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Organization of collagen bundles during tendon healing in rats treated with L-NAME

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Abstract

The Achilles tendon can support high tension forces and may experience lesions. The damaged tissue does not regenerate completely, with the organization and mechanical properties of the repaired tendon being inferior to those of a healthy tendon. Nitric oxide (NO) plays an important role in wound repair. We have examined the structural reorganization and repair in Achilles tendon after injury in rats treated with the NO synthase inhibitor L-NAME. The right Achilles tendon of male Wistar rats was partially transected. One group of rats was treated with L-NAME (~300 mg/kg per day, given in drinking water) for 4 days prior to tendon sectioning and throughout the post-operative period. Control rats received water without L-NAME. The tendons were excised at 7, 14, and 21 days post-injury and used to quantify hydroxyproline and for mechanical tests. Tendons were also processed for histomorphological analysis by polarized light microscopy, which showed that the collagen fibers were disorganized by day 7 in non-treated and L-NAME-treated rats. In non-treated rats, the organization of the extracellular matrix was more homogeneous by days 14 and 21 compared with day 7, although this homogeneity was less than that in normal tendon. In contrast, in injured tendons from L-NAME-treated rats, the collagen fibers were still disorganized on day 21. Tendons from treated rats had more hydroxyproline but lower mechanical properties compared with those from non-treated rats. Thus, NO modulates tendon healing, with a reduction in NO biosynthesis delaying reorganization of the extracellular matrix, especially collagen.

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References

  • Albina JE, Mills CD, Henry WL, Caldwell MD (1990) Temporal expression of different pathways of L-arginine metabolism in healing wounds. J Immunol 144:3877–3880

    PubMed  CAS  Google Scholar 

  • Arti S, Anamika MR, Ravi S (1999) Nitric oxide inhibits wound collagen synthesis. Mol Cell Biochem 200:27–33

    Article  Google Scholar 

  • Bulgrin JP, Shabani M, Chakravarthy D, Smith DJ (1995) Nitric oxide synthesis is suppressed in steroid-impaired and diabetic wounds. Wounds 7:48–57

    Google Scholar 

  • Chakravortty D, Kumar KS (1997) Induction of cell proliferation and collagen synthesis in human small intestinal lamina propria fibroblasts by lipopolysaccharide: possible involvement of nitric oxide. Biochem Biophys Res Commun 240:458–463

    Article  PubMed  CAS  Google Scholar 

  • Chatziantoniou C, Boffa JJ, Ardoillou R, Dussaule JC (1998) Nitric oxide inhibition induces early activation of type I collagen gene in renal resistance vessels and glomeruli in transgenic mice. Role of endothelin. J Clin Invest 101:2780–2789

    Article  PubMed  CAS  Google Scholar 

  • Cribb M, Scott JE (1995) Tendon response to tensile stress: an ultrastructural investigation of collagen: proteoglycan interactions in stressed tendons. J Anat 187:423–428

    PubMed  Google Scholar 

  • Davis KL, Martin E, Turko IV, Murad F (2001) Novel effects of nitric oxide. Annu Rev Pharmacol Toxicol 41:203–236

    Article  PubMed  CAS  Google Scholar 

  • Ezura Y, Chakravarti S, Oldberg A, Chervoneva I, Birk DE (2000) Diferential expression of lumican and fibromodulin regulate collagen fibrilogenesis in developing mouse tendons. J Cell Biol 151:779–787

    Article  PubMed  CAS  Google Scholar 

  • Frank C, McDonald D, Wilson J, Eyre D, Shrive N (1995) Rabbit medial collateral ligament scar weakness is associated with decreased collagen pyridinoline crosslink density. J Orthop Res 13:157–165

    Article  PubMed  CAS  Google Scholar 

  • Griffith OW, Stuehr DJ (1995) Nitric oxide synthases: properties and catalytic mechanism. Annu Rev Physiol 57:707–736

    Article  PubMed  CAS  Google Scholar 

  • Hsu Y-C, Hsiao M, Wang L-F, Chien YW, Lee W-R (2006) Nitric oxide produced by iNOS is associated with collagen synthesis in keloid scar formation. Nitric Oxide 14:327–334

