Evaluation of cytogenetic and DNA damage induced by the antibacterial drug, trimethoprim

Trimethoprim, a widely used antimicrobial drug was tested for its effect on the level of nuclear DNA damage in cultured peripheral blood lymphocytes in terms of chromosome and DNA alterations. The extent of cytogenetic damage, expressed as chromosome breakage and chromosome loss, was evaluated employing the cytokinesis block micronucleus method (CBMN) in cultured peripheral blood lymphocytes coupled with fluorescence in situ hybridization (FISH) using a digoxigenin-labelled alphoid DNA probe specific for the centromere of all human chromosomes. The DNA breakage level was evaluated by the Comet assay. Cultures were set up by using blood samples from two healthy donors. A range of concentrations of the test agent (from 1 to 100 μg/ml) was used for the micronuclei (MN) frequency and cytogenetic origin of MN. For the Comet assay the range of doses used was from 0.5 to 150 μg/ml. From the results obtained it appears that this antifolic agent has a significant clastogenic potential, as detected by a dose-dependent increase of the incidence of C-MN and significantly greater than control levels at the highest concentrations tested (25,100 μg/ml). In addition, the results obtained in the Comet assay also show that trimethoprim induces a dose-dependent increase in the level of DNA breakage, this increase attaining statistical significance at the highest concentrations tested (25, 100, 150 μg/ml), which would confirm its genotoxicity.