Abstract
ABSTRACTThis paper reports the development and application of a biosensor for the amperometric determination of galactose in the presence of human hepatocellular carcinoma cells with and without a hepatotoxic agent. The biosensor was fabricated by drop-coating 1.5% cellulose acetate on a 3 × 3 mm screen-printed carbon electrode followed by depositing 2 U of galactose oxidase. The electrodes dimensions were reduced to 3 × 0.5 mm before measurements. Hepatocellular carcinoma cells were utilized for in vitro toxicity testing by evaluating the effect of paracetamol on galactose uptake. The amperometric responses to galactose indicated that the inhibition of uptake was directly proportional to the concentration of paracetamol following 24 h of exposure to the hepatocellular carcinoma cells. These results demonstrate that the fabricated biosensor may be used for the real-time monitoring of cell metabolism and toxicity.
Highlights
There is a considerable interest in using cell-based assays, ie. experiments based on the use of live cells, for screening the toxicity of chemical compounds, in the pharmaceutical industry
Hepatotoxicity is a major concern in drug testing as most toxic effects observed are by drugs that are metabolised by the liver; liver cells have been utilised by many drug discovery and development laboratories (Anderson et al 1996; Li et al 1999; Ni et al 2001; Valentin et al 2001)
The galactose biosensor exhibited a sensitivity of 7.2 μAmM-1cm-2, a linear range up to 9.52 mM and a CV of 1.2 %
Summary
There is a considerable interest in using cell-based assays, ie. experiments based on the use of live cells, for screening the toxicity of chemical compounds, in the pharmaceutical industry. Experiments based on the use of live cells, for screening the toxicity of chemical compounds, in the pharmaceutical industry These include a variety of techniques for measuring cell proliferation, morphology, viability, cytotoxicity, and motility (Keese and Giaever 1994; Riss, O'Brien and Morvec 2003; Nabhan 2003). Whereas cell viability assays generally involve measuring the number of live cells, cytotoxicity assays tend to evaluate the number of dead cells Both types of cell-based assay are routinely used for drug discovery using high-throughput screening, environmental assessment of relevant chemical compounds, and as biosensors for cellular behaviour analysis (Ehret et al 1997; White 2000; Shuileabhain et al 2004). Hepatotoxicity is a major concern in drug testing as most toxic effects observed are by drugs that are metabolised by the liver; liver cells have been utilised by many drug discovery and development laboratories (Anderson et al 1996; Li et al 1999; Ni et al 2001; Valentin et al 2001)
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