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Lin TW, Cardena L, Soslowsky LJ (2004) Biomechanics of tendon injury and repair. J Biomech 37:865–877

    Article  PubMed  Google Scholar 

  • Liu SH, Yang RS, al-Shaikh R, Lane JM (1995) Collagen in tendon, ligament, and bone healing. A current review. Clin Orthop Relat Res 318:265–278

    PubMed  Google Scholar 

  • Luo J, Chen AF (2005) Nitric oxide, a newly discovered function on wound healing. Acta Pharmacol Sin 26:256–264

    Article  Google Scholar 

  • Madden JW (1970) Current concepts of wound healing as applied to hand surgery. Orthop Clin North Am 1:325–334

    PubMed  CAS  Google Scholar 

  • Midwood KS, Williams LV, Schwarzbauer JE (2004) Tissue repair and the dynamics of the extracellular matrix. Int J Biochem Cell Biol 36:1031–1037

    Article  PubMed  CAS  Google Scholar 

  • Moncada S, Higgs EA (1993) The L-arginine-nitric oxide pathway. N Engl J Med 329:2001–2012

    Article  Google Scholar 

  • Murrell GAC (2007a) Using nitric oxide to treat tendinopathy. Br J Sports Med 41:227–231

    Article  PubMed  Google Scholar 

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

    Article  PubMed  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Muscara NM, McKnight W, Asfaha S, Wallace JL (2000) Wound collagen deposition in rats: effects of an NO-NSAID and a selective COX-2 inhibitor. Br J Pharmacol 129:681–686

    Article  PubMed  CAS  Google Scholar 

  • Nakagaki WR, Biancalana A, Benevides GP, Gomes L (2007) Biomechanical and biochemical properties of chicken calcaneal tendon under effect of age and nonforced active exercise. Connect Tissue Res 48:219–228

    Article  PubMed  CAS  Google Scholar 

  • Owens MW, Milligan SA, Grisham MB (1996) Inhibition of pleural mesothelial cell collagen synthesis by nitric oxide. Free Radic Biol Med 21:601–607

    Article  PubMed  CAS  Google Scholar 

  • Reddy GK, Stehno-Bittel L, Enwemeka CS (1999) Matrix remodeling in healing rabbit Achilles tendon. Wound Repair Regen 7:518–527

    Article  PubMed  CAS  Google Scholar 

  • Rizk M, Witte MB, Barbul A (2004) Nitric oxide and wound healing. World J Surg 28:301–306

    Article  PubMed  Google Scholar 

  • Schäffer MR, Tantry U, Gross SS, Wasserburg HL, Barbul A (1996) Nitric oxide regulates wound healing. J Surg Res 63:237–240

    Article  PubMed  Google Scholar 

  • Schäffer MR, Efron PA, Thornton FJ, Klingel K, Gross SS, Barbul A (1997) Nitric oxide, an autocrine regulator of wound fibroblast synthetic function. J Immunol 158:2375–2381

    PubMed  Google Scholar 

  • Schäffer MR, Tantry U, Thornton FJ, Barbul A (1999) Inhibition of nitric oxide synthesis in wounds: pharmacology and effect on accumulation of collagen in wounds in mice. Eur J Surg 165:262–267

    Article  PubMed  Google Scholar 

  • Sharma P, Maffulli N (2005) Tendon injury and tendinopathy: healing and repair. J Bone Joint Surg 87:187–202

    Article  PubMed  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  • Shi HP, Efron DT, Most D, Tantry US, Barbul A (2000) Supplemental dietary arginine enhances wound healing in normal but not inducible nitric oxide synthase knockout mice. Surgery 128:374–378

    Article  PubMed  CAS  Google Scholar 

  • Shi HP, Efron DT, Most D, Barbul A (2001) The role of iNOS in wound healing. Surgery 130:225–229

    Article  PubMed  CAS  Google Scholar 

  • Shukla A, Rasik AM, Shankar R (1999) Nitric oxide inhibits wound collagen synthesis. Mol Cell Biochem 200:27–33

