Skip to main content
SpringerLink
Log in

The effect of administration of double stranded MicroRNA-210 on acceleration of Achilles tendon healing in a rat model

  • Original Article
  • Published:
Journal of Orthopaedic Science

Abstract

Background

Achilles tendons heal slower than other tissues, therefore requiring the developmnent of a strategy for accelerating the process. Vascular supply plays an important role in primary tendon healing, especially during the early healing phase. MicroRNA (miR)-210 has been reported as being crucial for angiogenesis, which is a key factor of tissue repair. We report herein that local injection of synthetic miR-210 into the injured Achilles tendon of a rat accelerated healing of the tendon.

Methods

Achilles tendons were transected and repaired via the Kessler suture technique in Sprague–Dawley rats. Then, double stranded (ds) miR-210 was injected into the repaired sites. The control group was injected with non-functioned dsRNA. At 2, 6 and 12 weeks, histological evaluations were performed. At two and six weeks, mechanical testing and angiogenesis were evaluated. Gene expression analysis using real-time polymerase chain reaction (PCR) and immunohistochemistry were performed at two weeks.

Result

At two and six weeks, regular dense collagen tissue in the miR-210 group was observed and the diameter of collagen fiber in the miR-210 group was significantly higher than in the control. At two weeks, the ultimate failure load was significantly higher than in the control group, and expression of VEGF, FGF2 and type I collagen was upregulated. Abundant vessels in the miR-210 group were observed at two weeks, but there was no significant difference in vessel numbers between the two groups at six weeks. At 12 weeks, repaired Achilles tendons in the miR-210 group consisted of parallel and dense fibers, whereas wavy and loose fibers were still observed in the control group.

Conclusion

The current study showed that single local injection of synthetic miR-210 promotes Achilles tendon healing in the early phase.

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
Fig. 7

Similar content being viewed by others

References

  1. Leppilahti J, Puranen J, Orava S. Incidence of Achilles tendon rupture. Acta Orthop Scand. 1996;67(3):277–9.

    Article  CAS  PubMed  Google Scholar 

  2. Inglis AE, Scott WN, Sculco TP, Patterson AH. Ruptures of the tendon Achilles: an objective assessment of surgical and non-surgical treatment. J Bone Jt Surg Am. 1976;58(7):990–3.

    CAS  Google Scholar 

  3. Persson A, Wredmark T. The treatment of total ruptures of the Achilles tendon by plaster immobilization. Int Orthop. 1979;3(2):149–52.

    CAS  PubMed  Google Scholar 

  4. Wilkins R, Bisson LJ. Operative versus nonoperative management of acute Achilles tendon ruptures. Am J Sports Med. 2012;40(9):2154–60.

    Article  PubMed  Google Scholar 

  5. Nilsson-Helander KS, Silbernagel KG, Thomee R, Faven E, Olsson N, Eriksson BI, Karlsson J. Acute Achilles tendon rupture: a randomized, controlled study comparing surgical and nonsurgical treatments using validated outcome measures. Am Sports Med. 2010;38(11):2186–93.

    Article  Google Scholar 

  6. Willits K, Amendola A, Bryant D, Mochtadi NG, Giffin JR, Fowler P, Kean CO, Kirkley A. Operative versus nonoperative treatment of acute Achilles tendon rupture: a multicenter randomized trial using accelerated functional rehabilitation. J Bone Jt Surg Am. 2010;92(17):2767–75.

    Article  Google Scholar 

  7. Zhang F, Liu H, Stile F, Lei MP, Pang Y, Oswald TM, Beck J, Dovsestt-Martin W, Lineaweaver WC. Effect of vascular endothelial growth factor on rat Achilles tendon healing. Plast Reconstr Surg. 2003;112(6):1613–9.

    Article  PubMed  Google Scholar 

  8. Huang TF, Yew TL, Chiang ER, Ma HL, Hsu CY, Hsu SH, Hsu YT, Hung SC. Mesenchymal stem cells from a hypoxic culture improve and engraft achilles tendon repair. Am J Sports Med. 2013;41(5):1117–25.

    Article  PubMed  Google Scholar 

  9. Ambros V. The functions of animal microRNAs. Nature. 2004;431(7006):350–5.

    Article  CAS  PubMed  Google Scholar 

  10. Bartel DP, Chen CZ. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116(2):281–97.

    Article  CAS  PubMed  Google Scholar 

  11. Lim LP, Lau NC, Garrett-Engele P, Grimson A, Scelter JM, Castle J, Bartel DP, Linsley PS, Johnson JM. Microarray analysis shows that some microRNAs downregulate large numbers of target mRNAs. Nature. 2005;433(7027):769–73.

    Article  CAS  PubMed  Google Scholar 

  12. Elmen J, Lindow M, Schutz S, Lawrence M, Petri A, Obad S, Lindholm M, Hedjarn M, Hanshen HF, Berger U, Gullans S, Kearney P, Sarnow P, Straarup EM, Kaupinnen S. LNA-mediated microRNA silencing in non-human primates. Nature. 2008;452(7189):896–9.

    Article  CAS  PubMed  Google Scholar 

  13. Krutzfeldt J, Rajewsky N, Braich R, Kallanthothil G, Rajeev KG, Tuschl T, Manoharan M. Silencing of microRNAs in vivo with ‘antagomirs’. Nature. 2005;438:685–9.

    Article  PubMed  Google Scholar 

  14. Nakasa T, Miyaki S, Okubo A, Hashimoto M, Nishida K, Ochi M, Asahara H. Expression of microRNA-146 in rheumatoid arthritis synovial tissue. Arthr Rheum. 2008;58(5):1284–92.

