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01-04-2018 | Original Article

ISSLS PRIZE IN BASIC SCIENCE 2018: Growth differentiation factor-6 attenuated pro-inflammatory molecular changes in the rabbit anular-puncture model and degenerated disc-induced pain generation in the rat xenograft radiculopathy model

Authors: Shingo Miyazaki, Ashish D. Diwan, Kenji Kato, Kevin Cheng, Won C. Bae, Yang Sun, Junichi Yamada, Carol Muehleman, Mary E. Lenz, Nozomu Inoue, Robert L. Sah, Mamoru Kawakami, Koichi Masuda

Published in: European Spine Journal | Issue 4/2018

Abstract

Purpose

To elucidate the effects of growth differentiation factor-6 (GDF6) on: (i) gene expression of inflammatory/pain-related molecules and structural integrity in the rabbit intervertebral disc (IVD) degeneration model, and (ii) sensory dysfunction and changes in pain-marker expression in dorsal nerve ganglia (DRGs) in the rat xenograft radiculopathy model.

Methods

Forty-six adolescent rabbits received anular-puncture in two non-consecutive lumbar IVDs. Four weeks later, phosphate-buffered saline (PBS) or GDF6 (1, 10 or 100 µg) was injected into the nucleus pulposus (NP) of punctured discs and followed for 4 weeks for gene expression analysis and 12 weeks for structural analyses. For pain assessment, eight rabbits were sacrificed at 4 weeks post-injection and NP tissues of injected discs were transplanted onto L5 DRGs of 16 nude rats to examine mechanical allodynia. The rat DRGs were analyzed immunohistochemically.

Results

In GDF6-treated rabbit NPs, gene expressions of interleukin-6, tumor necrosis factor-α, vascular endothelial growth factor, prostaglandin-endoperoxide synthase 2, and nerve growth factor were significantly lower than those in the PBS group. GDF6 injections resulted in partial restoration of disc height and improvement of MRI disc degeneration grades with statistical significance in rabbit structural analyses. Allodynia induced by xenograft transplantation of rabbit degenerated NPs onto rat DRGs was significantly reduced by GDF6 injection. Staining intensities for ionized calcium-binding adaptor molecule-1 and calcitonin gene-related peptide in rat DRGs of the GDF6 group were significantly lower than those of the PBS group.

Conclusion

GDF6 injection may change the pathological status of degenerative discs and attenuate degenerated IVD-induced pain.

Graphical Abstract

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Andersson GB (1999) Epidemiological features of chronic low-back pain. Lancet 354:581–585CrossRefPubMed

Gilbertson L, Ahn SH, Teng PN, Studer RK, Niyibizi C, Kang JD (2008) The effects of recombinant human bone morphogenetic protein-2, recombinant human bone morphogenetic protein-12, and adenoviral bone morphogenetic protein-12 on matrix synthesis in human annulus fibrosis and nucleus pulposus cells. Spine J 8:449–456. https://doi.org/10.1016/j.spinee.2006.11.006 CrossRefPubMed

Le Maitre CL, Freemont AJ, Hoyland JA (2004) Localization of degradative enzymes and their inhibitors in the degenerate human intervertebral disc. J Pathol 204:47–54CrossRefPubMed

Abe Y, Akeda K, An HS, Aoki Y, Pichika R, Muehleman C, Kimura T, Masuda K (2007) Proinflammatory cytokines stimulate the expression of nerve growth factor by human intervertebral disc cells. Spine (Phila Pa 1976) 32:635–642. https://doi.org/10.1097/01.brs.0000257556.90850.53 CrossRef

Binch AL, Cole AA, Breakwell LM, Michael AL, Chiverton N, Cross AK, Le Maitre CL (2014) Expression and regulation of neurotrophic and angiogenic factors during human intervertebral disc degeneration. Arthritis Res Ther 16:416. https://doi.org/10.1186/s13075-014-0416-1 CrossRefPubMed

Lee JM, Song JY, Baek M, Jung HY, Kang H, Han IB, Kwon YD, Shin DE (2011) Interleukin-1beta induces angiogenesis and innervation in human intervertebral disc degeneration. J Orthop Res 29:265–269. https://doi.org/10.1002/jor.21210 CrossRefPubMed

Freemont AJ, Watkins A, Le Maitre C, Baird P, Jeziorska M, Knight MT, Ross ER, O’brien JP, Hoyland JA (2002) Nerve growth factor expression and innervation of the painful intervertebral disc. J Pathol 197:286–292. https://doi.org/10.1002/path.1108 CrossRefPubMed

Krock E, Rosenzweig DH, Chabot-Dore AJ, Jarzem P, Weber MH, Ouellet JA, Stone LS, Haglund L (2014) Painful, degenerating intervertebral discs up-regulate neurite sprouting and CGRP through nociceptive factors. J Cell Mol Med 18:1213–1225. https://doi.org/10.1111/jcmm.12268 CrossRefPubMedPubMedCentral

Purmessur D, Freemont AJ, Hoyland JA (2008) Expression and regulation of neurotrophins in the nondegenerate and degenerate human intervertebral disc. Arthritis Res Ther 10:R99. https://doi.org/10.1186/ar2487 CrossRefPubMedPubMedCentral

10.

