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

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 μM) caused a concentration-dependent increase in pyknotic nuclei and vacuolization in basal keratinocytes; at high concentrations (300–1000 μM), 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.

Mitigation of nitrogen mustard mediated skin injury by a novel indomethacin bifunctional prodrug

“…NM exposure resulted in upregulation of iNOS and COX-2 in basal and superbasal cells after 1–3 days. Increases in COX-2 expression are consistent with previous findings that sulfur mustard, and the related half mustard, 2-chloroethyl ethyl sulfide, stimulate the release of prostaglandins from human and mouse keratinocytes, and that in mouse skin, sulfur mustard-induced injury is associated with increased COX-2 (Black et al, 2010; Joseph et al, 2011; Lefkowitz and Smith, 2002). While sebocytes continued to express iNOS and COX-2 for 1 day post NM, subsequently they degenerated.…”

Avian Collagen Is Useful for the Construction of Skin Equivalents

“…In SEs, PCNA is usually expressed in the basal layer [Black et al, 2010]. Consistent with this, PCNA was mainly detected in the basal layer of the control model.…”

Structural changes in hair follicles and sebaceous glands of hairless mice following exposure to sulfur mustard

“…Previous studies have shown that SM induces DNA double strand breaks, a process that activates DNA repair (Black et al, 2010; Jowsey et al, 2010). Histone phospho-H2A.X binds to double stranded DNA breaks creating a repair recognition domain that is important in controlling DNA damage (Clingen et al , 2008; Mah et al , 2010).…”

“…We have previously demonstrated that SM treatment of mouse epidermis causes rapid (within 1 day) increases in expression of phospho-H2A.X in the nucleus of basal keratinocytes (Joseph et al , 2011). The half mustard, 2-chloroethylethylamine, also induces phospho-H2A.X in basal keratinocytes of human full-skin constructs (Black et al, 2010). The present studies demonstrate that SM also induces double strand DNA breaks in hair follicles and sebaceous glands within 1 day of treatment.…”