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.
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.
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
2-chloroethyl ethyl sulfide
Request a copy of this paper, click here.