Cartilage degradation is fully reversible in the presence of aggrecanase but not matrix metalloproteinase activity

Morten A Karsdal¹ , Suzi H Madsen¹ , Claus Christiansen¹ , Kim Henriksen¹ , Amanda J Fosang² and Bodil C Sondergaard¹

¹Nordic Bioscience A/S, Herlev Hovedgade 207, DK-2730 Herlev, Denmark

²University of Melbourne Department of Paediatrics and Murdoch Childrens Research Institute, Royal Children's Hospital, Flemington Road, Parkville, 3052, Melbourne, Victoria, Australia

author email corresponding author email

Arthritis Research & Therapy 2008, 10:R63doi:10.1186/ar2434


Published:	30 May 2008

Abstract

Introduction

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.

Conclusion

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.