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EVALUATION OF A TISSUE ENGINEERED HUMAN SKIN EPIDERM 3-D CULTURE AS A MODEL TO STUDY IRRITATION AFTER JET FUEL EXPOSURE.

Chatterjee, A., Babu, R., Singh, M. Florida A&M University, Tallahassee, FL, USA.
Abstract

This study by researchers at Florida A&M University demonstrated that MatTek’s EpiDerm in vitro human skin tissue equivalent is a “valuable model to evaluate the skin irritation potential of jet fuels and hydrocarbon chemicals”. The purpose of the present study was to evaluate human skin Epiderm™ 3-D culture, EPI-200 (MatTek Corp, Ashland, MA) as a model to study the skin irritation of various jet fuels. Upon receipt, EPI-200 tissues were placed in six-well plates containing maintenance medium (EPI-100-NMM) and incubated for 24-72h. The cultures were subjected to different doses (5, 10, 50 µl) of JP-8 and JP-8+100. After each exposure, the tissue viability was measured by MTT assay and the morphology was studied by hematoxylin and eosin (H&E) staining of histological sections. The expression of IL-1a in the tissue culture media was determined by enzyme immunoassay (EIA). The EPI-200 exposures showed an interesting dose dependent relationship between IL-1a release and cell viability. At the lowest dose (5 µl) of JP-8 and JP-8+100, there was no cell kill at 24 h, but at 48 and 72 h both the chemicals reduced the tissue viability to 61-65 %, with no significant difference between the two time points. JP-8 and JP-8+100 exposures at 24h significantly increased the expression of IL-1a to 13 and 51 folds respectively over the control. This finding was very interesting considering the fact that there was no cell kill detected after 24 h. At 48 and 72h the IL-1a level was lower than at 24h which was due to reduction in the cell viability. After 24h, 10 µl of JP-8 and JP-8+100 exposures demonstrated around 45-50% cell viability while 50 µl of the jet fuels reduced the cell viability to zero. Control tissues were 100% viable at all time points. H&E staining of the tissues after 24h indicated that the jet fuels disrupted the barrier function of stratum corneum. The effect was more severe after 48h, while after 72h the tissue started losing their integrity. The findings with EPI200 correlate well with our earlier findings from skin irritation studies conducted on CD Hairless rats with JP8 and JP8100. Overall, our experiments demonstrate that EPI-200 can be used as a valuable model to evaluate the skin irritation potential of jet fuels and hydrocarbon chemicals.

Keywords

3-D, Contact dermatitis, Dermal corrosion, Dermal phototoxicity studies, Dodecane, EPI-200, EpiDerm, Hazardous materials, Histo-pathological, Histological studies, IL-1a, Immune suppression, Irritation, Jet fuel exposure, MTT, MTT assay, NHEK, Nonane, Skin irritation, TNF-a, Tetradecane, Toxicity, Toxicological

Materials Tested

Dodecane, JP-8, JP-8+100, Jet fuel, Jet-A, Nonane, Tetradecane

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