Application of screen-printed microband biosensors incorporated with cells to monitor metabolic effects of potential environmental toxins

Abstract

Microband biosensors were fabricated from a screen-printed water-based carbon ink containing cobalt phthalocyanine redox mediator and glucose oxidase or lactate oxidase enzyme. The microbiosensors were characterised for their ability to monitor ferrocyanide and H2O2 in phosphate buffer solution: sigmoidal cyclic voltammograms, high current density values and steady-state amperometric responses confirmed the existence of radial-diffusion limiting microelectrode behaviour. The lactate microband biosensors were then used, in conjunction with a screen-printed Ag/AgCl reference and platinum counter electrode, tomonitor lactate levels in culture medium, with a linear rangeof 0.5–5 mM, sensitivity of 20 nA.mM−1, and dynamic range up to >9 mM. The lactate microband biosensors could operate continuously in culture medium over extended times(up to 24 h) at 37 °C. These biosensors were then applied todetect changes in lactate release from cultured cells in response to toxic challenge: m-dinitrobenzene (500 μM)caused a reduction in lactate production by high-passage number HepG2 single cells; D-galactosamine (20 mM) induced release of lactate by HepG2 spheroid cultures. This novel use of microband biosensors in cell culture has the potential for further application in toxicity monitoring, in both environmental and pharmaceutical areas.