Document Detail

Simultaneous use of electrochemistry and chemiluminescence to detect reactive oxygen species produced by human neutrophils.
MedLine Citation:
PMID:  18782628     Owner:  NLM     Status:  MEDLINE    
A novel approach for the simultaneous optical and electrochemical detection of biologically produced reactive oxygen species has been developed and applied. The set-up consists of a luminol-dependent chemiluminescence assay combined with two amperometric biosensors sensitive to superoxide anion radicals (O(2)(-)) and hydrogen peroxide (H(2)O(2)), respectively. The method permits direct, real-time in vitro determination of both extra- and intracellular O(2)(-) and H(2)O(2) produced by human neutrophil granulocytes. The rate of O(2)(-) production by stimulated neutrophils was calculated to about 10(-17)mol s(-1) per single cell. With inhibited NADPH oxidase, a distinct extracellular release of H(2)O(2) instead of O(2)(-) was obtained from stimulated neutrophils with the rate of about 3 x 10(-18)mol s(-1) per single cell. When the H(2)O(2) release was discontinued, fast H(2)O(2) utilisation was observed. Direct interaction with and possibly attachment of neutrophils to redox protein-modified gold electrodes, resulted in a spontaneous respiratory burst in the population of cells closely associated to the electrode surface. Hence, further stimulation of human neutrophils with a potent receptor agonist (fMLF) did not significantly increase the O(2)(-) sensitive amperometric response. By contrast, the H(2)O(2) sensitive biosensor, based on an HRP-modified graphite electrode, was able to reflect the bulk concentration of H(2)O(2), produced by stimulated neutrophils and would be very useful in modestly equipped biomedical research laboratories. In summary, the system would also be appropriate for assessment of several other metabolites in different cell types, and tissues of varying complexity, with only minor electrode modifications.
Sergey Shleev; Jonas Wetterö; Karl-Eric Magnusson; Tautgirdas Ruzgas
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-08-20
Journal Detail:
Title:  Cell biology international     Volume:  32     ISSN:  1095-8355     ISO Abbreviation:  Cell Biol. Int.     Publication Date:  2008 Dec 
Date Detail:
Created Date:  2008-11-18     Completed Date:  2009-01-09     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9307129     Medline TA:  Cell Biol Int     Country:  England    
Other Details:
Languages:  eng     Pagination:  1486-96     Citation Subset:  IM    
Biomedical Laboratory Science, Health and Society, Malmö University, 20506 Malmö, Sweden.
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MeSH Terms
Biosensing Techniques / instrumentation,  methods*
Cells, Cultured
Chemiluminescent Measurements / instrumentation,  methods*
Electrochemical Techniques / instrumentation,  methods*
Electrodes / standards
Hydrogen Peroxide / analysis,  metabolism
Immunity, Innate / physiology
Neutrophils / chemistry*,  metabolism*
Optics and Photonics / instrumentation,  methods
Oxygen / metabolism
Reactive Oxygen Species / analysis,  metabolism*
Superoxides / analysis,  metabolism
Reg. No./Substance:
0/Reactive Oxygen Species; 11062-77-4/Superoxides; 7722-84-1/Hydrogen Peroxide; 7782-44-7/Oxygen

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