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Journal of orthopaedic research : official publication of the Orthopaedic Research Society2011; 29(6); 900-906; doi: 10.1002/jor.21332

Biochemical identification and immunolocalizaton of aggrecan, ADAMTS5 and inter-alpha-trypsin-inhibitor in equine degenerative suspensory ligament desmitis.

Abstract: We describe analysis of suspensory ligaments from horses with advanced degenerative suspensory ligament desmitis (DSLD) to identify the major proteoglycans (PGs), ADAMTS-aggrecanases and inter-alpha-trypsin inhibitor (IαI) components associated with ligament degeneration. Specific anatomical regions of suspensory ligaments from two normal horses and four diagnosed with DSLD were analyzed by Western blot and immunohistochemistry for the following: aggrecan, aggrecan fragments, decorin, ADAMTS4, ADAMTS5, and IαI components. When compared to normal, DSLD ligaments showed about a 15-fold increase (P < 0.0014) in aggrecan levels and markedly enhanced staining with Safranin O. The aggrecan was composed of two distinct high molecular weight core protein species. The largest species was found only in DSLD samples and it co-migrated with aggrecan synthesized by equine mesenchymal stem cells (MSC). Many of the DSLD samples also contained abnormally high concentrations of ADAMTS4, ADAMTS5, and IαI. Notably, the ADAMTS5 in DSLD samples, but not normals, was present largely as a high molecular weight complex. We conclude that ligament degeneration in DSLD is associated with matrix changes characteristic of an inflammatory nonhealing wound, specifically containing chondrogenic progenitor cells. Since aggrecan accumulation is a major feature of incomplete healing in tendon and skin of the ADAMTS5 knockout mouse, we propose that ligament failure in DSLD results from a process involving tissue inflammation and the complexation of ADAMTS5.
Copyright © 2011 Orthopaedic Research Society.
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Publication Date: 2011-01-18 PubMed ID: 21246622DOI: 10.1002/jor.21332Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study examined the suspensory ligaments in horses with degenerative suspensory ligament desmitis (DSLD), looking in particular for proteins and enzymes associated with tissue inflammation and failure to heal. The researchers found significantly increased levels of certain components, suggesting that ligament degeneration in DSLD involves ongoing inflammatory response and stalled healing processes.

Research Context

  • The study focuses on a condition in horses known as degenerative suspensory ligament desmitis (DSLD). This is a debilitating disorder that results in lameness due to degeneration of the ligaments.
  • To better understand the progression of this condition, the team analyzed components within the suspensory ligaments of horses affected, compared to those of unaffected horses.
  • The core elements examined were proteoglycans (molecules essential for connective tissue health), ADAMTS-aggrecanases (a specific group of enzymes that degrade proteoglycans), and inter-alpha-trypsin inhibitor (a protein complex related to inflammation and tissue repair).

Research Findings

  • The analysis found that the suspensory ligaments of horses with DSLD contained approximately 15 times the amount of a proteoglycan called aggrecan, compared to the ligaments in normal horses.
  • The aggrecan protein in the DSLD-affected ligaments was composed of two high molecular weight core protein species, one of which was found only in DSLD samples and seemed to match the aggrecan produced by equine mesenchymal stem cells. This suggests potential involvement of stem cells in the disease process.
  • The DSLD-affected ligaments also showed high levels of the ADAMTS4 and ADAMTS5 enzymes, as well as an elevated concentration of the inter-alpha-trypsin inhibitor.
  • The ADAMTS5 enzyme in the affected ligaments was primarily found as a large molecular weight complex, which was not the case in the normal ligaments. This finding suggests a possible role of this enzyme complex in the disease progression.

Research Implications

  • The team concluded that ligament degeneration in DSLD appears to involve an inflammatory process characteristic of nonhealing wounds, possibly involving chondrogenic progenitor cells (stem cells that can develop into cartilage).
  • The prominence of aggrecan in the disease suggests it plays a significant role in the incomplete healing processes. This is supported by the observation that accumulation of aggrecan is also a feature of the healing process in the absence of ADAMTS5, as seen in ADAMTS5 knockout mice.
  • The authors propose that the disease process in DSLD involves tissue inflammation and the binding of ADAMTS5 into a larger molecular complex. This identification and understanding of specific biochemical changes in the degenerating ligaments provide further insight into the disease and may serve as a foundation for future therapeutic approaches.

Cite This Article

APA
Plaas A, Sandy JD, Liu H, Diaz MA, Schenkman D, Magnus RP, Bolam-Bretl C, Kopesky PW, Wang VM, Galante JO. (2011). Biochemical identification and immunolocalizaton of aggrecan, ADAMTS5 and inter-alpha-trypsin-inhibitor in equine degenerative suspensory ligament desmitis. J Orthop Res, 29(6), 900-906. https://doi.org/10.1002/jor.21332

Publication

Journal of orthopaedic research : official publication of the Orthopaedic Research Society
ISSN: 1554-527X
NlmUniqueID: 8404726
Country: United States
Language: English
Volume: 29
Issue: 6
Pages: 900-906

Researcher Affiliations

Plaas, Anna
  • Department of Internal Medicine, Rush University Medical Center, Chicago, IL, USA.
Sandy, John D
    Liu, Haowen
      Diaz, Michael A
        Schenkman, Daniel
          Magnus, Robert P
            Bolam-Bretl, Courtney
              Kopesky, Paul W
                Wang, Vincent M
                  Galante, Jorge O

                    MeSH Terms

                    • ADAM Proteins / metabolism
                    • Aggrecans / metabolism
                    • Alpha-Globulins / metabolism
                    • Animals
                    • Blotting, Western
                    • Connective Tissue Diseases / metabolism
                    • Connective Tissue Diseases / pathology
                    • Endopeptidases / metabolism
                    • Female
                    • Horse Diseases / metabolism
                    • Horse Diseases / pathology
                    • Horses
                    • Immunohistochemistry
                    • Ligaments / metabolism
                    • Ligaments / pathology
                    • Male

                    Citations

                    This article has been cited 23 times.
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