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

Toxicol Appl Pharmacol. 2010 Dec 1;249(2):178-87. doi: 10.1016/j.taap.2010.09.005. Epub 2010 Sep 16.

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Biomarkers / metabolism
  • Blotting, Western
  • Cell Proliferation / drug effects
  • Dose-Response Relationship, Drug
  • Eicosanoids / biosynthesis
  • Histones / biosynthesis
  • Humans
  • Irritants / toxicity*
  • Keratinocytes / cytology
  • Keratinocytes / drug effects
  • Mustard Gas / analogs & derivatives*
  • Mustard Gas / toxicity
  • Poly Adenosine Diphosphate Ribose / biosynthesis
  • Proliferating Cell Nuclear Antigen / biosynthesis
  • RNA, Messenger / biosynthesis
  • Reverse Transcriptase Polymerase Chain Reaction
  • Skin / cytology
  • Skin / drug effects*
  • Time Factors

Substances

  • Biomarkers
  • Eicosanoids
  • H2AX protein, human
  • Histones
  • Irritants
  • Proliferating Cell Nuclear Antigen
  • RNA, Messenger
  • Poly Adenosine Diphosphate Ribose
  • 2-chloroethyl ethyl sulfide
  • Mustard Gas

Grant support