Nitric Oxide Synthase Gene Transfer Overcomes the Inhibition of Wound Healing by Sulfur Mustard in a Human Keratinocyte In Vitro Model

Hiroshi Ishida,Radharaman Ray,Jack Amnuaysirikul,Prabhati Ray,Keiko Ishida

doi:10.5402/2012/190429

Hiroshi Ishida, Radharaman Ray + Show 3 more

Open Access

https://doi.org/10.5402/2012/190429

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Journal: ISRN Toxicology	Publication Date: Nov 14, 2012
Citations: 23	License type: CC BY 3.0

Affiliation: Walter Reed Army Institute of Research

Abstract

Sulfur mustard (SM) is a chemical warfare agent that causes extensive skin injury. Previously we reported that SM exposure resulted in suppression of inducible nitric oxide synthase (iNOS) expression to inhibit the healing of scratch wounds in a cultured normal human epidermal keratinocyte (NHEK) model. Based on this finding, the present study was to use adenovirus-mediated gene transfer of iNOS to restore the nitric oxide (NO) supply depleted by exposure to SM and to evaluate the effect of NO on wound healing inhibited by SM in NHEKs. The effect of the iNOS gene transfer on iNOS protein expression and NO generation were monitored by Western blot and flow cytometry, respectively. Wound healing with or without the iNOS gene transfer after SM exposure was assessed by light and confocal microscopy. The iNOS gene transfer via adenovirus resulted in overexpression of the iNOS and an increase in NO production regardless of SM exposure in the NHEK model. The gene transfer was also effective in overcoming the inhibition of wound healing due to SM exposure leading to the promotion of wound closure. The findings in this study suggest that the iNOS gene transfer is a promising therapeutic strategy for SM-induced skin injury.

Highlights

Sulfur mustard (SM), bis-2-(chloroethyl) sulfide, is a chemical warfare agent that causes extensive skin injury
We tested the effect of the inducible nitric oxide synthase (iNOS) gene transfer by Ad-iNOS Infectious Viral Particles (AIVP) in normal human epidermal keratinocyte (NHEK) either without scratching or immediately after scratching on both the level of iNOS expression and nitric oxide (NO) production with or without SM treatment
We hypothesized that maintaining iNOS expression by iNOS gene transfer after SM exposure would be a good therapeutic for skin injuries

Summary

Introduction

Sulfur mustard (SM), bis-2-(chloroethyl) sulfide, is a chemical warfare agent that causes extensive skin injury. Frank et al [4] reported that the enhanced induction of vascular endothelial growth factor (VEGF) expression observed in keratinocytes after cytokine stimulation was dependent on the presence of endogenously produced NO during wound healing in vitro. NO modulates the level of certain chemoattractant cytokines such as interleukins and transforming growth factor (TGF) β1, which initiate post-wound inflammation, resulting in promotion of keratinocyte recruitment to wounds, proliferation, and differentiation [6]. These facts strongly suggest that NO plays an important role in skin-wound healing

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