Profile of glycosaminoglycan‐degrading glycosidases and glycoside sulfatases secreted by human articular chondrocytes in homeostasis and inflammation

• Published in: Arthritis and rheumatism 2000-06, Vol.43 (6), p.1307-1314
• Main Authors: Shikhman, Alexander R., Brinson, Diana C., Lotz, Martin
• Format: Article
• Language: English
• Subjects: Apoptosis - physiology; beta-N-Acetylhexosaminidases - metabolism; Biological and medical sciences; Cartilage, Articular - drug effects; Cartilage, Articular - enzymology; Cartilage, Articular - pathology; Cells, Cultured; Chondrocytes - drug effects; Chondrocytes - enzymology; Chondrocytes - physiology; Diseases of the osteoarticular system; Extracellular Space - enzymology; Glycosaminoglycans - metabolism; Glycoside Hydrolases - metabolism; Glycosides - metabolism; Homeostasis - physiology; Humans; Hydrogen-Ion Concentration; Inflammation Mediators - pharmacology; Interleukin-1 - pharmacology; Medical sciences; Osteoarthritis; Osteoarthritis - enzymology; Osteoarthritis - pathology; Reference Values; Space life sciences; Sulfatases - metabolism; Synovial Fluid - enzymology
• ISSN: 0004-3591; 1529-0131
• DOI: 10.1002/1529-0131(200006)43:6<1307::AID-ANR13>3.0.CO;2-3

Objective To determine enzymatic activities of the 8 key glycosaminoglycan‐degrading glycosidases and glycoside sulfatases in cultured human articular chondrocytes and in synovial fluid from patients with osteoarthritis. Methods The following enzymes were analyzed: hexosaminidase and its isoenzyme A, N‐acetyl‐α‐D‐glucosaminidase, β‐galactosidase, β‐glucuronidase, α‐L‐iduronidase, aryl sulfatase, and galactose‐6‐sulfate sulfatase. Activity of the selected enzymes was analyzed by fluorometry with the aid of 4‐methylumbelliferryl derivatives of the appropriate monosaccharides. Results Hexosaminidase was found to be the dominant enzyme released by chondrocytes into the extracellular compartment. Stimulation of chondrocytes with interleukin‐1β resulted in a selective increase of the extracellular hexosaminidase activity and, to a lesser degree, of the extracellular β‐galactosidase activity, without significant changes in the activity of the other studied enzymes. Analysis of the pH dependency of the enzymatic activities revealed that even at neutral pH, hexosaminidase expressed a measurable activity, much higher than the activity of the other studied enzymes. Chondrocyte apoptosis did not result in increased extracellular glycosidase activities, including hexosaminidase activity. The spectrum of glycosidase and glycoside sulfatase activities in the synovial fluid from patients with osteoarthritis was similar to that in cultured human articular chondrocytes. Conclusion These data support the concept that lysosomal glycosidases, in particular hexosaminidase, represent a distinct subset of cartilage matrix–degrading enzymes that are activated by proinflammatory stimuli.