Electrochemical monitoring of hydrogen peroxide released from leucocytes on horseradish peroxidase redox polymer coated electrode chip

Kumi Y. Inoue, Kosuke Ino, Hitoshi Shiku, Shigenobu Kasai, Tomoyuki Yasukawa, Fumio Mizutani, Tomokazu Matsue

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

We developed an electrochemical-sensing device for continuous monitoring extracellular hydrogen peroxide (H2O2). The device consists of an indium-tin-oxide electrode coated with osmiumpolyvinylpyridine gel polymer containing horseradish peroxidase (Os-HRP) and a poly-dimethyl siloxane well to house the cells on the chip. Granulocyte-like differentiated HL-60 cells were accommodated in the well and stimulated with phorbol 12-myristate 13-acetate (PMA), which triggered the generation of H2O2. The extracellular H2O2 released from the cells was enzymatically reduced at the Os-HRPmodified electrode chip using Os(II) as an electron donor, resulting in reduction current responses by the device. The reduction current increased immediately upon PMA stimulation and this current transient was similar to that obtained by conventional chemiluminescence assays using sodium luminol. Apocynin, an inhibitor of NADPH oxidase activation, eliminated both the electrochemical and chemiluminescence signals. On the other hand, superoxide dismutase (SOD) increased the amperometric signals and catalase (CAT) decreased, whereas SOD decreased luminescence emission and CAT did not. These results were in accordance with the expected reaction mechanism, and strongly indicate that this new electrochemical-sensing device successfully detects extracellular H2O2 production.

Original languageEnglish
Pages (from-to)1723-1728
Number of pages6
JournalBiosensors and Bioelectronics
Volume25
Issue number7
DOIs
Publication statusPublished - 2010 Mar 15

Keywords

  • Amperometory
  • Chemiluminescence
  • Neutrophils
  • Osmium wired peroxidase polymer
  • Reactive oxygen species

ASJC Scopus subject areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

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