    Article  CAS  Google Scholar 

  15. Yamasaki K, Nakasa T, Miyaki S, Ishikawa M, Deie M, Adachi N, Yasunaga Y, Asahara H, Ochi M. Expression of microRNA-146a in osteoarthritis cartilage. Arthr Rheum. 2009;60(4):1035–41.

    Article  CAS  Google Scholar 

  16. Shoji T, Nakasa T, Yamasaki K, Kodama A, Miyaki S, Niimoto T, Okuhara A, Kamei N, Adachi N, Ochi M. The effect of intra-articular injection of microRNA-210 on ligament healing in a rat model. Am J Sports Med. 2012;40(11):2470–8.

    Article  PubMed  Google Scholar 

  17. Nakasa T, Ishikawa M, Shi M, Shibuya H, Adachi N, Ochi M. Acceleration of muscle regeneration by local injection of muscle specific microRNAs in rat skeletal muscle injury model. J Cell Mol Med. 2010;14(10):2495–505.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Fasanaro P, D’Alessandra Y, Di Stefano V, Malchionna R, Romani S, Pompilio G, Capogrossi MC, Martelli F. MicroRNA-210 modulates endothelial cell response to hypoxia and inhibits the receptor tyrosine kinase ligand ephrin-A3. J Biol Chem. 2008;283:15878–83.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  19. Ivan M, Harris AL, Martelli F, Kulshreshtha R. Hypoxia response and microRNAs: no longer two separate worlds. J Cell Mol Med. 2008;12(5A):1426–31.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  20. Fasanaro P, Greco S, Lorenzi M, Pescatori M, Brioschi M, Kulshereshtha R, Banfi C, Stubbs A, Calin GA, Ivan M, Capogrossi MC, Fabio. An integrated approach for experimental target identification of hypoxia-induced miR-210. J Biol Chem. 2009;284(50):35134–43.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  21. Aspenberg P, Virchenko O. Platelet concentrate injection improves Achilles tendon repair in rats. Acta Orthop Scand. 2004;75(1):93–9.

    Article  PubMed  Google Scholar 

  22. Liu SH, Yang RS, al-Shaikh R, Lane JM. Collagen in tendon, ligament, and bone healing: a current review. Clin Orthop. 1995;318:265.

    PubMed  Google Scholar 

  23. Gelberman RH, Khabie V, Cahill CJ. The revascularization of healing flexor tendons in the digital sheath: a vascular injection study in dogs. J Bone Jt Surg Am. 1991;73(6):868–81.

    CAS  Google Scholar 

  24. Sharma P, Maffulli N. Tendon injury and tendinopathy: healing and repair. J Bone Jt Surg Am. 2005;87(1):187–202.

    Article  Google Scholar 

  25. Molloy T, Wang Y, Murrel G. The role of growth factors in tendon and ligament healing. Sports Med. 2003;33(5):381–94.

    Article  PubMed  Google Scholar 

  26. Kashiwagi K, Mochizuki Y, Yasunaga Y, Ishida O, Deie M, Ochi M. Effect of transforming growth factor-beta 1 on the early stages of healing of the Achilles tendon in a rat model. Scand J Plast Reconstr Surg Hand Surg. 2004;38(4):193–7.

    Article  PubMed  Google Scholar 

  27. Virchenko O, Fahlgren A, Skoglund B, Aspenberg P. CDMP-2 injection improves early tendon healing in a rabbit model for surgical repair. Scand J Med Sci Sports. 2005;15(4):260–4.

    Article  CAS  PubMed  Google Scholar 

  28. Hu S, Huang M, Li Z, Jin F, Ghosh Z, Lijkwan MA, Fasanaro P, Sun N, Wang X, Martelli F, Robbins RC, Wu JC. MicroRNA-210 as a novel therapy for treatment of ischemic heart disease. Circulation. 2010;122(11 suppl):2124–31.

    Google Scholar 

  29. Nakasa T, Shibuya H, Nagata Y, Niimoto T, Ochi M. The inhibitory effect of microRNA-146a expression on bone destruction in collage-induced arthritis. Arthr Rheum. 2011;63(6):1582–90.

    Article  CAS  Google Scholar 

  30. Wilkins R, Bisson LJ. Operative versus nonoperative management of acute Achilles tendon ruptures. A quantitative systemic review of randomized controlled trials. Am J Sports Med. 2012;40(9):2154–60.

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This research was supported by MEXT KAKENHI Grant-in-Aid for Scientific Research (A) Grant Number 21249079 (M.O.), Scientific Research (B) Grant number 25293324 (N.K.) and Scientific Research (C) Grant Number 24592234 (T.N.).

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Orthopaedic Surgery, Integrated Health and Sciences, Institute of Biomedical and Health Science, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan

    Muhammad Andry Usman, Tomoyuki Nakasa, Takeshi Shoji, Tomohiro Kato, Yoshitaka Kawanishi, Michio Hamanishi, Naosuke Kamei & Mitsuo Ochi

  2. Department of Orthopaedic and Traumatology, Faculty of Medicine, Hasanuddin University, Jln. Perintis Kemerdekaan KM.10, Tamalanrea, Makassar, 90245, Indonesia

    Muhammad Andry Usman

Authors
  1. Muhammad Andry Usman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tomoyuki Nakasa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Takeshi Shoji
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tomohiro Kato
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yoshitaka Kawanishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Michio Hamanishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Naosuke Kamei
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Mitsuo Ochi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Muhammad Andry Usman.

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Usman, M.A., Nakasa, T., Shoji, T. et al. The effect of administration of double stranded MicroRNA-210 on acceleration of Achilles tendon healing in a rat model. J Orthop Sci 20, 538–546 (2015). https://doi.org/10.1007/s00776-015-0709-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00776-015-0709-5

Keywords

Search

Navigation