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Presence of methenamine/glutathione mixtures reduces the cytotoxic effect of sulphur mustard on cultured SVK-14 human keratinocytes in vitro

Chemical and Biological Defence Porton Down, Salisbury, Wiltshire, SP4 0JQ, UK

CD Lindsay

Chemical and Biological Defence Porton Down, Salisbury, Wiltshire, SP4 0JQ, UK

DG Upshall

Chemical and Biological Defence Porton Down, Salisbury, Wiltshire, SP4 0JQ, UK

1 The basal epidermal keratinocytes of the skin are a main target for the vesicating agent, sulphur mustard (SM). A human keratinocyte cell line (SVK-14) has been used to model the effects of SM on the basal epidermal keratinocytes and subsequently to test the efficacy of potential prophylactic compounds in reducing the SM-induced cytotoxicity.

2 The cultures were pretreated with mixtures of methe namine (HMT) and glutathione (GSH) for 1 h prior to exposure to 10 µM SM. The viability of the cultures was then assessed using neutral red (NR) dye uptake and crystal violet DNA staining assays at 24 h intervals post exposure.

3 Pretreatment led to a 1.9 fold increase in culture viability (NR assay) compared to those exposed to SM only, and a 2.3 fold increase in cell number (crystal violet assay). Photomicrography showed that pre treatment preserved the morphology of the cultured cells and maintained their mitotic activity whereas those exposed to SM only show non-proliterative cultures with extensive cellular damage.

4 The results of this study show that it is possible to protect mitotically active cultures from the effects of SM, however the measures must be in place prior to SM exposure.

Key Words: human keratinocytes • sulphur mustard • glutathione • methenamine

Human & Experimental Toxicology, Vol. 16, No. 5, 247-253 (1997)
DOI: 10.1177/096032719701600502


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