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Ultrastructural characterization of sulfur mustard‐induced vesication in isolated perfused porcine skin

The isolated perfused porcine skin flap (IPPSF) is a novel alternative, humane in vitro model consisting of a viable epidermis and dermis with a functional microvasculature. For this study, 200 μl of either 10.0, 5.0, 2.5, 1.25, 0.50, or 0.20 mg/ml of bis (2‐chloroethyl) sulfide (HD) in ethanol or ethanol control was topically applied to a 5.0 cm2 dosing area of the IPPSF and perfused for 8 h with recirculating media. HD dermatotoxicity was assessed in the flap by cumulative glucose utilization (CGU), vascular resistance (VR), light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). HD produced a statistically significant dose relationship for gross blisters and microvesicles. The HD‐treated IPPSFs were also characterized by a decrease in CGU and an increase in VR. Light microscopic changes included mild intracellular and slight intercellular epidermal edema, multifocal epidermal‐dermal separation, and dark basal cells. Ultrastructural alterations consisted of cytoplasmic vacuoles, pyknotic basal cells, nucleolar segregation, and epidermal‐dermal separation occurring between the lamina lucida and lamina densa of the basement membrane. The severity of these changes increased in a dose‐dependent manner. Morphologically, the IPPSF appeared similar to human skin exposed to HD with the formation of macroscopic blisters and microscopic vesicles. In conclusion, the IPPSF appears to be an appropriate in vitro model with which to study the pathogenesis of vesicant‐induced toxicity. Microsc. Res. Tech., 37:229–241, 1997. © 1997 Wiley‐Liss, Inc.