Cartilage degradation is fully reversible in the presence of aggrecanase but not matrix metalloproteinase activity.
BACKGROUND
Physiological and pathophysiological cartilage turnover may coexist in articular cartilage. The distinct enzymatic processes leading to irreversible cartilage damage, compared with those needed for continuous self-repair and regeneration, remain to be identified. We investigated the capacity of repair of chondrocytes by analyzing their ability to initiate an anabolic response subsequent to three different levels of catabolic stimulation.


METHODS
Cartilage degradation was induced by oncostatin M and tumour necrosis factor in articular cartilage explants for 7, 11, or 17 days. The catabolic period was followed by 2 weeks of anabolic stimulation (insulin growth factor-I). Cartilage formation was assessed by collagen type II formation (PIINP). Cartilage degradation was measured by matrix metalloproteinase (MMP) mediated type II collagen degradation (CTX-II), and MMP and aggrecanase mediated aggrecan degradation by detecting the 342FFGVG and 374ARGSV neoepitopes. Proteoglycan turnover, content, and localization were assessed by Alcian blue.


RESULTS
Catabolic stimulation resulted in increased levels of cartilage degradation, with maximal levels of 374ARGSV (20-fold induction), CTX-II (150-fold induction), and 342FFGVG (30-fold induction) (P < 0.01). Highly distinct protease activities were found with aggrecanase-mediated aggrecan degradation at early stages, whereas MMP-mediated aggrecan and collagen degradation occurred during later stages. Anabolic treatment increased proteoglycan content at all time points (maximally, 250%; P < 0.001). By histology, we found a complete replenishment of glycosaminoglycan at early time points and pericellular localization at an intermediate time point. In contrast, only significantly increased collagen type II formation (200%; P < 0.01) was observed at early time points.


CONCLUSIONS
Cartilage degradation was completely reversible in the presence of high levels of aggrecanase-mediated aggrecan degradation. After induction of MMP-mediated aggrecan and collagen type II degradation, the chondrocytes had impaired repair capacity.
Karsdal
Morten A
MA
Nordic Bioscience A/S, Herlev Hovedgade 207, DK-2730 Herlev, Denmark. mk@nordicbioscience.com
Madsen
Suzi H
SH
Christiansen
Claus
C
Henriksen
Kim
K
Fosang
Amanda J
AJ
Sondergaard
Bodil C
BC
D000818:Animals; D002358:Cartilage, Articular / Q000201:enzymology* / Q000473:pathology*; D002417:Cattle; D024043:Collagen Type II / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism*; D004789:Enzyme Activation / Q000502:physiology; D006801:Humans; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D009924:Organ Culture Techniques; D011509:Proteoglycans / Q000378:metabolism
D024043:Collagen Type II; D011509:Proteoglycans; D010450:Endopeptidases; D020782:Matrix Metalloproteinases; C083641:aggrecanase