Advertisement

Calcified Tissue International

, Volume 82, Issue 4, pp 309–315 | Cite as

Trigeminal Nitric Oxide Synthase Expression Correlates with New Bone Formation During Distraction Osteogenesis

  • Rubens Ferreira de AlbuquerqueJr.
  • Elaine Aparecida Del Bel
  • Luiz Guilherme Brentegani
  • Maria Tereza Moura de Oliveira
  • João Paulo Mardegan Issa
Article

Abstract

Nitric oxide synthase (NOS) has been reported to be involved with both bone healing and bone metabolism. The aim of this study was to test the null hypothesis that there is no correlation between new bone formation during mandibular distraction osteogenesis and NOS expression in the trigeminal ganglion of rats. Newly formed tissue during distraction osteogenesis and trigeminal NOS expression measured by the NADPH-diaphorase (NADPH-d) reaction were evaluated in 72 male Wistar rats by histomorphometric and histochemical methods. In animals submitted to 0.5 mm/day distraction osteogenesis, the percentage of bone tissue was higher in the basal area of the mandibles compared with the center and significantly increased through the experimental periods (P < 0.05). At the sixth postoperative week, the difference in bone formation between the continuous and acute distraction osteogenesis groups was the highest. Significant correlation between new bone formation by distraction osteogenesis and NADPH-d-reactive neurons was found, varying according to neuronal cell size (r = −0.6, P = 0.005, small cells strongly stained; r = 0.5, P = 0.018, large cells moderately stained). The results suggest that NOS may play a role in the bone healing process via neurogenic pathways, and the phenomenon seems to be neuronal cell morphotype-dependent. Further studies are now warranted to investigate the mechanistic link between the expression of trigeminal NOS and mandibular new bone formation by distraction osteogenesis.

Keywords

Biomechanics Bone histology and histomorphometry Fracture repair Nitric oxide Orthopedics 

Notes

Acknowledgements

This study was supported by the Foundation for Research Support of São Paulo (FAPESP). We are grateful to Ms. Celia A. da Silva, Ms. Renata Ferreira da Silva and Dr. José Ferreira de Carvalho for their generous technical assistance.

