MMP and non-MMP-mediated release of aggrecan and its fragments from articular cartilage: a comparative study of three different aggrecan and glycosaminoglycan assays

Abstract

The stimulation of bovine articular cartilage explants with OSM/TNFalpha released aggrecan fragments both in an MMP and non-MMP-mediated route. These immunoassays carry a potential as diagnostic tools for the quantitative assessment of the cartilage turnover in RA patients in addition to their utility in ex vivo explant cultures.

Biglycan and Fibromodulin Have Essential Roles in Regulating Chondrogenesis and Extracellular Matrix Turnover in Temporomandibular Joint Osteoarthritis

“…30 Two monoclonal antibodies were used in a sandwich ELISA, including antibody AF-28 31 recognizing 342 FFGVG-G2 fragment and antibody, F78 30 recognizing epitopes within the G1 and G2 globular domains of aggrecan.…”

“…30 To determine whether overactive TGF-␤1 signaling affects MMP-mediated aggrecan degradation, we measured the amount of aggrecan neo-epitopes released in the culture medium of the explants cultures using an ELISA based on a monoclonal antibody targeting FFGVG sequences. 42 The release of this aggrecan fragment in the Bgn Ϫ/0 Fmod Ϫ/Ϫ explant culture medium was not significantly different from wild-type explant culture medium ( Figure 6B).…”

Matrix metalloproteinase and aggrecanase generated aggrecan fragments: implications for the diagnostics and therapeutics of destructive joint diseases

“…Recently, we have also developed immunoassays for detection of MMP-( 342 FFGVG) or aggrecanase-generated ( 374 ARGSVI) aggrecan fragments. We were able to detect aggrecanase-generated fragments in the initial phase (day 2-7) and MMP analytes in the late phase (after day 12) of the degradation of bovine articular cartilage explants subjected to the catabolic effect of OSM/TNFa Sumer et al, 2007]. The study by Fosang et al [2000], however, demonstrates the Fig.…”

“…Several investigators have reported immunoassays for the detection and quantification of neo-epitopes of aggrecan Fosang et al, 1995;Sumer et al, 2007]. Pratta et al [2006] detected aggrecanasegenerated 374 ARGSVI molecules using a sandwich ELISA, in which they combined an anti-keratan sulphate antibody (as a catcher) with an anti 374 ARGS-VI monoclonal antibody used for detection of the captured fragments .…”

Proteoglycomics: Recent Progress and Future Challenges

“…In addition, mAbs raised against neo-epitopes generated by PG proteolysis have been extensively used for studying the distribution and catabolism of connective tissue PGs in normal and arthritic cartilage (Caterson et al, 2000;Fosang et al, 1995;Karsdal et al, 2008;Smith et al, 2009;Sumer et al, 2007). Lauer et al (2007) employed fluorophore-assisted carbohydrate electrophoresis (FACE) to examine the changes in the kidney HS disaccharide composition at different time points from the onset of streptozotocin-induced diabetes in rats.…”