title: 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.
authors.affiliation: Pharmacology and Toxicology, Rutgers University, Piscataway, NJ 08854, USA.
authors.forename: Adrienne T
authors.initials: AT
authors.lastname: Black
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Hayden
authors.forename: Robert P
authors.initials: RP
authors.lastname: Casillas
authors.forename: Diane E
authors.initials: DE
authors.lastname: Heck
authors.forename: Donald R
authors.initials: DR
authors.lastname: Gerecke
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Sinko
authors.forename: Debra L
authors.initials: DL
authors.lastname: Laskin
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Laskin
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D015777:Eicosanoids / Q000096:biosynthesis; D006657:Histones / Q000096:biosynthesis; D006801:Humans; D007509:Irritants / Q000633:toxicity*; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects; D009151:Mustard Gas / Q000031:analogs & derivatives* / Q000633:toxicity; D011064:Poly Adenosine Diphosphate Ribose / Q000096:biosynthesis; D018809:Proliferating Cell Nuclear Antigen / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000166:cytology / Q000187:drug effects*; D013997:Time Factors
chemical_list: D015415:Biomarkers; D015777:Eicosanoids; C468783:H2AX protein, human; D006657:Histones; D007509:Irritants; D018809:Proliferating Cell Nuclear Antigen; D012333:RNA, Messenger; D011064:Poly Adenosine Diphosphate Ribose; C031278:2-chloroethyl ethyl sulfide; D009151:Mustard Gas