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EXPRESSION OF PROLIFERATIVE AND INFLAMMATORY MARKERS IN A FULL-THICKNESS HUMAN SKIN EQUIVALENT FOLLOWING EXPOSURE TO THE MODEL SULFUR MUSTARD VESICANT, 2-CHLOROETHYL ETHYL SULFIDE.

Black1, A.T., Hayden2, P.J., Casillas3, R.P., Heck4, D.E., Gerecke1, D.R., Sinko5, P.J., Laskin1, D.L., Laskin6, J.D. 1Pharmacology and Toxicology, Rutgers University, Piscataway, NJ, USA. 2MatTek Corporation, Ashland, MA, USA. 3Battelle Memorial Institute, Columbus, OH, USA. 4Environmental Health Sciences, New York Medical College, Valhalla, NY, USA. 5Pharmaceutics, Rutgers University, Piscataway, NJ, USA. 6Environmental and Occupational Medicine, UMDNJ-Robert Wood Johnson Medical School, Piscataway, NJ, USA.
Abstract

Sulfur mustard is a potent vesicant that induces inflammation, edema and blistering following dermal exposure. To assess molecular mechanisms mediating these responses, we analyzed the effects of the model sulfur mustard vesicant, 2-chloroethyl ethyl sulfide, on EpiDerm-FT™, a commercially available full-thickness human skin equivalent. CEES (100–1000 ìM) caused a concentration-dependent increase in pyknotic nuclei and vacuolization in basal keratinocytes; at high concentrations (300–1000 ìM), CEES also disrupted keratin filament architecture in the stratum corneum. This was associated with time-dependent increases in expression of proliferating cell nuclear antigen, a marker of cell proliferation, and poly(ADP-ribose) polymerase (PARP) and phosphorylated histone H2AX, markers of DNA damage. Concentration- and time- dependent increases in mRNA and protein expression of eicosanoid biosynthetic enzymes including COX-2, 5- lipoxygenase, microsomal PGE2 synthases, leukotriene (LT) A4 hydrolase and LTC4 synthase were observed in CEES-treated skin equivalents, as well as in antioxidant enzymes, glutathione S-transferases A1-2 (GSTA1-2), GSTA3 and GSTA4. These data demonstrate that CEES induces rapid cellular damage, cytotoxicity and inflammation in full-thickness skin equivalents. These effects are similar to human responses to vesicants in vivo and suggest that the full thickness skin equivalent is a useful in vitro model to characterize the biological effects of mustards and to develop potential therapeutics.

Keywords

5-LOX, Abnormal trichrome staining, COX-2.5-lipoxygenase, Cu,Zn-SOD, EFT-400, Eicosanoid biosynthetic enzymes, EpiDermFT™, GSTA3, GSTA4, Glutathione S-transferases A1-2 (GSTA1-2), Inflammation, LTC4 synthase, Leukotriene (LT) A4 hydrolase , Markers of inflammation, Microsomal PGE2 synthases, Mn-SOD, Morphologic changes, Nuclear condensation, Phosphorylated histone H2AX, Poly(ADP-ribose) polymerase (PARP), Proliferating cell nuclear antigen (PCNA), Prostaglandin biosysnthesis, Sulfur mustard, Vesicant

Materials Tested

2-chloroethyl ethyl sulfide

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