References

  1. 1.
    Ilizarov GA (1990) Clinical application of the tension-stress effect for limb lengthening. Clin Orthop Relat Res 250:8–26PubMedGoogle Scholar
  2. 2.
    Faber J, Azevedo RB, Bao SN (2005) Distraction osteogenesis may promote periodontal bone regeneration. J Dent Res 84:757–761PubMedGoogle Scholar
  3. 3.
    Rowe NM, Mehrara BJ, Luchs JS, Dudziak ME, Steinbrech DS, Illei PB, Fernandez GJ, Gittes GK, Longaker MT (1999) Angiogenesis during mandibular distraction osteogenesis. Ann Plast Surg 42:470–475PubMedCrossRefGoogle Scholar
  4. 4.
    Wang LC, Takahashi I, Sasano Y, Sugawara J, Mitani H (2005) Osteoclastogenic activity during mandibular distraction osteogenesis. J Dent Res 84:1010–1015PubMedGoogle Scholar
  5. 5.
    Liu ZJ, King GJ, Herring SW (2006) Condylar mineralization following mandibular distraction in rats. J Dent Res 85:653–657PubMedCrossRefGoogle Scholar
  6. 6.
    van’t Hof RJ, Ralston SH (2001) Nitric oxide and bone. Immunology 103:255–261PubMedCrossRefGoogle Scholar
  7. 7.
    Nakahara S, Yone K, Setoguchi T, Yamaura I, Arishima Y, Yoshino S, Komiya S (2002) Changes in nitric oxide and expression of nitric oxide synthase in spinal cord after acute traumatic injury in rats. J Neurotrauma 19:1467–1474PubMedCrossRefGoogle Scholar
  8. 8.
    van’t Hof RJ, Macphee J, Libouban H, Helfrich MH, Ralston SH (2004) Regulation of bone mass and bone turnover by neuronal nitric oxide synthase. Endocrinology 145:5068–5074PubMedCrossRefGoogle Scholar
  9. 9.
    Huitema LF, Vaandrager AB, Van Weeren PR, Barneveld A, Helms JB, Van De Lest CH (2006) The nitric oxide donor sodium nitroprusside inhibits mineralization in ATDC5 cells. Calcif Tissue Int 78:171–177PubMedCrossRefGoogle Scholar
  10. 10.
    Snyder SH, Bredt DS (1991) Nitric oxide as a neuronal messenger. Trends Pharmacol Sci 12:125–128PubMedCrossRefGoogle Scholar
  11. 11.
    Moncada S, Higgs A (1993) The l-arginine-nitric oxide pathway. N Engl J Med 329:2002–2012PubMedCrossRefGoogle Scholar
  12. 12.
    Dawson TM, Bredt DS, Fotuhi M, Hwang PM, Snyder SH (1991) Nitric oxide synthase and neuronal NADPH diaphorase are identical in brain and peripheral tissues. Proc Natl Acad Sci USA 88:7797–7801PubMedCrossRefGoogle Scholar
  13. 13.
    Hope BT, Michael GJ, Knigge KM, Vincent SR (1991) Neuronal NADPH-diaphorase is a nitric oxide synthase. Proc Natl Acad Sci USA 88:2811–2814PubMedCrossRefGoogle Scholar
  14. 14.
    Nomura S, Takano-Yamamoto T (2000) Molecular events caused by mechanical stress in bone. Matrix Biol 19:91–96PubMedCrossRefGoogle Scholar
  15. 15.
    Baldik Y, Talu U, Altinel L, Bilge H, Demiryont M, Aykac-Toker G (2002) Bone healing regulated by nitric oxide: an experimental study in rats. Clin Orthop Relat Res 404:343–352PubMedCrossRefGoogle Scholar
  16. 16.
    Baldik Y, Diwan AD, Appleyard RC, Fang ZM, Wang Y, Murrell GA (2005) Deletion of iNOS gene impairs mouse fracture healing. Bone 37:32–36PubMedCrossRefGoogle Scholar
  17. 17.
    Johnson DL, Mcallister TN, Frangos JA (1996) Fluid flow stimulates rapid and continuous release of nitric oxide in osteoblasts. Am J Physiol Endocrinol Metab 271:E205–E208Google Scholar
  18. 18.
    Klein-Nulend J, Helfrich MH, Sterck JG, Macpherson H, Joldersma M, Ralston SH, Semeins CM, Burger EH (1998) Nitric oxide response to shear stress by human bone cell cultures is endothelial nitric oxide synthase dependent. Biochem Biophys Res Commun 250:108–114PubMedCrossRefGoogle Scholar
  19. 19.
    Burger EH, Klein-Nulen J (1999) Responses of bone cells to biomechanical forces in vitro. Adv Dent Res 13:93–98PubMedCrossRefGoogle Scholar
  20. 20.
    Davies SL, Loescher AR, Clayton NM, Bountra C, Robinson PP, Boissonade FM (2004) nNOS expression following inferior alveolar nerve injury in the ferret. Brain Res 1027:11–17PubMedCrossRefGoogle Scholar
  21. 21.
    Banks RW (1999) Histological staining methods. In: Johansson H, Windhorst U (eds) Modern techniques in neuroscience research. Springer-Verlag, New York, pp 437–458Google Scholar
