Skip to main content
SpringerLink
Log in

Calcitonin treatment is associated with less severe osteoarthritis and reduced toll-like receptor levels in a rat model

  • Original Article
  • Published:
Journal of Orthopaedic Science

Abstract

Background

Studies indicate that inflammation promotes progression of osteoarthritis. Cartilage damage is aggravated by the binding of toll-like receptors and endogenous ligands that release large amounts of cytokines and inflammation mediators. Calcitonin can inhibit degeneration of articular cartilage, by inhibiting activation of toll-like receptors and generation of endogenous ligands. To study the effect of calcitonin in the pathogenesis of osteoarthritis and the underlying molecular mechanism, we monitored levels of toll-like receptors during osteoarthritis progression, and after calcitonin injection.

Methods

Male Sprague–Dawley rats were randomly assigned to either a surgery-only or a calcitonin-treatment group (n = 35, each). To induce osteoarthritis, the anterior cruciate ligament and the medial meniscus were cut in the right knees of both groups. Rats in the calcitonin-treatment group received a subcutaneous injection of 15 IU/kg calcitonin once every other day, starting one day post-surgery, until euthanised. Signs of osteoarthritic changes were noted. The amount of collagen II was measured by antibody staining. The amounts of MMP1 and MMP3 in cartilage were measured by use of ELISA. RNA from operated and matched control knee cartilage was extracted to determine expression levels of Col2a1, ACAN, Tlr2, Tlr3, and Tlr4.

Results

Signs of osteoarthritis were less severe in rats treated with calcitonin. In the surgery-only group, Tlr2 levels increased early after surgery and then decreased substantially by the latter stages. Tlr3 levels gradually increased and correlated with the severity of osteoarthritis. Tlr4 levels were high but fluctuated over the experimental period. Calcitonin treatment was associated with lower Tlr3 and Tlr4 levels than in the surgery-only group whereas Tlr2 expression was initially lower but increased 28 days after administration of calcitonin.

Conclusion

Calcitonin treatment may lessen the severity of osteoarthritis in the rat model, perhaps by inhibition of Tlr expression in cartilage.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Santangelo KS, Pieczarka EM, Nuovo GJ, Weisbrode SE, Bertone AL. Temporal expression and tissue distribution of interleukin-1β in two strains of guinea pigs with varying propensity for spontaneous knee osteoarthritis. Osteoarthritis Cartilage. 2011;19:439–48.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  2. Kerkhof HJ, Doherty M, Arden NK, Abramson SB, Attur M, Bos SD, Cooper C, Dennison EM, Doherty SA, Evangelou E, Hart DJ, Hofman A, Javaid K, Kerna I, Kisand K, Kloppenburg M, Krasnokutsky S, Maciewicz RA, Meulenbelt I, Muir KR, Rivadeneira F, Samuels J, Sezgin M, Slagboom E, Smith AJ, Spector TD, Tamm A, Tamm A, Uitterlinden AG, Wheeler M, Zhai G, Zhang W, van Meurs JB, Valdes AM. Large-scale meta-analysis of interleukin-1 beta and interleukin-1 receptor antagonist polymorphisms on risk of radiographic hip and knee osteoarthritis and severity of knee osteoarthritis. Osteoarthritis Cartilage. 2011;19:265–71.

    Article  CAS  PubMed  Google Scholar 

  3. Farhat K, Riekenberg S, Heine H, Debarry J, Lang R, Mages J, Buwitt-Beckmann U, Röschmann K, Jung G, Wiesmüller KH, Ulmer AJ. Heterodimerization of TLR2 with TLR1 or TLR6 expands the ligand spectrum but does not lead to differential signaling. J Leukoc Biol. 2008;83:692–701.

    Article  CAS  PubMed  Google Scholar 

  4. Su SL, Tsai CD, Lee CH, Salter DM, Lee HS. Expression and regulation of Toll-like receptor 2 by IL-1b and fibronectin fragments in human articular chondrocytes. Osteoarthritis Cartilage. 2005;13:879–86.

