Effects of Subtoxic Doses of Aflatoxin B ₁ : An In Vitro Assessment of Micronucleus Assay and Induction of Neutrophil Extracellular Traps

Aflatoxin B₁ (AFB₁) is a mycotoxin mainly produced by Aspergillus flavus and Aspergillus parasiticus. The genotoxic capacity of AFB₁ has been demonstrated in a range of genotoxicity assays. The aim of the present study was to evaluate the cytotoxic and genotoxic effects of AFB₁ and fumonisin B₁ (FB₁) mixtures on hepatocellular carcinoma cell line (HepG2), as well as the inflammatory effects on human neutrophils of AFB₁ at environmentally relevant concentrations. The cytotoxicity was evaluated using the MTT assay. The cytostatic and genotoxic effects were evaluated using the micronucleus technique, and the inflammatory effect of AFB₁ was determined by fluorescence microscopy to evaluate neutrophil extracellular traps (NETs). Additionally, we have explored the response of the system in a coexposure treatment with FB₁. Significant increases in micronucleus frequency, apoptosis, and necrosis were observed after treatment with 10-μM AFB₁ or 10-μM AFB₁ + 50-μM FB₁. FB₁ did not have a synergistic effect with AFB₁. A relative impairment of the genetic material reflected by nucleoplasmic bridges, nuclear budding, and donut-shaped nuclei was observed. On the other hand, the assessment of the inflammatory response showed that AFB₁ induces the formation of extracellular traps, apparently dose dependent.