ToxSci Advance Access originally published online on September 11, 2009
Toxicological Sciences 2009 112(2):482-489; doi:10.1093/toxsci/kfp211
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Acute Morphological and Toxicological Effects in a Human Bronchial Coculture Model after Sulfur Mustard Exposure
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* Institute of Pathology, Johannes Gutenberg University Mainz, Mainz, Germany
Bundeswehr Institute of Pharmacology and Toxicology Munich, Munich, Germany
Clinic for Thorax Surgery, Catholic Clinical Centre St. Hildegardis, Mainz, Germany
1 To whom correspondence should be addressed. Fax: +49 (0) 6131 17 5645. E-mail: pohlc{at}uni-mainz.de.
Received July 8, 2009; accepted September 1, 2009
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Abstract |
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Sulfur mustard (SM) is a strong alkylating agent. Inhalation of SM causes acute lung injury accompanied by severe disruption of the airway barrier. In our study, we tested the acute effects after mustard exposure in an in vitro coculture bronchial model of the proximal barrier. To achieve this, we seeded normal human bronchial epithelial explant-outgrowth cells (HBEC) together with lung fibroblasts as a bilayer on filter plates and exposed the bronchial model after 31 days of differentiation to various concentrations of SM (30, 100, 300, and 500µM). The HBEC formed confluent layers, expressing functional tight junctions as measured by transepithelial electrical resistance (TER). Mucus production and cilia formation reappeared in the coculture model. TER was measured after 2 and 24 h following treatment. Depending on the different concentrations, TER decreased in the first 2 h up to 55% of the control at the highest concentration. After 24 h, TER seemed to recover because at concentrations up to 300µM values were equal to the control. SM induced a widening of intercellular spaces and a loss in cell-matrix adhesion. Mucus production increased with the result that cilia ceased to beat. Changes in the proinflammatory cytokines interleukin (IL)-6 and IL-8 were also observed. Apoptotic markers such as cytochrome c, p53, Fas-associated protein with death domain, and procaspase-3 were significantly induced at concentrations of less than 100µM. In summary, SM induces morphological and biochemical changes that reflect pathological effects of SM injury in vivo. It is hoped to use this coculture model to understand further the pathogenesis of SM-induced barrier injury and to search for novel approaches in SM therapy.
Key Words: primary bronchial cells; coculture; sulfur mustard; lung; barrier.
2 These authors contributed equally to this study.