    Article  PubMed  Google Scholar 

  5. Brentano F, Schorr O, Gay RE, Gay S, Kyburz D. RNA released from necrotic synovial fluid cells activates rheumatoid arthritis synovial fibroblasts via Toll-like receptor 3. Arthritis Rheum. 2005;52:2656–65.

    Article  CAS  PubMed  Google Scholar 

  6. Zhang Q, Hui W, Litherland GJ, Barter MJ, Davidson R, Darrah C, Donell ST, Clark IM, Cawston TE, Robinson JH, Rowan AD, Young DA. Differential Toll-like receptor-dependent collagenase expression in chondrocytes. Ann Rheum Dis. 2008;67:1633–41.

    Article  CAS  PubMed  Google Scholar 

  7. Cawston TE, Curry VA, Summers CA, Clark IM, Riley GP, Life PF, Spaull JR, Goldring MB, Koshy PJ, Rowan AD, Shingleton WD. The role of oncostatin M in animal and human connective tissue collagen turnover and its localization within the rheumatoid joint. Arthritis Rheum. 1998;41:1760–71.

    Article  CAS  PubMed  Google Scholar 

  8. Oblak A, Jerala R. Toll-like receptor 4 activation in cancer progression and therapy. Clin Dev Immunol. 2011;2011:609579.

    Article  PubMed Central  PubMed  Google Scholar 

  9. Kuroki K, Stoker AM, Sims HJ, Cook JL. Expression of Toll-like receptors 2 and 4 in stifle joint synovial tissues of dogs with or without osteoarthritis. Am J Vet Res. 2010;71:750–4.

    Article  CAS  PubMed  Google Scholar 

  10. Kim HA, Cho ML, Choi HY, Yoon CS, Jhun JY, Oh HJ, Kim HY. The catabolic pathway mediated by Toll-like receptors in human osteoarthritic chondrocytes. Arthritis Rheum. 2006;54:2152–63.

    Article  CAS  PubMed  Google Scholar 

  11. Magnusson M, Zare F, Tarkowski A. Requirement of type I interferon signaling for arthritis triggered by double-stranded RNA. Arthritis Rheum. 2006;54:148–57.

    Article  CAS  PubMed  Google Scholar 

  12. Foell D, Wittkowski H, Roth J. Mechanisms of disease: a ‘DAMP’ view of inflammatory arthritis. Nat Clin Pract Rheumatol. 2007;3:382–90.

    Article  CAS  PubMed  Google Scholar 

  13. Jin B, Sun T, Yu XH, Yang YX, Yeo AE. The effects of TLR activation on T-cell development and differentiation. Clin Dev Immunol. 2012;2012:836485.

    Article  PubMed Central  PubMed  Google Scholar 

  14. Kumar H, Kawai T, Akira S. Pathogen recognition by the innate immune system. Int Rev Immunol. 2011;30:16–34.

    Article  CAS  PubMed  Google Scholar 

  15. Chen K, Huang J, Gong W, Iribarren P, Dunlop NM, Wang JM. Toll-like receptors in inflammation, infection and cancer. Int Immunopharmacol. 2007;7:1271–85.

    Article  CAS  PubMed  Google Scholar 

  16. Zhang Z, Schluesener HJ. Mammalian toll-like receptors: from endogenous ligands to tissue regeneration. Cell Mol Life Sci. 2006;63:2901–7.

    Article  CAS  PubMed  Google Scholar 

  17. Scanzello CR, Plaas A, Crow MK. Innate immune system activation in osteoarthritis: is osteoarthritis a chronic wound? Curr Opin Rheumatol. 2008;20:565–72.

    Article  CAS  PubMed  Google Scholar 

  18. Karsdal MA, Tanko LB, Riis BJ, Sondergard BC, Henriksen K, Altman RD, Qvist P, Christiansen C. Calcitonin is involved in cartilage homeostasis: is calcitonin a treatment for OA? Osteoarthritis Cartilage. 2006;14:617–24.

