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Title: Effect of sulfur mustard on mast cells in hairless guinea pig skin

Abstract

The skin of 24 anesthetized hairless guinea pigs was exposed to saturated sulfur mustard (bis-2-chloroethyl sulfide; HD) for 5 and 7 minutes using 14-mm diameter vapor cups. Animals were euthanatized 24 hours after exposure and skin specimens taken for morphometric evaluation of granulated mast cells with an image analysis system (IAS). Tissue specimens were processed in paraffin, sectioned at 5 microns and stained with Unna's stain for mast cells. The number of granulated mast cells and the area occupied by mast cell granules was determined. There were significantly fewer mast cells (p < 0.05) in either HD exposure group than in sham-exposed animals, with significantly fewer mast cells in the 7-minute than the 5-minute HD group. There were also significantly smaller areas occupied by granules in either HD exposure group than in sham-exposed animals. HD-induced lesions in the hairless guinea pig have shown signs of an inflammatory response, and with their granules of vasoactive histamine, mast cells might be expected to play a role in HD-induced injury. Changes in mast cells exposed to low sulfur mustard levels, as detected by an IAS, may serve as an early marker for cutaneous damage, which might not be as easily determined with routinemore » light microscopy.« less

Authors:
; ;
Publication Date:
Research Org.:
Army Medical Research Inst. of Chemical Defense, Aberdeen Proving Ground, MD (United States)
OSTI Identifier:
7239946
Alternate Identifier(s):
OSTI ID: 7239946
Report Number(s):
AD-P-008756/9/XAB
Resource Type:
Technical Report
Resource Relation:
Other Information: This article is from 'Proceedings of the Medical Defense Bioscience Review (1993) Held in Baltimore, Maryland on 10-13 May 1993. Volume 1, AD-A275 667, p41-48
Country of Publication:
United States
Language:
English
Subject:
45 MILITARY TECHNOLOGY, WEAPONRY, AND NATIONAL DEFENSE; 63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; CHEMICAL WARFARE AGENTS; TOXICITY; WEAPONS 450600* -- Military Technology, Weaponry, & National Defense-- Chemical & Biological-- (1990); 560300 -- Chemicals Metabolism & Toxicology

Citation Formats

Graham, J.S., Bryant, M.A., and Braue, E.H.. Effect of sulfur mustard on mast cells in hairless guinea pig skin. United States: N. p., 1993. Web.
Graham, J.S., Bryant, M.A., & Braue, E.H.. Effect of sulfur mustard on mast cells in hairless guinea pig skin. United States.
Graham, J.S., Bryant, M.A., and Braue, E.H.. Thu . "Effect of sulfur mustard on mast cells in hairless guinea pig skin". United States. doi:.
@article{osti_7239946,
title = {Effect of sulfur mustard on mast cells in hairless guinea pig skin},
author = {Graham, J.S. and Bryant, M.A. and Braue, E.H.},
abstractNote = {The skin of 24 anesthetized hairless guinea pigs was exposed to saturated sulfur mustard (bis-2-chloroethyl sulfide; HD) for 5 and 7 minutes using 14-mm diameter vapor cups. Animals were euthanatized 24 hours after exposure and skin specimens taken for morphometric evaluation of granulated mast cells with an image analysis system (IAS). Tissue specimens were processed in paraffin, sectioned at 5 microns and stained with Unna's stain for mast cells. The number of granulated mast cells and the area occupied by mast cell granules was determined. There were significantly fewer mast cells (p < 0.05) in either HD exposure group than in sham-exposed animals, with significantly fewer mast cells in the 7-minute than the 5-minute HD group. There were also significantly smaller areas occupied by granules in either HD exposure group than in sham-exposed animals. HD-induced lesions in the hairless guinea pig have shown signs of an inflammatory response, and with their granules of vasoactive histamine, mast cells might be expected to play a role in HD-induced injury. Changes in mast cells exposed to low sulfur mustard levels, as detected by an IAS, may serve as an early marker for cutaneous damage, which might not be as easily determined with routine light microscopy.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Thu May 13 00:00:00 EDT 1993},
month = {Thu May 13 00:00:00 EDT 1993}
}

