Abstract
While overuse of the supraspinatus tendon is a leading factor in rotator cuff injury, the underlying biochemical changes have not been fully elucidated. In this study, torn human rotator cuff (supraspinatus) tendon tissue was analyzed for the presence of active cathepsin proteases with multiplex cysteine cathepsin zymography. In addition, an overuse injury to supraspinatus tendons was induced through downhill running in an established rat model. Histological analysis demonstrated that structural damage occurred by 8 weeks of overuse compared to control rats in the region of tendon insertion into bone. In both 4- and 8-week overuse groups, via zymography, there was approximately a 180% increase in cathepsin L activity at the insertion region compared to the controls, while no difference was found in the midsubstance area. Additionally, an over 400% increase in cathepsin K activity was observed for the insertion region of the 4-week overused tendons. More cathepsin K and L immunostaining was observed at the insertion region of the overuse groups compared to controls. These results provide important information on a yet unexplored mechanism for tendon degeneration that may operate alone or in conjunction with other proteases to contribute to chronic tendinopathy.
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Abbreviations
- MMP:
-
Matrix metalloproteinase
- OCT:
-
Optimum cutting temperature
- H&E:
-
Hematoxylin and eosin
- SDS:
-
Sodium dodecyl sulfate
References
Archambault, J. M., S. A. Jelinsky, S. P. Lake, et al. Rat supraspinatus tendon expresses cartilage markers with overuse. J. Orthop. Res. 25(5):617–624, 2007.
Attia, M., A. Scott, A. Duchesnay, et al. Alterations of overused supraspinatus tendon: a possible role of glycosaminoglycans and HARP/pleiotrophin in early tendon pathology. J. Orthop. Res. 30(1):61–71, 2011.
Attia, M., E. Huet, C. Gossard, et al. Early events of overused supraspinatus tendons involve matrix metalloproteinases and EMMPRIN/CD147 in the absence of inflammation. Am. J. Sports Med. 41(4):908–917, 2013.
Barbato, J. C., L. G. Koch, A. Darvish, et al. Spectrum of aerobic endurance running performance in eleven inbred strains of rats. J. Appl. Physiol. 85(2):530–536, 1998.
Benjamin, M., H. Toumi, J. R. Ralphs, et al. Where tendons and ligaments meet bone: attachment sites (‘entheses’) in relation to exercise and/or mechanical load. J. Anat. 208(4):471–490, 2006.
Berglund, M. E., D. A. Hart, C. Reno, and M. Wiig. Growth factor and protease expression during different phases of healing after rabbit deep flexor tendon repair. J. Orthop. Res. 29(6):886–892, 2011.
Blevins, F. T. Rotator cuff pathology in athletes. Sports Med. 24(3):205–220, 1997.
Boileau, P., N. Brassart, D. J. Watkinson, et al. Arthroscopic repair of full-thickness tears of the supraspinatus: does the tendon really heal? J. Bone Joint Surg. Am. 87(6):1229–1240, 2005.
Bromme, D., and F. Lecaille. Cathepsin K inhibitors for osteoporosis and potential off-target effects. Expert Opin. Investig. Drugs 18(5):585–600, 2009.
Bromme, D., Z. Li, M. Barnes, and E. Mehler. Human cathepsin V functional expression, tissue distribution, electrostatic surface potential, enzymatic characterization, and chromosomal localization. Biochemistry 38(8):2377–2385, 1999.
Cook, J. L., J. A. Feller, S. F. Bonar, and K. M. Khan. Abnormal tenocyte morphology is more prevalent than collagen disruption in asymptomatic athletes’ patellar tendons. J. Orthop. Res. 22(2):334–338, 2004.
Cunnane, G., O. FitzGerald, K. M. Hummel, et al. Collagenase, cathepsin B and cathepsin L gene expression in the synovial membrane of patients with early inflammatory arthritis. Rheumatology 38(1):34–42, 1999.
Dejica, V. M., J. S. Mort, S. Laverty, et al. Cleavage of type II collagen by cathepsin K in human osteoarthritic cartilage. Am. J. Pathol. 173(1):161–169, 2008.
