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The adult human anterior cruciate ligament (ACL) has poor functional healing response. Transforming growth factor (TGF)-ββ1 enhances the wound repair by stimulating matrix proteins deposition as well as the proliferation and migration of cells. However, the function of the TGF-ββ1-induced matrix metalloproteinases' (MMPs) activities in the wound healing process is poorly understood. In this study, exogenous MMP-2 is added to mimic the TGF-ββ1-induced MMP-2 expression. Role of NF-κκB pathway is further examined. Our results show that TGF-ββ1 induces dramatic elevation of MMP-2 activities and the MMP-2//tissue inhibitors of metalloproteinases ratio. Furthermore, the exogenous MMP-2 significantly promoted in vitro wound healing abilities of ACL fibroblasts that are significantly blocked with the addition of its inhibitors. TGF-ββ1 also increases the proliferation of ACL fibroblasts whereas MMP-2 alone does not, indicating that MMP-2 activities are not involved in the proliferation. TGF-ββ1-induced MMP-2 activity is inhibited by Bay11-7082 and Bay11-7085 (NF-κκB inhibitors). Our results demonstrate that increased TGF-ββ1 facilitates the ACL healing process by promoting the fibroblasts migration and proliferation. The migration process is mediated by MMP-2 and NF-κκB pathway is involved in TGF-ββ1-mediated MMP-2 release. [ABSTRACT FROM AUTHOR]

Anterior cruciate ligament (ACL) has poor healing ability and an injured ACL would induce the degeneration of other intra-articular connective tissues. However, the coordinated expression and activities of matrix metalloproteinase (MMPs) in intra-articular tissues induced by ACL rupture were poorly understood. With a rat ACL rotating injury model, we found that after ACL injury, the mRNA levels of MMP-13, TIMP-1, and CD147 were significantly elevated in ACL, posterior cruciate ligament (PCL), synovium, meniscus, and cartilage. Also, MMP-2 activity was also elevated significantly in a time-dependent manner in all intra-articular tissues. Synovium showed the most capability to release MMPs, whereas ACL showed the highest MMP-13/TIMP-1 ratio. Generic MMP activity assay and zymography showed time dependent elevation of MMP activities in synovial fluids (SF). We concluded that the ACL injury would induce a coordinated response of intra-articular tissues to express MMPs, TIMPs, and CD147. The MMP activities in the microenvironment in SF would accumulate, released by all the intra-articular tissues, which would contribute to the knee damage and degeneration induced by ACL injury. [ABSTRACT FROM AUTHOR]

The anterior cruciate ligament (ACL) is known to have a poor healing ability, especially in comparison with the medial collateral ligament, which can heal relatively well. In this study, we detected significant increases in the mRNA levels of multiple matrix metalloproteinases (MMPs) (MMP-1, -2, -7, -9, -11, -14, -17, -21, -23A, -24, -25, -27, and -28) and tissue inhibitors of metalloproteinases (TIMPs) (TIMP-1, -2, -3, and -4) in ACL fibroblasts after an in vitro injury with an equi-biaxial stretch chamber. However, only some MMPs (MMP-7, -9, -14, -21, and -24) showed increases in injured medial collateral ligament fibroblasts, and to a much lesser degree than that observed in the injured ACL fibroblasts. Zymography revealed a 6.3-fold increase of MMP-2 activity in injured ACL but not medial collateral ligament fibroblasts, which agrees with the global MMP activities assay. Bay-11 and curcumin can significantly decrease MMP-2 activities to 13% and 29% in injured ACL fibroblasts, respectively, which implies the involvement of p65 subunits of nuclear factor κB and AP-1 pathways. Furthermore, Bay-11 can decrease the global MMP activity released from injured ACL fibroblasts in a dose-dependent manner. In summary, the differential expression and activities of MMPs might help to explain the poor healing ability of ACL, and the p65 subunit of nuclear factor κB might be a potential target to facilitate the ACL repair. [ABSTRACT FROM AUTHOR]

Inflammation and accumulation of matrix metalloproteinases (MMPs) in synovial fluids may be involved in the poor healing ability of the anterior cruciate ligament (ACL) after injury. With a rat ACL rotating injury model, we found that levels of IL-1β, IL-6, and TNF-α were significantly higher in synovial fluids after ACL injury. MMP-2 activity and global MMP activity in synovial fluids also increased significantly in a time-dependent manner. Ex vivo studies showed that all tissues contributed to the elevation of MMP-2 in synovial fluids, especially synovium and the injured ACL. We concluded that although the regular wound-healing mechanism also occurs after ACL injury, accumulation of MMP activity in the synovial fluids, due to all of the intraarticular tissues, may be at least one of the important reasons why an injured ACL cannot be repaired. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:243-248, 2009 [ABSTRACT FROM AUTHOR]

The human ACL (anterior cruciate ligament) is susceptible to injury but has poor healing response, whereas an injured MCL (medial collateral ligament) can be repaired relatively well. Since MMPs (matrix metalloproteases) and TIMPs (tissue inhibitor of metalloproteases) are involved in this tissue remodeling process, investigation of different response of MMPs and TIMPs family in ACL and MCL fibroblasts might lead to understanding the differential matrix remodeling process as well as their different healing ability. The first step would be determination of whether these tissue remodeling effectors are present in ligaments. In this study, we designed primers for real-time RT-PCR and determined the expression of MMPs and TIMPs family in ACL and MCL fibroblasts with synovium as a positive control. Semiquantitative RT-PCR revealed that multiple MMPs and TIMPs expressed in human ACL and MCL fibroblasts except MMP-8, 10, 12, 13, 15, 16, 20, and 26. MMP-7 was present in MCL but not in ACL fibroblast. Quantitative real-time RT-PCR showed that mRNA levels of MMP-1, 2, 14, 17, 23A, and 23B and TIMP-4 are significantly higher in MCL than in ACL fibroblasts. However, MMP-3 is higher in ACL than in MCL fibroblasts. We conclude that numerous MMPs and TIMPs family members that are differentially expressed in ACL and MCL might be involved in the differential matrix remodeling process as well as the differential healing ability of ACL and MCL. [ABSTRACT FROM AUTHOR]