Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Local NO synthase inhibition produces histological and functional recovery in Achilles tendon of rats after tenotomy

Tendon repair and local NOS inhibition

Abstract

Repair of injured tendon is a very slow process and involves the release of many molecules, including nitric oxide. We investigate the influence of local nitrergic inhibition in histological and functional recovery of injured Achilles tendon. A standard murine model of tendon injury by rupture was used. The animals were divided into three experimental groups: control, injury + vehicle (normal saline) and injury + Nω-nitro-L-arginine methyl ester (L-NAME). The products were injected into the paratendinous region every 2 days and body weight gain and Achilles functional index (AFI) were evaluated on days 0, 7, 14 and 21 after tendon injury. On day 21 post-injury, the animals were killed to evaluate nitric oxide production and tissue organization. We observed that tendon surgical division led to increased tissue nitrite levels, which were reduced in L-NAME-treated rats. The AFI revealed functional recovery of L-NAME-treated animals on day 21 post-injury, which was not observed in the saline-treated group. Microscopic analysis of hematoxylin-eosin staining and collagen autofluorescence showed that L-NAME-treated rats had more aligned areas of collagen fibers and that the diameter of newly organized collagen in this group was also greater than that in the vehicle-treated one. We demonstrate that local treatment with L-NAME significantly improves the functional parameters and accelerates histomorphological recovery.

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

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

References

  1. Abate M, Gravare SK, Siljeholm C, Di Iorio A, De Amicis D, Salini V, Werner S, Paganelli R (2009) Pathogenesis of tendinopathies: inflammation or degeneration? Arthritis Res Ther 11:235

  2. Ackermann PW, Li J, Lundeberg T, Kreicbergs A (2003) Neuronal plasticity in relation to nociception and healing of rat Achilles tendon. J Orthop Res 21:432–441

  3. Barmakian JT, Lin H, Green SM, Posner MA, Casar RS (1994) Comparison of a suture technique with the modified Kessler method: resistance to gap formation. J Hand Surg 19A:777–781

  4. Bogdan C, Röllinghoff M, Diefenbach A (2000) Reactive oxygen and reactive nitrogen intermediates in innate and specific immunity. Curr Opin Immunol 12:64–76

  5. Bokhari AR, Murrell GAC (2012) The role of nitric oxide in tendon healing. J Shoulder Elbow Surg 21:238–244

  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

  7. Bring DKI, Kreicbergs A, Renstrom PAFH, Ackermann PW (2007) Physical activity modulates nerve plasticity and stimulates repair after Achilles tendon rupture. J Orthop Res 25:164–172

  8. Bruns J, Kampen J, Kahrs J, Plitz W (2000) Achilles tendon rupture: experimental results on spontaneous repair in a sheep-model. Knee Surg Sports Traumatol Arthrosc 8:364–369

  9. Cao M, Westerhausen-Larson A, Niyibizi C, Kavalkovich K, Georgescu HI, Rizzo CF, Hebda PA, Stefanovic-Racic M, Evans CH (1997) Nitric oxide inhibits the synthesis of type-II collagen without altering Col2A1 mRNA abundance: prolyl hydroxylase as a possible target. Biochem J 324:305–310

  10. Cho NS, Hwang JH, Lee YT, Chae SW (2011) Tendinosis-like histologic and molecular changes of the Achilles tendon to repetitive stress. Clin Orthop Relat Res 469:3172–3180

  11. Cook JL, Feller JA, Bonar SF, Khan KM (2004) Abnormal tenocyte morphology is more prevalent than collagen disruption in asymptomatic athletes’ patellar tendons. J Orthop Res 22:334–338

  12. Dahlgren LA, Mohammed HO, Nixon AJ (2005) Temporal expression of growth factors and matrix molecules in healing tendon lesions. J Orthop Res 23:84–92

  13. Darmani H, Crossan JC, Curtis A (2004) Single dose of inducible nitric oxide synthase inhibitor induces prolonged inflammatory cell accumulation and fibrosis around injured tendon and synovium. Mediators Inflamm 13:157–164

  14. Dooley A, Gao B, Shi-Wen X, Abraham DJ, Black CM, Jacobs M, Bruckdorfer KR (2007) Effect of nitric oxide and peroxynitrite on type I collagen synthesis in normal and scleroderma dermal fibroblasts. Free Radic Biol Med 43:253–264

  15. Ferrini MG, Vernet D, Magee TR, Shahed A, Qian A, Rajfer J, Gonzalez-Cadavid NF (2002) Antifibrotic role of inducible nitric oxide synthase. Nitric Oxide 6:283–294

  16. Gigante A, Cesari E, Busilacchi A, Manzotti S, Kyriakidou K, Greco F, Primio RD, Mattioli-Belmonte M (2009) Collagen I membranes for tendon repair: effect of collagen fiber orientation on cell behavior. J Orthop Res 27:826–832

  17. Hoell T, Huschak G, Beier A, Hüttmann G, Minkus Y, Holzhausen H, Meisel H (2006) Auto fluorescence of intervertebral disc tissue: a new diagnostic tool. Eur Spine J 15:345–353