Deval, C., S. Mordier, C. Obled, et al. Identification of cathepsin L as a differentially expressed message associated with skeletal muscle wasting. Biochem. J. 360(Pt 1):143, 2001.
Eeckhout, Y., and G. Vaes. Further studies on the activation of procollagenase, the latent precursor of bone collagenase. Effects of lysosomal cathepsin B, plasmin and kallikrein, and spontaneous activation. Biochem. J. 166(1):21–31, 1977.
Fu, S. C., B. P. Chan, W. Wang, et al. Increased expression of matrix metalloproteinase 1 (MMP1) in 11 patients with patellar tendinosis. Acta Orthop. Scand. 73(6):658–662, 2002.
Garnero, P., O. Borel, I. Byrjalsen, et al. The collagenolytic activity of cathepsin K is unique among mammalian proteinases. J. Biol. Chem. 273(48):32347–32352, 1998.
Gimbel, J. A., J. P. Van Kleunen, S. Mehta, et al. Supraspinatus tendon organizational and mechanical properties in a chronic rotator cuff tear animal model. J. Biomech. 37(5):739–749, 2004.
Goretzki, L., M. Schmitt, K. Mann, et al. Effective activation of the proenzyme form of the urokinase-type plasminogen activator (pro-uPA) by the cysteine protease cathepsin L. FEBS Lett. 297(1–2):112–118, 1992.
Gotoh, M., K. Hamada, H. Yamakawa, et al. Significance of granulation tissue in torn supraspinatus insertions: an immunohistochemical study with antibodies against interleukin-1β, cathepsin D, and matrix metalloprotease-1. J. Orthop. Res. 15(1):33–39, 1997.
Joshi, S. K., H. T. Kim, B. T. Feeley, and X. Liu. Differential ubiquitin-proteasome and autophagy signaling following rotator cuff tears and suprascapular nerve injury. J. Orthop. Res. 32(1):138–144, 2014.
Kannus, P., and L. Jozsa. Histopathological changes preceding spontaneous rupture of a tendon. A controlled study of 891 patients. J. Bone Joint Surg. Am. 73(10):1507–1525, 1991.
Kirschke, H., B. Wiederanders, D. Bromme, and A. Rinne. Cathepsin S from bovine spleen. Purification, distribution, intracellular localization and action on proteins. Biochem. J. 264(2):467–473, 1989.
Kozawa, E., Y. Nishida, X. W. Cheng, et al. Osteoarthritic change is delayed in a Ctsk-knockout mouse model of osteoarthritis. Arthritis Rheum. 64(2):454–464, 2012.
Li, W. A., Z. T. Barry, J. D. Cohen, et al. Detection of femtomole quantities of mature cathepsin K with zymography. Anal. Biochem. 401(1):91–98, 2010.
Lo, I. K. Y., L. L. Marchuk, R. Hollinshead, et al. Matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase mRNA levels are specifically altered in torn rotator cuff tendons. Am. J. Sport Med. 32(5):1223–1229, 2004.
Maffulli, N., U. G. Longo, F. Franceschi, et al. Movin and Bonar scores assess the same characteristics of tendon histology. Clin. Orthop. Relat. Res. 466(7):1605–1611, 2008.
Maganaris, C. N., M. V. Narici, L. C. Almekinders, and N. Maffulli. Biomechanics and pathophysiology of overuse tendon injuries: ideas on insertional tendinopathy. Sports Med. 34(14):1005–1017, 2004.
Masarachia, P. J., B. L. Pennypacker, M. Pickarski, et al. Odanacatib reduces bone turnover and increases bone mass in the lumbar spine of skeletally mature ovariectomized rhesus monkeys. J. Bone Miner. Res. 27(3):509–523, 2012.
Millar, N. L., A. Q. Wei, T. J. Molloy, et al. Heat shock protein and apoptosis in supraspinatus tendinopathy. Clin. Orthop. Relat. Res. 466(7):1569–1576, 2008.
