Hydroxyl radical formation in chondrocytes and cartilage as detected by electron paramagnetic resonance spectroscopy using spin trapping reagents

Chondrocytes have been shown to produce superoxide and hydrogen peroxide, suggesting possible formation of hydroxyl radical in these cells. In this study, we used electron spin resonance/spin trapping technique to detect hydroxyl radicals in chondrocytes. We found that hydroxyl radicals could be det...

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Bibliographic Details
Published in:Free radical research 1998-01, Vol.29 (3), p.177-187
Main Authors: Tiku, Moti L., Yan, Yong Ping, Chen, Kuang Yu
Format: Article
Language:English
Subjects:
Animals
arthritis
cartilage
Cartilage - chemistry
Cartilage - drug effects
Cartilage - metabolism
Chondrocytes
Chondrocytes - chemistry
Chondrocytes - drug effects
Chondrocytes - metabolism
electron spin resonance
Electron Spin Resonance Spectroscopy
Ethanol
Female
Humans
Hydroxyl Radical - analysis
Hydroxyl Radical - metabolism
hydroxyl radicals
Interleukin-1 - pharmacology
Male
Nitrogen Oxides - analysis
Nitrogen Oxides - chemistry
Osteoarthritis - metabolism
Osteoarthritis - pathology
oxygen intermediates
Pyridines
Rabbits
Spin Labels
Spin Trapping
Synovial Fluid
Tetradecanoylphorbol Acetate - pharmacology
Tumor Necrosis Factor-alpha - pharmacology
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Summary:Chondrocytes have been shown to produce superoxide and hydrogen peroxide, suggesting possible formation of hydroxyl radical in these cells. In this study, we used electron spin resonance/spin trapping technique to detect hydroxyl radicals in chondrocytes. We found that hydroxyl radicals could be detected as α-hydroxyethyl spin trapped adduct of 4-pyridyl 1-oxide N-tert-butylnitrone (4-POBN) in chondrocytes stimulated with phorbol 12-myristate 13-acetate in the presence of ferrous ion. The formation of hydroxyl radical appears to be mediated by the transition metal-catalyzed Haber-Weiss reaction since no hydroxyl radical was detected in the absence of exogenous iron. The hydroxyl radical formation was inhibited by catalase but not by superoxide dismutase, suggesting that the hydrogen peroxide is the precursor. Cytokines, IL-1 and TNF enhanced the hydroxyl radical formation in phorbol 12-myristate 13-acetate treated chondrocytes. Interestingly, hydroxyl radical could be detected in unstimulated fresh human and rabbit cartilage tissue pieces in the presence of iron. These results suggest that the formation of hydroxyl radical in cartilage could play a role in cartilage matrix degradation.
ISSN:1071-5762
1029-2470
DOI:10.1080/10715769800300211