Acute cytotoxicity and increased vascular endothelial growth factor after in vitro nitrogen mustard vapor exposure

RATIONALE:  Nitrogen mustard (NM) is a highly toxic alkylating agent.  It is classified as chemical threat due to its potential use as a warfare agent.  When inhaled, mustard exposure can cause both acute and chronic lung injury.  The aims of this study are to develop an in vitro culture system for modeling mustard-induced airway injury and to identify growth factors contributing to airway pathology.

METHODS:  Primary human bronchial epithelial cells co-cultured with primary human pulmonary endothelial cells were exposed to NM (25, 50, 100, 250 or 500 µM) or PBS (control) for 1 hour.  Lactate dehydrogenase (LDH) and trans-epithelial electrical resistance (TEER) were measured prior to and 24 hours after NM exposure.  Endothelial cultures were then stained for live/dead staining.  Exposed airway media was also analyzed for growth factors using Luminex multiplex technology. 

RESULTS:  A one-hour NM vapor exposure increased supernatant LDH and decreased TEER at 24 hours for all cu...