Aggrecanolysis in human osteoarthritis: confocal localization and biochemical characterization of ADAMTS5 - hyaluronan complexes in articular cartilages

作者：Plaas A^*; O**orn B; Yoshihara Y; Bai Y; Bloom T; Nelson F; Mikecz K; Sandy J D

来源：Osteoarthritis and Cartilage, 2007, 15(7): 719-734.

DOI：10.1016/j.joca.2006.12.008

• aggrecanase
• Osteoarthritis
• ADAMTS5
• cartilage
• hyaluronan
• confocal microscopy
• immunohistochemistry

Objective: Human osteoarthritis (OA) is characterized by aggrecanase-mediated depletion of cartilage aggrecan. We have examined the abundance, location and some biochemical properties of the six known aggrecanases (A disintegrin and metalloproteinase with thrombospondin-like motifs 1 (ADAMTS1) 4, 5, 8, 9 and 15) in normal and OA human cartilages.
Methods: Formalin-fixed, ethylenediamine tetraacetic acid (EDTA)-decalcified sections of full-depth cartilage from human OA tibial plateaus and normal control samples were studied by confocal imaging. Probes included specific antibodies to aggrecanases and two aggrecan epitopes, as well as biotinylated hyaluronan binding protein (HABP) for hyaluronan (HA) visualization. Cartilage extracts were analyzed by Western blot for the individual proteinases and aggrecan fragments.
Results: ADAMTS5 was present in association with cells throughout normal cartilage and was markedly increased in OA, particularly in clonal groups in the superficial and transitional zones, where it was predominantly co-localized with HA. Consistent with the confocal analysis, a high molecular weight Complex of ADAMTS5 and HA was isolated from human CA cartilage by isotonic salt extraction and chromatography on Superose 6. The complex eluted with an apparent molecular size of about 2 x 106 and contained major ADAMTS5 forms of 150, 60, 40 and 30 kDa. The yield of most forms on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was markedly enhanced by prior digestion of the complex with either Streptomyces hyaluronidase or chondroitinase ABC.
Conclusion: ADAMTS5 abundance and distribution in human CA cartilages is consistent with a central role for this enzyme in destructive aggrecanolysis. HA-depemdent sequestration of ADAMTS5 in the pericellular matrix may be a mechanism for regulating the activity of this proteinase in human OA cartilage.

• 出版日期2007-7