Morko, J. P., M. Soderstrom, A. M. Saamanen, et al. Up regulation of cathepsin K expression in articular chondrocytes in a transgenic mouse model for osteoarthritis. Ann. Rheum. Dis. 63(6):649–655, 2004.
Morko, J., R. Kiviranta, K. Joronen, et al. Spontaneous development of synovitis and cartilage degeneration in transgenic mice overexpressing cathepsin K. Arthritis Rheum. 52(12):3713–3717, 2005.
Oliva, F., D. Barisani, A. Grasso, and N. Maffulli. Gene expression analysis in calcific tendinopathy of the rotator cuff. Eur. Cells Mater. 21:548–557, 2011.
Park, K. Y., W. A. Li, and M. O. Platt. Patient specific proteolytic activity of monocyte-derived macrophages and osteoclasts predicted with temporal kinase activation states during differentiation. Integr. Biol. (Camb.) 4(12):1459–1469, 2012.
Patterson-Kane, J. C., A. M. Wilson, E. C. Firth, et al. Exercise-related alterations in crimp morphology in the central regions of superficial digital flexor tendons from young thoroughbreds: a controlled study. Equine Vet. J. 30(1):61–64, 1998.
Riley, G. P., R. L. Harrall, C. R. Constant, et al. Tendon degeneration and chronic shoulder pain: changes in the collagen composition of the human rotator cuff tendons in rotator cuff tendinitis. Ann. Rheum. Dis. 53(6):359–366, 1994.
Scott, A., J. L. Cook, D. A. Hart, et al. Tenocyte responses to mechanical loading in vivo: a role for local insulin-like growth factor 1 signaling in early tendinosis in rats. Arthritis Rheum. 56(3):871–881, 2007.
Sharma, P., and N. Maffulli. Tendon injury and tendinopathy: healing and repair. J. Bone Joint Surg. Am. 87(1):187–202, 2005.
Sher, J. S. Anatomy, biomechanics, and pathophysiology of rotator cuff disease. In: Disorders of the Shoulder: Diagnosis and Management, edited by J. P. Iannotti, and G. R. Williams. Philadelphia: Lippincott, Williams, and Wilkins, 1999, pp. 3–29.
Silverstein, B., E. Welp, N. Nelson, and J. Kalat. Claims incidence of work-related disorders of the upper extremities: washington state, 1987 through 1995. Am. J. Public Health 88(12):1827–1833, 1998.
Soslowsky, L. J., J. E. Carpenter, C. M. DeBano, et al. Development and use of an animal model for investigations on rotator cuff disease. J. Shoulder Elb. Surg. 5(5):383–392, 1996.
Soslowsky, L. J., S. Thomopoulos, S. Tun, et al. Neer award 1999 Overuse activity injures the supraspinatus tendon in an animal model: a histologic and biomechanical study. J. Shoulder Elb. Surg. 9(2):79–84, 2000.
Wilder, C. L., K. Y. Park, P. M. Keegan, and M. O. Platt. Manipulating substrate and pH in zymography protocols selectively distinguishes cathepsins K, L, S, and V activity in cells and tissues. Arch. Biochem. Biophys. 516(1):52–57, 2011.
Acknowledgments
The authors thank Jennifer Lei for assistance in animal studies, Meredith Fay and Ang (Kevin) Li for help in tissue processing, and Bernard Kippelen’s laboratory at Georgia Tech for use of the circular polarized microscope. This study was supported by a National Football League Charities Medical Grant, a Regenerative Engineering and Medicine Seed Grant (REM) from Georgia Tech and Emory University through the Atlanta Clinical & Translational Science Institute (Advancing Translational Sciences of the National Institutes of Health, UL1TR000454) and the National Institute of Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of Health under Award Number R01AR063692. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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Associate Editor Michael S. Detamore oversaw the review of this article.
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Seto, S.P., Parks, A.N., Qiu, Y. et al. Cathepsins in Rotator Cuff Tendinopathy: Identification in Human Chronic Tears and Temporal Induction in a Rat Model. Ann Biomed Eng 43, 2036–2046 (2015). https://doi.org/10.1007/s10439-014-1245-8
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DOI: https://doi.org/10.1007/s10439-014-1245-8