title: In-vitro and in-vivo imaging of MMP activity in cartilage and joint injury.
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.
authors.affiliation: Lawrence J. Ellison Musculoskeletal Research Center, Department of Orthopaedic Surgery, University of California Davis Medical Center, 4635 Second Avenue Suite 2000, Sacramento CA 95817, USA.
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Fukui
authors.affiliation: Lawrence J. Ellison Musculoskeletal Research Center, Department of Orthopaedic Surgery, University of California Davis Medical Center, 4635 Second Avenue Suite 2000, Sacramento CA 95817, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Tenborg
authors.affiliation: Lawrence J. Ellison Musculoskeletal Research Center, Department of Orthopaedic Surgery, University of California Davis Medical Center, 4635 Second Avenue Suite 2000, Sacramento CA 95817, USA.
authors.forename: Jasper H N
authors.initials: JH
authors.lastname: Yik
authors.affiliation: Lawrence J. Ellison Musculoskeletal Research Center, Department of Orthopaedic Surgery, University of California Davis Medical Center, 4635 Second Avenue Suite 2000, Sacramento CA 95817, USA. Electronic address: DRHaudenschild@ucdavis.edu.
authors.forename: Dominik R
authors.initials: DR
authors.lastname: Haudenschild
mesh_terms: D000818:Animals; D002356:Cartilage / Q000201:enzymology*; D066298:In Vitro Techniques; D007718:Knee Injuries / Q000201:enzymology*; D008297:Male; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D020782:Matrix Metalloproteinases
keywords: Cartilage; In-vivo imaging; MMP activity; MMPSense750; Osteoarthritis
references.citation: Osteoarthritis Cartilage. 2013 Jan;21(1):151-6
references.pmid: 23025928
references.citation: BMC Neurosci. 2012;13:76
references.pmid: 22742423
references.citation: Arthritis Rheum. 2000 Apr;43(4):852-8
references.pmid: 10765930
references.citation: Osteoarthritis Cartilage. 1998 Mar;6(2):115-24
references.pmid: 9692066
references.citation: FASEB J. 1998 Sep;12(12):1075-95
references.pmid: 9737711
references.citation: Med Clin North Am. 2009 Jan;93(1):101-26, xi
references.pmid: 19059024
references.citation: Osteoarthritis Cartilage. 2010 Apr;18(4):593-600
references.pmid: 20067755
references.citation: Contrast Media Mol Imaging. 2013 Jan-Feb;8(1):1-11
references.pmid: 23109387
references.citation: Osteoarthritis Cartilage. 2013 Jan;21(1):209-16
references.pmid: 23069857
references.citation: Osteoarthritis Cartilage. 2013 Mar;21(3):470-80
references.pmid: 23257246
references.citation: J Periodontal Res. 2013 Jun;48(3):357-66
references.pmid: 23058002
references.citation: Arthritis Care Res (Hoboken). 2013 May;65(5):703-11
references.pmid: 23203864
references.citation: Arthritis Res Ther. 2013;15(6):R223
references.pmid: 24373218
references.citation: Pharm Res. 2014 Jun;31(6):1450-9
references.pmid: 23619595
references.citation: Arthritis Res Ther. 2014;16(1):R11
references.pmid: 24428864
references.citation: J Clin Densitom. 2015 Apr-Jun;18(2):239-44
references.pmid: 25304911
references.citation: Hum Genomics. 2010 Feb;4(3):194-201
references.pmid: 20368140
references.citation: J Cell Biochem. 2011 Dec;112(12):3507-14
references.pmid: 21815191
references.citation: J Cell Physiol. 2012 Feb;227(2):534-49
references.pmid: 21437907
references.citation: Biochim Biophys Acta. 2012 Jan;1824(1):133-45
references.pmid: 21777704
references.citation: Mol Imaging Biol. 2012 Feb;14(1):32-9
references.pmid: 21384207
references.citation: BMC Musculoskelet Disord. 2011;12:273
references.pmid: 22122951
references.citation: Arthritis Res Ther. 2011;13(5):R142
references.pmid: 21888631
references.citation: Arthritis Rheum. 2012 Jun;64(6):1697-707
references.pmid: 22392533
references.citation: Osteoarthritis Cartilage. 2012 Jul;20(7):773-82
references.pmid: 22531459
references.citation: Arthritis Rheum. 2012 Nov;64(11):3583-93
references.pmid: 22729466