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Application of cyclic biamperometry to viability and cytotoxicity assessment in human corneal epithelial cells

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Abstract

The application of cyclic biamperometry to viability and cytotoxicity assessments of human corneal epithelial cells has been investigated. Electrochemical measurements have been compared in PBS containing 5.0 mM glucose and minimal essential growth medium. Three different lipophilic mediators including dichlorophenol indophenol, 2-methyl-1,4-naphthoquinone (also called menadione or vitamin K3) and N,N,N′,N′-tetramethyl-p-phenylenediamine have been evaluated for shuttling electrons across the cell membrane to the external medium. Transfer of these electrons to ferricyanide in the extra cellular medium results in the accumulation of ferrocyanide. The amount of ferrocyanide is then determined using cyclic biamperometry and is related to the extent of cell metabolic activity and therefore cell viability. To illustrate cytotoxicity assessment of chemicals, hydrogen peroxide, benzalkonium chloride and sodium dodecyl sulfate have been chosen as sample toxins, the cytotoxicities of which have been evaluated and compared to values reported in the literature. Similar values have been reported using colorimetric assays; however, the simplicity of this electrochemical assay can, in principle, open the way to miniaturization onto lab-on-chip devices and its incorporation into tiered-testing approaches for cytotoxicity assessment.

After mediated reduction of ferricyanide by cell components, its reduced form, ferrocyanide, is quantitated using cyclic biamperometry in presence of excess ferricyanide. Concentration of ferrocyanide is then related to the extent of cell metabolic activity for viability/cytotoxicity assessments

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Acknowledgments

Financial support from Ontario Ministry of Education and the Natural Sciences and Engineering Research Council of Canada is gratefully acknowledged. Helpful comments from reviewers of this work are also appreciated.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Waterloo, 200 University Ave W., Waterloo, ON, N2L 3G1, Canada

    Mehdi Rahimi & Susan R. Mikkelsen

  2. School of Optometry, University of Waterloo, Waterloo, ON, N2L 3G1, Canada

    Hyun-Yi Youn & Jacob G. Sivak

  3. Centre for Contact Lens Research/School of Optometry, University of Waterloo, Waterloo, ON, N2L 3G1, Canada

    David J. McCanna

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  1. Mehdi Rahimi
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Correspondence to Mehdi Rahimi.

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Rahimi, M., Youn, HY., McCanna, D.J. et al. Application of cyclic biamperometry to viability and cytotoxicity assessment in human corneal epithelial cells. Anal Bioanal Chem 405, 4975–4979 (2013). https://doi.org/10.1007/s00216-013-6843-z

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  • DOI: https://doi.org/10.1007/s00216-013-6843-z

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