    Article  PubMed  CAS  Google Scholar 

  • Silva MJ, Ritty TM, Ditsios K, Burns ME, Boyer MI, Gelberman RH (2007) Tendon injury response: assessment of biomechanical properties, tissue morphology and viability following flexor digitorum profundus tendon transection. J Orthop Res 22:990–997

    Article  Google Scholar 

  • Sparrow KJ, Finucane SD, Owen JR, Wayne JS (2005) The effects of low-intensity ultrasound on medial collateral ligament in the rabbit model. Am J Sports Med 33:1048–1056

    Article  PubMed  Google Scholar 

  • Stegeman H, Stalder K (1967) Determination of hydroxyproline. Clin Chim Acta 18:267–273

    Article  Google Scholar 

  • Thornton FJ, Schäffer MR, Witte MB, Moldawer LL, MacKay SL, Abouhamze A, Tannahill CL, Barbul A (1998) Enhanced collagen accumulation following direct transfection of the inducible nitric oxide synthase gene in cutaneous wounds. Biochem Biophys Res Commun 246:654–659

    Article  PubMed  CAS  Google Scholar 

  • Vidal BC (1995) Crimp as part of helical structure. C R Acad Sci III 318:173–178

    Google Scholar 

  • Viidik A (1978) On the correlation between structure and mechanical function of soft connective tissues. Verh Anat Ges 72:75–89

    PubMed  Google Scholar 

  • Vilarta R, Vidal BC (1989) Anisotropic and biomechanical properties of tendons modified by exercise and denervation: aggregation and macromolecular order in collagen bundles. Matrix 9:55–61

    PubMed  CAS  Google Scholar 

  • Witte MB, Barbul A (2002) Role of nitric oxide in wound repair. Am J Surg 4:406–412

    Article  Google Scholar 

  • Woo SL, Hildebrand K, Watanabe N, Fenwick JA, Papageorgiou CD, Wang JH (1999a) Tissue engineering of ligament and tendon healing. Clin Orthop Relat Res 367:S312–S323

    Article  PubMed  Google Scholar 

  • Woo SLY, Debski RE, Withrow JD, Janaushek MA (1999b) Biomechanics of knee ligaments. Am J Sports Med 27:533–543

    PubMed  CAS  Google Scholar 

  • Xia W, Szomor Z, Wang Y, Murrell GAC (2006a) Nitric oxide enhances collagen synthesis in cultured human tendon cells. J Orthop Res 24:159–172

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The authors thank Francisco Angelo Malatesta for technical assistance.

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Authors and Affiliations

  1. Department of Anatomy, Cell Biology, Physiology and Biophysics - Institute of Biology, State University of Campinas - UNICAMP, CP 6109, 13083–863, Campinas, SP, Brazil

    Tatiana Carla Tomiosso, Wilson Romero Nakagaki, Laurecir Gomes & Edson Rosa Pimentel

  2. Department of Pharmacology, School of Medical Sciences, State University of Campinas - UNICAMP, CP 6109, Campinas, 13083-863, SP, Brazil

    Stephen Hyslop

  3. Interdisciplinary Department - PURO, Fluminense Federal University - UFF, Rio das Ostras, RJ, Brazil

    Tatiana Carla Tomiosso

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  1. Tatiana Carla Tomiosso
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  2. Wilson Romero Nakagaki
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  3. Laurecir Gomes
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  4. Stephen Hyslop
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  5. Edson Rosa Pimentel
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Correspondence to Edson Rosa Pimentel.

Additional information

T.C.T. and W.R.N were supported by studentships from CAPES, and S.H. was supported by a research fellowship from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

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Tomiosso, T.C., Nakagaki, W.R., Gomes, L. et al. Organization of collagen bundles during tendon healing in rats treated with L-NAME. Cell Tissue Res 337, 235–242 (2009). https://doi.org/10.1007/s00441-009-0819-5

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  • DOI: https://doi.org/10.1007/s00441-009-0819-5

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