer exposure to NO(2) resulted in a clear loss of viability, whereas exposure to O(3) was less effective. Exposure to NO(2) dose-dependently induced release of the pro-inflammatory IL-1alpha (40%), whereas RANTES, IL-12, and eotaxin remained unchanged. Early secretion of IL-1alpha (80%), RANTES (>800%), MIP-1beta (44%), and MCP-1 (60%) was already detected after 1h of exposure to O(3). The obtained data showed that direct exposure to O(3) and NO(2) induced cytotoxicity and pro-inflammatory responses in PCLS with ALI culture. This provides a model that more closely resembles in vivo exposure of airborne contaminants, and thus should be appropriate for toxicity testing.
authors.affiliation: Fraunhofer Institute for Toxicology and Experimental Medicine, Nikolai-Fuchs-Str. 1, 30625 Hannover, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Switalla
authors.forename: J
authors.initials: J
authors.lastname: Knebel
authors.forename: D
authors.initials: D
authors.lastname: Ritter
authors.forename: N
authors.initials: N
authors.lastname: Krug
authors.forename: A
authors.initials: A
authors.lastname: Braun
authors.forename: K
authors.initials: K
authors.lastname: Sewald
mesh_terms: D000818:Animals; D018932:Chemokine CCL2 / Q000378:metabolism; D054407:Chemokine CCL4 / Q000378:metabolism; D018946:Chemokine CCL5 / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005260:Female; D005740:Gases; D018836:Inflammation Mediators / Q000378:metabolism; D018664:Interleukin-12 / Q000378:metabolism; D053582:Interleukin-1alpha / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008168:Lung / Q000187:drug effects* / Q000276:immunology / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D009585:Nitrogen Dioxide / Q000633:toxicity*; D010126:Ozone / Q000633:toxicity*; D013997:Time Factors; D046509:Tissue Culture Techniques; D014022:Tissue Survival / Q000187:drug effects
chemical_list: C491768:Ccl2 protein, mouse; C496535:Ccl4 protein, mouse; C503460:Ccl5 protein, mouse; D018932:Chemokine CCL2; D054407:Chemokine CCL4; D018946:Chemokine CCL5; D005740:Gases; D018836:Inflammation Mediators; D053582:Interleukin-1alpha; D008070:Lipopolysaccharides; C440499:lipopolysaccharide, Escherichia coli O111 B4; D018664:Interleukin-12; D010126:Ozone; D009585:Nitrogen Dioxide