Adenovirus‐mediated iNOS gene transfer enhances recovery from skin injuries due to sulphur mustard exposure in vitro

Abstract

Sulfur mustard (SM) is a potent vesicating chemical warfare agent. One of the major specific target organs for SM is the skin, and injuries may take several months to heal and cause substantial functional and cosmetic deficits. There are strong correlations between reduced cutaneous nitric oxide (NO) synthesized by inducible NO synthase (iNOS) levels and impaired wound healing. We reported that SM downregulated iNOS expression to inhibit wound healing and that maintaining of NO level by iNOS gene transfer would be promising candidate therapeutics for SM skin injuries (Ishida et al., 2008). In this study, first we made adenovirus constructs using the AdenoVator(tm) system (Qbiogene) which encode the full length of the open frame of human iNOS cDNA (Ad‐iNOS). In vitro wound was created by scraping a pipetor (with 200‐l tips) in normal human epidermal keratinocytes (NHEK). SM (20 mM) exposure caused a significant reduction of iNOS expression with decreased NO production in NHEK. However, infection of NHEK with Ad‐iNOS 3 h before SM exposure resulted in complete recovery of iNOS expression and NO production. The inhibition of wound healing caused by SM was also recovered by the gene transfer. Mock infection did not show any recovery of iNOS expression, NO production, or wound healing after SM exposure. This work was supported by JSTO‐CBD/DTRA award # 2.T0002_05_WR_C