Assessment of the Protective Effect of Hexamethylene Tetramin on HF2FF Cell Line Exposed to Sulfur Mustard

Sulfur mustard (HD) as a very dangerous chemical weapon has been used and probably will be used. In spite of widespread research over several decades, no satisfactory prophylactic or treatment regimen has evolved for HD.

In this study, the HF2FF cell line was used as a model of human fibroblastic skin cells to study the toxicity of HD and protective effect of HMT against HD. The viability of cells as an important evaluating parameter was determined by neutral red (NR) dye retention and gentian violet (GV) assays as indicates of cell viability. The ability of HMT to protect HF2FF cells against the effects of an LC5O challenge dose of HD was assessed at different situations.

Application of different concentrations of HD showed that in concentrations excess of 30 jilVi, the onset of HD toxicity is developed rapidly and the viability of cells declined rapidly and dose dependently. HD at concentration of 1000 1iM reduced the viability of cells about 70% and 80% after 5 mm and 1 h, respectively (control 100%). Whereas exposure to 30 1iM of HD even after 1 h declined cell viability only 18%. Dose response curve was used to determine the suitable concentration of HD in cell culture to induce 50% cell death (LC5O) as 180 p]V1 forl5 mm. The concentration of 10 1iIVl of HMT alone did not have any significant effect on cell viability. Pretreatment of cells with HMT before HD exposure could reduce cell death significantly up to 36%. Nonetheless, simultaneous application of HD and HMT was less effective and reduced cell death 23% and 28.5% using GV and NR assays, respectively. HMT post-treatment after HD exposure had no prominent effect to prevent cell death.

The results indicate that HMT can protect these cells against the toxic effect of HD, though it must be present at the time of HD challenge. Application of HMT after HD exposure probably cannot prevent cell death. To evaluate the efficacy of HMT for prevention of HD toxicity, in-vivo studies are warranted. HF2FF cells were found to be a valuable experimental model for studying the toxicity of HD, other skin damaging agents and screening of other compounds with potential therapeutic efficacy before in-vivo studies.