J Korean Med Sci.  2015 Sep;30(9):1246-1252. 10.3346/jkms.2015.30.9.1246.

Age-related NADPH Oxidase (arNOX) Activity Correlated with Cartilage Degradation and Bony Changes in Age-related Osteoarthritis

• Kim MJ
• Kim HJ
• Hong YH
• Lee CK
• Kim YW
• Shon OJ
• Song IH

• KMID: 2344148
• DOI: http://doi.org/10.3346/jkms.2015.30.9.1246

Abstract

The purpose of this study was to investigate the age-related NADPH oxidase (arNOX) activity in patients with age-related knee osteoarthritis (OA). Serum and cartilage arNOX activities were determined using an oxidized ferricytochrome C reduction assay. Full-thickness knee joint cartilages obtained through total knee replacement surgery were graded according to the Outerbridge (OB) classification. Radiographic severity of OA was determined on Knee X-rays according to the Kellgren-Lawrence (K/L) grading system. Cartilage beta-galactosidase, HIF-1alpha, and GLUT-1 expression levels were evaluated as markers for tissue senescence, hypoxia, and glycolysis. Higher arNOX activities occurred with higher levels of cartilage beta-galactosidase, HIF-1alpha, and GLUT-1 (P = 0.002). arNOX activity in cartilages with surface defects (OB grade II, III) was higher than in those without the defects (OB grade 0, I) (P = 0.012). Cartilage arNOX activity showed a positive correlation with serum arNOX activity (r = -0.577, P = 0.023). Serum arNOX activity was significantly higher in the OA subgroup with bilateral ROA than in the OA with no or unilateral ROA (2.449 +/- 0.81, 2.022 +/- 0.251 nM/mL, respectively, P = 0.019). The results of this study demonstrate that OA itself is not a cause to increase arNOX activities, however, arNOX hyperactivity is related to a high degree of cartilage degradation, and a high grade and extent of ROA in age-related OA.

Keyword

ENOX1 Protein, Human; Age-related NOX; Aging; Osteoarthritis; Cartilage

MeSH Terms

Biomarkers/metabolism
Cartilage Diseases/*enzymology
Cartilage, Articular/*enzymology
Enzyme Activation
Female
Humans
Male
Middle Aged
NADH, NADPH Oxidoreductases
Osteoarthritis, Knee/*diagnosis/*enzymology
Osteoporosis/*diagnosis/*enzymology
Reproducibility of Results
Sensitivity and Specificity
Statistics as Topic
Biomarkers
NADH, NADPH Oxidoreductases

Figure

• 

  Fig. 1 Grades and senescence markers of cartilage from knee OA patients. (A) OA cartilages were classified into OB grade 0 to IV according to the degree of cartilage defect. (B) β-galactosidase staining of OA cartilages showed senescent chondrocytes (bright blue cytoplasmic staining) increasing in cartilages of the higher OB grade. (C) HIF-1α and GLUT-1 expressions showed a significant up-regulation in cartilages of the higher OB grade. The changes of cartilage senescence were consistent with cartilage arNOX hyperactivity in OA cartilages. *P < 0.05; †P < 0.01.

• 

  Fig. 2 arNOX activities of serum in healthy control subjects and those of serum and cartilage in OA patients. (A) Ferricytochrome C reduction assay showed a significantly different activity in serum between a healthy younger man and an older woman. (B) Serum arNOX activity showed a steady increase in correlation with age in healthy control subjects. (C) Serum arNOX activity in OA patients increased in correlation with the K/L grade of both knees. (D) Serum arNOX activity was significantly higher in patients with bilateral ROA as compared to those with no or unilateral ROA. (E) Cartilage arNOX activity showed a strong positive correlation with serum arNOX activity in OA patients. (F) Cartilage arNOX activity was more active in cartilages with surface defects (OB grade II, III) than in those without defects (OB grade 0, I). *P < 0.05; †P < 0.01.

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