    Article  CAS  PubMed  Google Scholar 

  19. Sondergaard BC, Madsen SH, Segovia-Silvestre T, Paulsen SJ, Christiansen T, Pedersen C, Bay-Jensen AC, Karsdal MA. Investigation of the direct effects of salmon calcitonin on human osteoarthritic chondrocytes. BMC Musculoskelet Disord. 2010;11:62.

    Article  PubMed Central  PubMed  Google Scholar 

  20. Chen Y, Shyu JF, Santhanagopal A, Inoue D, David JP, Dixon SJ, Horne WC, Baron R. The calcitonin receptor stimulates Shc tyrosine phosphorylation and Erk1/2 activation. Involvement of Gi, protein kinase C, and calcium. J Biol Chem. 1998;273:19809–16.

    Article  CAS  PubMed  Google Scholar 

  21. Kawai T. Akira S.TLR signaling. Cell Death Differ. 2006;13:816–25.

    Article  CAS  PubMed  Google Scholar 

  22. Karsdal MA, Byrjalsen I, Henriksen K, Riis BJ, Lau EM, Arnold M, Christiansen C. The effect of oral salmon calcitonin delivered with 5-CNAC on bone and cartilage degradation in osteoarthritic patients: a 14-day randomized study. Osteoarthritis Cartilage. 2010;18:150–9.

    Article  CAS  PubMed  Google Scholar 

  23. Hayami T, Pickarski M, Zhuo Y, Wesolowski GA, Rodan GA, le Duong T. Characterization of articular cartilage and subchondral bone changes in the rat anterior cruciate ligament transection and meniscectomized models of osteoarthritis. Bone. 2006;38:234–43.

    Article  PubMed  Google Scholar 

  24. Mankin HJ, Dorfman H, Lippiello L, Zarins A. Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips. II. Correlation of morphology with biochemical and metabolic data. J Bone Joint Surg Am. 1971;53:523–37.

    CAS  PubMed  Google Scholar 

  25. Pufe T, Petersen W, Tillmann B, Mentlein R. The splice variants VEGF121 and VEGF189 of the angiogenic peptide vascular endothelial growth factor are expressed in osteoarthritic cartilage. Arthritis Rheum. 2001;44:1082–8.

    Article  CAS  PubMed  Google Scholar 

  26. Iwaki A1, Jingushi S, Oda Y, Izumi T, Shida JI, Tsuneyoshi M, Sugioka Y. Localization and quantification of proliferating cells during rat fracture repair: detection of proliferating cell nuclear antigen by immunohistochemistry. J Bone Miner Res. 1997;12:96–102.

  27. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25:402–8.

    Article  CAS  PubMed  Google Scholar 

  28. O’Neill LA. How Toll-like receptors signal: what we know and what we don’t know. Curr Opin Immunol. 2006;18:3–9.

    Article  PubMed  Google Scholar 

  29. Yarilina A, Dicarlo E, Ivashkiv LB. Suppression of the effector phase of inflammatory arthritis by double-stranded RNA is mediated by type I IFNs. J Immunol. 2007;178(2204):2211.

    Google Scholar 

Download references

Acknowledgments

This study was supported by a grant from the Youth Science and Technology Project of Soochow Health Science and Education (KJXW2011009).

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Orthopedics, Second Affiliated Hospital of Soochow University, Suzhou, 215004, Jiangsu, China

    Jian Li, Zong-Gang Xie, Ye Xie & Qi-Rong Dong

Authors
  1. Jian Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zong-Gang Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ye Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Qi-Rong Dong
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Qi-Rong Dong.

Additional information

J. Li and Z.-G. Xie contributed equally to this work.

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, J., Xie, ZG., Xie, Y. et al. Calcitonin treatment is associated with less severe osteoarthritis and reduced toll-like receptor levels in a rat model. J Orthop Sci 19, 1019–1027 (2014). https://doi.org/10.1007/s00776-014-0629-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00776-014-0629-9

Keywords

Search

Navigation