  18. Lin J-H, Wang M-X, Wei A, Zhu W, Diwan AD, Murrell GAC (2001a) Temporal expression of nitric oxide synthase isoforms in healing Achilles tendon. J Orthop Res 19:136–142

  19. Lin J, Wang M-X, Wei A, Zhu W, Murrell GAC (2001b) The cell specific temporal expression of nitric oxide synthase isoforms during Achilles tendon healing. Inflamm Res 50:515–522

  20. Longo UG, Ronga M, Maffulli N (2009) Achilles tendinopathy. Sports Med Arthrosc 17:112–126

  21. Maffulli N, Longo U, Franceschi F, Rabitti C, Denaro V (2008) Movin and Bonar scores assess the same characteristics of tendon histology. Clin Orthop Relat Res 466:1605–1611

  22. Molloy TJ, Wang Y, Horner A, Skerry TM, Murrell GAC (2006) Microarray analysis of healing rat Achilles tendon: evidence for glutamate signaling mechanisms and embryonic gene expression in healing tendon tissue. J Orthop Res 24:842–855

  23. Moncada S, Higgs A (1995) Molecular mechanisms and therapeutic strategies related to nitric oxide. FASEB J 9:1319–1330

  24. Moncada S, Palmer RM, Higgs EA (1991) Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev 43:109–142

  25. Murrell GAC (2007) Oxygen free radicals and tendon healing. J Shoulder Elbow Surg 16:S208–S214

  26. Murrell GAC, Lilly EG, Davies H, Best TM, Goldner RD, Seaber AV (1992) The Achilles functional index. J Orthop Res 10:398–404

  27. Murrell GAC, Szabo C, Hannafin JA, Jang D, Dolan MM, Deng X-H, Murrell DF, Warren RF (1997) Modulation of tendon healing by nitric oxide. Inflamm Res 46:19–27

  28. Murrell G, Tang G, Appleyard R, del Soldato P, Wang M-X (2008) Addition of nitric oxide through nitric oxide-paracetamol enhances healing rat Achilles tendon. Clin Orthop Relat Res 466:1618–1624

  29. Padanilam TG (2009) Chronic Achilles tendon ruptures. Foot Ankle Clin N Am 14:711–728

  30. Riley G (2008) Tendinopathy - from basic science to treatment. Nat Clin Pract Rheumatol 4:82–89

  31. Schepsis AA, Jones H, Haas AL (2002) Achilles tendon disorders in athletes. Am J Sports Med 30:287–305

  32. Scutt N, Rolf CG, Scutt A (2006) Glucocorticoids inhibit tenocyte proliferation and tendon progenitor cell recruitment. J Orthop Res 24:173–182

  33. Sharma P, Maffulli N (2006) Biology of tendon injury: healing, modeling and remodeling. J Musculoskelet Neuronal Interact 6:181–190

  34. Stamler JS, Meissner G (2001) Physiology of nitric oxide in skeletal muscle. Physiol Rev 81:209–237

  35. Strom AC, Casillas MM (2009) Achilles tendon rehabilitation. Foot Ankle Clin N Am 14:773–782

  36. Szomor ZL, Appleyard RC, Murrel GAC (2006) Overexpression of nitric oxide synthases in tendon overuse. J Orthop Res 24:80–86

  37. Tomiosso T, Nakagaki W, Gomes L, Hyslop S, Pimentel E (2009) Organization of collagen bundles during tendon healing in rats treated with L-NAME. Cell Tissue Res 337:235–242

  38. Twaddle BC, Poon P (2007) Early motion for Achilles tendon ruptures: is surgery important? Am J Sports Med 35:2033–2038

  39. Urtasun R, Cubero FJ, Vera M, Nieto N (2009) Reactive nitrogen species switch on early extracellular matrix remodeling via induction of MMP1 and TNFα. Gastroenterology 136:1410–1422

  40. Vallance P (2003) Nitric oxide: therapeutic opportunities. Fundam Clin Pharmacol 17:1–10

  41. Weber M, Niemann M, Lanz R, Müller T (2003) Nonoperative treatment of acute rupture of the Achilles tendon. Am J Sports Med 31:685–691

  42. Xia W, Wang Y, Appleyard R, Smythe G, Murrell G (2006) Spontaneous recovery of injured Achilles tendon in inducible nitric oxide synthase gene knockout mice. Inflamm Res 55:40–45

  43. Xu Y, Murrell G (2008) The basic science of tendinopathy. Clin Orthop Relat Res 466:1528–1538

Download references

Author information

Correspondence to Anderson M. Herculano.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Moraes, S.A.S., Oliveira, K.R.M., Crespo-López, M.E. et al. Local NO synthase inhibition produces histological and functional recovery in Achilles tendon of rats after tenotomy. Cell Tissue Res 353, 457–463 (2013). https://doi.org/10.1007/s00441-013-1662-2

Download citation

Keywords

  • Tendon injury
  • Nitric oxide
  • Tendon healing
  • Nitric oxide synthase
  • Wound healing