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Research article1999Peer reviewed

Cytotoxicity of four trichothecenes evaluated by three colorimetric bioassays

Widestrand, Johan; Lundh, Torbjörn; Lindberg, Jan Erik; Pettersson, Hans

Abstract

The application of cell culture technique for screening of low concentrations of Fusarium mycotoxins was examined. Three colorimetric bioassays were used to determine the cytotoxicity of the trichothecenes T-2 toxin (T-2), HT-2 toxin (HT-2), deoxynivalenol (DON) and nivalenol (NIV) to 3T3 mouse fibroblasts (3T3 cells). The bioassays assess DNA synthesis (incorporation of 5-bromo-2'-deoxyuridine; BrdU), metabolic activity (cleavage of 3-(4,5-dimethyltiazol-2-yl)-2,5-diphenyltetrazolium bromide; MTT) and cell membrane damage (release of lactate dehydrogenase; LDH), respectively. The BrdU bioassay was the most sensitive and the IC50 values (50% response compared to untreated cells) of T-2, HT-2, DON and NIV were 4.6, 13, 263 and 365 ng/ml, respectively. At the same toxin concentrations used in the BrdU bioassay, only T-2 and HT-2 were toxic enough to obtain IC50 values using the MTT bioassay. The IC50 values for T-2 and HT-2 were 12 and 68 ng/ml, respectively. When determined by the LDH bioassay, the IC50 values of T-2 and HT-2 were 18 and 42 ng/ml, respectively. At the tested concentrations, DON and NIV had a minor effect on the 3T3 cells when evaluated by the MTT and LDH bioassays. The BrdU bioassay in combination with 3T3 cells was found to be a suitable method for determination of trichothecene-induced toxicity at low concentrations.

Keywords

bioassay; BrdU; cytotoxicity; LDH; MTT; trichothecenes

Published in

Mycopathologia
1999, Volume: 147, number: 3, pages: 149-155
Publisher: KLUWER ACADEMIC PUBL