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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 canmore » offering real-time insight into the degradative processes that occurring within the joint before structural changes become evident radiographically. - Highlights: • MMPSense750 is near-infrared fluorescent probe which can detect MMP activity. • MMPSense750 can detect human MMP-3, -9, and -13. • The reaction kinetics with MMPSense750 were different for the three MMPs. • MMPSense750 can visualized real time MMP activity in mouse injured knees. • MMPSense750 is convenient tool to evaluate real-time MMP activity non-invasively.« less

Authors:
; ; ;
Publication Date:
OSTI Identifier:
22462041
Resource Type:
Journal Article
Journal Name:
Biochemical and Biophysical Research Communications
Additional Journal Information:
Journal Volume: 460; Journal Issue: 3; Other Information: Copyright (c) 2015 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; BIOMEDICAL RADIOGRAPHY; BONE JOINTS; CARTILAGE; CONCENTRATION RATIO; CULTURE MEDIA; ENZYMES; FLUORESCENCE; HUMAN POPULATIONS; IN VITRO; IN VIVO; INFLAMMATION; INJURIES; MICE; PATHOGENESIS; PROBES; SIGNALS

Citation Formats

Fukui, Tomoaki, Tenborg, Elizabeth, Yik, Jasper H.N., and Haudenschild, Dominik R., E-mail: DRHaudenschild@ucdavis.edu. In-vitro and in-vivo imaging of MMP activity in cartilage and joint injury. United States: N. p., 2015. Web. doi:10.1016/J.BBRC.2015.03.100.
Fukui, Tomoaki, Tenborg, Elizabeth, Yik, Jasper H.N., & Haudenschild, Dominik R., E-mail: DRHaudenschild@ucdavis.edu. In-vitro and in-vivo imaging of MMP activity in cartilage and joint injury. United States. doi:10.1016/J.BBRC.2015.03.100.
Fukui, Tomoaki, Tenborg, Elizabeth, Yik, Jasper H.N., and Haudenschild, Dominik R., E-mail: DRHaudenschild@ucdavis.edu. Fri . "In-vitro and in-vivo imaging of MMP activity in cartilage and joint injury". United States. doi:10.1016/J.BBRC.2015.03.100.
@article{osti_22462041,
title = {In-vitro and in-vivo imaging of MMP activity in cartilage and joint injury},
author = {Fukui, Tomoaki and Tenborg, Elizabeth and Yik, Jasper H.N. and Haudenschild, Dominik R., E-mail: DRHaudenschild@ucdavis.edu},
abstractNote = {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. - Highlights: • MMPSense750 is near-infrared fluorescent probe which can detect MMP activity. • MMPSense750 can detect human MMP-3, -9, and -13. • The reaction kinetics with MMPSense750 were different for the three MMPs. • MMPSense750 can visualized real time MMP activity in mouse injured knees. • MMPSense750 is convenient tool to evaluate real-time MMP activity non-invasively.},
doi = {10.1016/J.BBRC.2015.03.100},
journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 3,
volume = 460,
place = {United States},
year = {2015},
month = {5}
}