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RESEARCH PRODUCT

Toxicity evaluation of individual and mixed enniatins using an in vitro method with CHO-K1 cells

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Enniatins (ENs) A, A1, B and B1 are produced by Fusarium species. They are known as emerging fusario- toxins, and can cause outbreaks in both humans and animals. ENs elicits a wide range of different biolog- ical properties and toxicological effects, and their co-occurrence may enhance the extent of these hazards. As the potential toxins reach in vitro cells in the same way as they would in vivo, cytotoxicity was studied with CHO-K1, which is considered one of the most sensitive cell lines for preliminary screen- ing of cytotoxicity studies. In this study, individual cytotoxic effects of ENs were evaluated by MTT assay after exposing ENs to CHO-K1 cells for 24, 48, and 72 h. The IC50 values obtained for ENs A, A1, B and B1 ranged from >7.5 to 2.83 ± 0.49, from 8.8 ± 2.29 to 1.65 ± 0.06, from 11.0 ± 2.65 to 2.80 ± 0.16 and from 4.53 ± 1.23 to 2.47 ± 0.29 lM, respectively. The ENs cytotoxic interaction was evaluated by the isobolo- gram method. The IC50 values for ENs combination ranged from 0.44 ± 0.15 (ENs A1 +B 1) to 0.97 ± 0.48 (ENs A1 +B+B 1). The binary combinations ENs A + B1, ENs A1 + B and ENs B + B1 showed additive effect thought all concentrations tested. Sinergistic effect of combined ENs A + A1 ,A + B, A 1 +B 1 ,A + A 1 +B , A+A 1 +B 1 , A +B+B 1 and A1 +B+B 1 at higher concentrations occurred. Synergism effect (CI from 0.37 ± 0.08 to 0.74 ± 0.22) was observed at higher concentrations with binary and tertiary combinations of EN A, while antagonism effect was obtained at lower concentrations for ENs A + A1 +B 1 (CI = 2.67 ± 1.32) and ENs A1 +B+B 1 (CI = 3.14 ± 1.91). These results provide quantitative evidence that ENs cytotoxicity depend on their concentrations, and also on their combination with other mycotoxins. 2012 Elsevier Ltd. All rights reserved.