In-vitro and in-vivo imaging of MMP activity in cartilage and joint injury.
In-vitro and in-vivo imaging of MMP activity in cartilage and joint injury.
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DOI：
10.1016/j.bbrc.2015.03.100
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发表时间：
2015-05-08
影响因子：
3.1
通讯作者：
Haudenschild, Dominik R.
中科院分区：
文献类型：
--
作者：
Fukui, Tomoaki;Tenborg, Elizabeth;Yik, Jasper H. N.;Haudenschild, Dominik R.
软骨降解活动的无损检测代表了骨关节炎 (OA) 研究的进步，对 OA 发病机制、进展和干预策略的研究具有重要意义。基质金属蛋白酶 (MMP) 是导致 OA 发病机制的主要软骨降解酶。 MMPSense750是一种体内荧光成像探针，具有连续、非侵入性追踪实时MMP活性的潜力，但其在OA相关研究中的应用尚未见报道。我们的目标是使用 MMPSense750 检测和表征软骨或关节损伤后不久的早期降解活动。我们使用不同浓度的重组 MMP 作为标准确定了适当的浓度、测定时间和线性范围。然后，我们量化了经过体外机械损伤或炎症细胞因子治疗的软骨外植体的 MMP 活性。最后，我们对创伤后 OA 小鼠模型进行了体内 MMP 成像。我们的体外结果表明，最佳测定时间高度依赖于 MMP 酶。在软骨外植体培养基中，机械冲击或细胞因子处理增加了 MMP 活性。受伤的小鼠膝盖比未受伤的膝盖显示出明显更高的荧光信号。我们得出的结论是，MMPSense750 可以检测人类 MMP 活性，可用于软骨的体外研究以及膝关节损伤的体内研究，并且可以在放射学上明显改变结构变化之前提供对关节内发生的降解过程的实时洞察。
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 invivo 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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影响因子：
2.3
作者：
Cattano, Nicole M.;Driban, Jeffrey B.;Sitler, Michael R.
通讯作者：
Sitler, Michael R.
影响因子：
5.6
作者：
Lozito, Thomas P.;Tuan, Rocky S.
通讯作者：
Tuan, Rocky S.
影响因子：
2.4
作者：
Barber PA;Rushforth D;Agrawal S;Tuor UI
通讯作者：
Tuor UI
DOI：
10.1016/j.bbapap.2011.06.020
发表时间：
2012-01
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Troeberg L;Nagase H
通讯作者：
Nagase H
影响因子：
7
作者：
Mapp, P. I.;Walsh, D. A.;Bowyer, J.;Maciewicz, R. A.
通讯作者：
Maciewicz, R. A.