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In-vitro and in-vivo imaging of MMP activity in cartilage and joint injury.

Biochemical and biophysical research communications (2015-03-31)
Tomoaki Fukui, Elizabeth Tenborg, Jasper H N Yik, Dominik R Haudenschild
ABSTRACT

Non-destructive detection of cartilage-degrading activities represents an advance in osteoarthritis (OA) research, with implications in studies of OA pathogenesis, progression, and intervention strategies. Matrix metalloproteinases (MMPs) are principal cartilage degrading enzymes that contribute to OA pathogenesis. MMPSense750 is an in-vivo fluorimetric imaging probe with the potential to continuously and non-invasively trace real-time MMP activities, but its use in OA-related research has not been reported. Our objective is to detect and characterize the early degradation activities shortly after cartilage or joint injury with MMPSense750. We determined the appropriate concentration, assay time, and linear range using various concentrations of recombinant MMPs as standards. We then quantified MMP activity from cartilage explants subjected to either mechanical injury or inflammatory cytokine treatment in-vitro. Finally, we performed in-vivo MMP imaging of a mouse model of post-traumatic OA. Our in-vitro results showed that the optimal assay time was highly dependent on the MMP enzyme. In cartilage explant culture media, mechanical impact or cytokine treatment increased MMP activity. Injured knees of mice showed significantly higher fluorescent signal than uninjured knees. We conclude that MMPSense750 detects human MMP activities and can be used for in-vitro study with cartilage, as well as in-vivo studies of knee injury, and can offering real-time insight into the degradative processes that occurring within the joint before structural changes become evident radiographically.

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Sigma-Aldrich
Matrix Metalloproteinase-9, recombinant, >90% (SDS-PAGE), buffered aqueous solution
Sigma-Aldrich
MMP-3 human, recombinant, expressed in E. coli, ≥95% (SDS-PAGE), ≥95% (HPLC)