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  • We have reported (Proceedings of the 1991 Medical Defense Bioscience Review) the use of a reflectance color meter for the quantification of erythema and a high frequency ultrasound system for the evaluation of edema by measuring skin thickness. This report compares these two techniques with several new methods including laser Doppler flowmetry (LDF) to measure microcirculation, skin elasticity measurements made both parallel (Dermal Torque Meter, DTM) and perpendicular (Cutometer) to the skin surface, and capacitance measurements for evaluating the skin hydration state. The skin of anesthetized hairless guinea pigs was exposed to saturated HD vapor (1.4mg/mi) at 4 sites formore » 3, 5, 7, or 9 minutes. Lesions were evaluated by each of the analytical methods at various times up to 24 hours post-exposure. Results have demonstrated a dose-response relationship between HD vapor exposure and erythema (reflectance color meter), edema (high frequency ultrasound), cutaneous microcirculation (LDF), and skin elasticity measured perpendicular to the skin surface (Cutometer). HD vapor cutaneous exposure had only a marginal effect on skin elasticity parallel (DTM) to the skin and on moisture content. These preliminary results suggest that reflectance color meter, high frequency ultrasound, LDF, and skin elasticity measurements may be useful for determining the efficacy of candidate antivesicant therapies.« less
  • Hypochlorite solutions are thought to be efficacious when used to topically decontaminate intact skin. However, few studies have examined the efficacy of decontamination of chemically contaminated wounds. Therefore, we compared the decontamination efficacy of sodium hypochlorite (0.5% and 2.5% solutions), calcium hypochlorite (0.5% and 2.5% solutions) and sterile water to untreated controls in wounds exposed to sulfur mustard (HD). Anesthetized euthymic hairless guinea pigs (EHGP) (n=6) were exposed to 0.4 LD50 HD in a full-thickness 8 mm surgical biopsy skin defect (i.e., wound). Each animal was subsequently decontaminated, after a two-minute intra-wound exposure to liquid HD, with one of themore » decontamination solutions. Decontamination efficacy was determined by the visual grading of the HD-traumatized wound lesion and by comparison of the expected HD-induced leukocyte suppression. Leukocyte suppression was inconsistent in all animals; therefore, the visual grading was the only viable evaluation method. No significant differences were observed among wounds decontaminated with any of the solutions. However, the skin surrounding undecontaminated (but exposed) control animals showed the least visual pathology. The lesions induced following decontamination are presumed to be due to the mechanical flushing HD onto the peri-lesional skin, or by chemical damage induced by the solution, or HD-solution interaction. Further studies are required to best delineate the optimal decontamination process for HD contaminated wounds.« less
  • Ten anesthetized hairless guinea pigs Crl:IAF(HA)BR were exposed to 10 pi of neat sulfur mustard (HD) in a vapor cup on their skin for 7 min. At 24 h postexposure, the guinea pigs were euthanatized and skin sections taken for histologic evaluation. The skin was fixed using either 10% neutral buffered formalin (NBF), McDowell Trump fixative (4CF-IG), Zenker`s formol-saline (Helly`s fluid), or Zenker`s fluid. Fixed skin sections were cut in half: one half was embedded in paraffin and the other half in plastic (glycol methacrylate). Paraffin-embedded tissue was stained with hematoxylin and eosin; plastic-embedded tissue was stained with Lee`s methylenemore » blue basic fuchsin. Skin was also frozen unfixed, sectioned by cryostat, and stained with pinacyanole. HD-exposed skin was evaluated histologically for the presence of epidermal and follicular necrosis, microblister formation, epidermitis, and intracellular edema to determine the optimal fixation and embedding method for lesion preservation. The percentage of histologic sections with lesions varied little between fixatives and was similar for both paraffin and plastic embedding material. Plastic-embedded sections were thinner, allowing better histologic evaluation, but were more difficult to stain. Plastic embedding material did not infiltrate tissue fixed in Zenker`s fluid or Zenker`s formol-saline. Frozen tissue sections were prepared in the least processing time and lesion preservation was comparable to fixed tissue. It was concluded that standard histologic processing using formalin fixation and paraffin embedding is adequate for routine histopathological evaluation of HD skin lesions in the hairless guinea pig.... Sulfur mustard, Vesicating agents, Pathology, Hairless guinea pig model, Fixation.« less
  • Hypochlorite solutions are thought to be efficacious when used to topically decontaminate intact skin. However, few studies have examined the efficacy of decontamination of chemically contaminated wounds. Therefore, we compared the decontamination efficacy of sodium hypochlorite (0.5% and 2.5% solutions), calcium hypochlorite (0.5% and 2.5% solutions) and sterile water to untreated controls in wounds exposed to sulfur mustard (HD). Anesthetized euthymic hairless guinea pigs (EHGP) (n=6) were exposed to 20 mg/kg (approximately 0.4 LD%) HD in a full-thickness 8 mm surgical biopsy skin defect (i.e., wound). Each animal was subsequently decontaminated, after a two-minute intra-wound exposure to liquid HD, withmore » nothing or one of the decontamination solutions. Decontamination efficacy was determined by the visual grading of the HD-traumatized wound lesion and by comparison of the expected HD-induced leukocyte suppression. Leukocyte suppression was inconsistent in all animals; therefore, the visual grading was the only viable evaluation method. No significant differences were observed among wounds decontaminated with any of the solutions. However, the skin surrounding non-decontaminated (but exposed) control animals showed the least visual pathology. The lesions induced following decontamination are presumed to be due to the mechanical flushing of HD onto the peri-lesional skin, or by chemical damage induced by the solution, or ND-solution interaction. Further studies are required to best delineate the optimal decontamination process for HD contaminated wounds.« less
  • Increased proteolytic activity at the dermal-epidermal junction is postulated as being involved in sulfur-mustard-induced cutaneous injury. Homogenates of skin punch biopsy specimens from the skin of hairless guinea pig at 6, 9, 12, and 24 h after a 7 min vapor cup exposure to sulfur mustard (HD) demonstrated enhanced proteolytic activity. Homogenates from the biopsy specimens of exposed animals produced from 3 to 10 times the hydrolysis of the chromogenic peptide substrate Chromzym TH (tosyl-gly-pro-arg-p-nitrani-lide) and human elastase substrate N-methoxysuccinyl-ala- ala-pro-val-p-nitranilide than did the homogenates from control samples. In this study HD-increased proteolysis of the TH substrate by extracts ofmore » hairless guinea pig skin biopsies was nearly eliminated by systemic treatment with hydroxyurea and greatly reduced by topical application of the anti-inflammatory compound phenidone. Compounds that reduce HD-increased proteolytic activity, such as phenidone and hydroxyurea, can serve as probes to examine the role of proteolysis in HD-induced pathology. HD-increased proteolysis provides a biochemical correlate for investigating cutaneous exposure to HD. Increased proteolysis may therefore serve as a biomarker for HD-induced pathology that might be used as an index of efficacy for potential treatment compounds.« less