  22. 22.
    Echeverry MB, Guimaraes FS, Del Bel EA (2004) Acute and delayed restraint stress-induced changes in nitric oxide producing neurons in limbic regions. Neuroscience 125:981–993PubMedCrossRefGoogle Scholar
  23. 23.
    Paxinos G, Watson C (1998) The rat brain in stereotaxic coordinates. Academic Press, San DiegoGoogle Scholar
  24. 24.
    Weibel ER, Kistler GS, Scherle WF (1966) Practical stereological methods for morphometric cytology. J Cell Biol 30:23–38PubMedCrossRefGoogle Scholar
  25. 25.
    Fisher RA (1925) Statistical methods for research workers. Oliver and Boyd, EdinburghGoogle Scholar
  26. 26.
    Lee PC, Salyapongse AN, Bragdon GA, Shears LL, Watkins SC, Edington HD, Billiar TR (1999) Impaired wound healing and angiogenesis in eNOS-deficient mice. Am J Physiol Heart Circ Physiol 277:H1600–H1608Google Scholar
  27. 27.
    Zhang X, Ji RR, Arvidsson J, Lundberg JM, Bartfai T, Bedecs K, Hökfelt T (1996) Expression of peptides, nitric oxide synthase and NPY receptor in trigeminal and nodose ganglia after nerve lesions. Exp Brain Res 111:393–404PubMedCrossRefGoogle Scholar
  28. 28.
    Levy D, Zochodne DW (1998) Local nitric oxide synthase activity in a model of neuropathic pain. Eur J Neurosci 10:1846–1855PubMedCrossRefGoogle Scholar
  29. 29.
    Luo ZD, Chaplan SR, Scott BP, Cizkova D, Calcutt NA, Yaksh TL (1999) Neuronal nitric oxide synthase mRNA upregulation in rat sensory neurons after spinal nerve ligation: lack of a role in allodynia development. J Neurosci 19:9201–9208PubMedGoogle Scholar
  30. 30.
    Thippeswamy T, Jain RK, Mumtaz N, Morris R (2001) Inhibition of neuronal nitric oxide synthase results in neurodegenerative changes in the axotomised dorsal root ganglion neurons: evidence for a neuroprotective role of nitric oxide in vivo. Neurosci Res 40:37–44PubMedCrossRefGoogle Scholar
  31. 31.
    Yonehara N, Takemura M, Shigenaga Y (1997) Involvement of nitric oxide in re-innervation of rat molar tooth pulp following transection of the inferior alveolar nerve. Brain Res 757:31–36PubMedCrossRefGoogle Scholar
  32. 32.
    Koltzenburg M, Wall PD, Mcmahon SB (1999) Does the right side know what the left is doing? Trends Neurosci 22:122–127PubMedCrossRefGoogle Scholar
  33. 33.
    Tao YX, Johns RA (2002) Activation and up-regulation of spinal cord nitric oxide receptor, soluble guanylate cyclase, after formalin injection into the rat hind paw. Neuroscience 112:439–446PubMedCrossRefGoogle Scholar
  34. 34.
    Pose I, Sampogna S, Chase MH, Morales FR (2003) Mesencephalic trigeminal neurons are innervated by nitric oxide synthase-containing fibers and respond to nitric oxide. Brain Res 960:81–89PubMedCrossRefGoogle Scholar
  35. 35.
    Tassorelli C, Greco R, Wang D, Sandrini G, Nappi G (2006) Prostaglandins, glutamate and nitric oxide synthase mediate nitroglycerin-induced hyperalgesia in the formalin test. Eur J Pharmacol 534:103–107PubMedCrossRefGoogle Scholar
  36. 36.
    Fong AY, Talman WT, Lawrence AJ (2000) Axonal transport of NADPH-diaphorase and [3H]nitro-l-arginine binding, but not [(3)H]cGMP binding, by the rat vagus nerve. Brain Res 878:240–246PubMedCrossRefGoogle Scholar
  37. 37.
    Gao S, Fei M, Cheng C, Yu X, Chen M, Shi S, Qin J, Guo Z, Shen A (2007) Spatiotemporal expression of PSD-95 and nNOS after rat sciatic nerve injury. Neurochem Res. doi:  10.1007/s11064-007-9555-y. Cited 20 Dec, 2007
  38. 38.
    Issa JPM, Nascimento C, Bentley MVLB, Guimarães EADBB, Iyomasa MM, Sebald W, Albuquerque RF Jr (2007) Bone repair in rat mandible by rhBMP-2 associated with two carriers. Micron (in press). doi:  10.1016/j.micron.2007.03.008. Cited 24 March, 2007

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Rubens Ferreira de AlbuquerqueJr.
    • 1
  • Elaine Aparecida Del Bel
    • 1
  • Luiz Guilherme Brentegani
    • 1
  • Maria Tereza Moura de Oliveira
    • 1
  • João Paulo Mardegan Issa
    • 1
  1. 1.Faculty of Dentistry of Ribeirão PretoUniversity of São PauloSão PauloBrazil

Personalised recommendations