3D "honeycomb" cell/carbon nanofiber/gelatin methacryloyl (GelMA) modified screen-printed electrode for electrochemical assessment of the combined toxicity of deoxynivalenol family mycotoxins.

A "honeycomb" electrochemical biosensor based on 3D printing was developed to noninvasively monitor the viability of 3D cells and evaluate the individual or combined toxicity of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), and 15-acetyldeoxynivalenol (15-ADON). Carbon nanofiber (CN)/gelatin methacryloyl (GelMA) conductive composite hydrogel with strong processability was printed on 8-channel screen-printed carbon electrodes (SPCEs) to maintain cell viability and form tight cell-to-cell contacts. A "3D honeycomb" printing infill pattern was selected in the construction of the biosensors to improve conductivity. Based on 3D printing technology, the electrochemical biosensor can prevent manual error and provide for high-throughput detection. Electrochemical impedance spectroscopy (EIS) was used to evaluate mycotoxin toxicity. The EIS response decreased with the concentration of DON, 3-ADON and 15-ADON in the range of 0.1-10, 0.05-100, and 0.1-10 μg/mL, respectively, with a limit of detection of 0.07, 0.10 and 0.06 μg/mL, respectively. Mycotoxin interactions were analyzed using the isobologram-combination index (CI) method. The electrochemical cytotoxicity evaluation result was confirmed by biological assays. Therefore, a novel method for evaluating the combined toxicity of mycotoxins is proposed, which exhibits potential for application to food safety and evaluation.