Aoki Y, An HS, Takahashi K, Miyamoto K, Lenz ME, Moriya H, Masuda K (2007) Axonal growth potential of lumbar dorsal root ganglion neurons in an organ culture system: response of nerve growth factor-sensitive neurons to neuronal injury and an inflammatory cytokine. Spine (Phila Pa 1976) 32:857–863. https://doi.org/10.1097/01.brs.0000259810.48681.90 CrossRef

11.

Aoki Y, Takahashi Y, Ohtori S, Moriya H, Takahashi K (2004) Distribution and immunocytochemical characterization of dorsal root ganglion neurons innervating the lumbar intervertebral disc in rats: a review. Life Sci 74:2627–2642CrossRefPubMed

12.

Masuda K, Imai Y, Okuma M, Muehleman C, Nakagawa K, Akeda K, Thonar E, Andersson G, An HS (2006) Osteogenic protein-1 injection into a degenerated disc induces the restoration of disc height and structural changes in the rabbit anular puncture model. Spine (Phila Pa 1976) 31:742–754. https://doi.org/10.1097/01.brs.0000206358.66412.7b CrossRef

13.

Chujo T, An HS, Akeda K, Miyamoto K, Muehleman C, Attawia M, Andersson G, Masuda K (2006) Effects of growth differentiation factor-5 on the intervertebral disc–in vitro bovine study and in vivo rabbit disc degeneration model study. Spine (Phila Pa 1976) 31:2909–2917. https://doi.org/10.1097/01.brs.0000248428.22823.86 CrossRef

14.

Masuda K, Oegema TR Jr, An HS (2004) Growth factors and treatment of intervertebral disc degeneration. Spine (Phila Pa 1976) 29:2757–2769CrossRef

15.

Chang SC, Hoang B, Thomas JT, Vukicevic S, Luyten FP, Ryba NJ, Kozak CA, Reddi AH, Moos M Jr (1994) Cartilage-derived morphogenetic proteins. New members of the transforming growth factor-beta superfamily predominantly expressed in long bones during human embryonic development. J Biol Chem 269:28227–28234PubMed

16.

Shen B, Bhargav D, Wei A, Williams LA, Tao H, Ma DD, Diwan AD (2009) BMP-13 emerges as a potential inhibitor of bone formation. Int J Biol Sci 5:192–200CrossRefPubMedPubMedCentral

17.

Wei A, Shen B, Williams LA, Bhargav D, Gulati T, Fang Z, Pathmanandavel S, Diwan AD (2016) Expression of growth differentiation factor 6 in the human developing fetal spine retreats from vertebral ossifying regions and is restricted to cartilaginous tissues. J Orthop Res 34:279–289. https://doi.org/10.1002/jor.22983 CrossRefPubMed

18.

Williams LA, Bhargav D, Diwan AD (2008) Unveiling the bmp13 enigma: redundant morphogen or crucial regulator? Int J Biol Sci 4:318–329CrossRefPubMedPubMedCentral

19.

Gulati T, Chung SA, Wei AQ, Diwan AD (2015) Localization of bone morphogenetic protein 13 in human intervertebral disc and its molecular and functional effects in vitro in 3D culture. J Orthop Res 33:1769–1775. https://doi.org/10.1002/jor.22965 CrossRefPubMed

20.

Wei A, Williams LA, Bhargav D, Shen B, Kishen T, Duffy N, Diwan AD (2009) BMP13 prevents the effects of annular injury in an ovine model. Int J Biol Sci 5:388–396CrossRefPubMedPubMedCentral

21.

Kawakami M, Tamaki T, Hayashi N, Hashizume H, Matsumoto T, Minamide A, Kihira T (2000) Mechanical compression of the lumbar nerve root alters pain-related behaviors induced by the nucleus pulposus in the rat. J Orthop Res 18:257–264. https://doi.org/10.1002/jor.1100180214 CrossRefPubMed

22.

Masuda K, Aota Y, Muehleman C, Imai Y, Okuma M, Thonar EJ, Andersson GB, An HS (2005) A novel rabbit model of mild, reproducible disc degeneration by an anulus needle puncture: correlation between the degree of disc injury and radiological and histological appearances of disc degeneration. Spine (Phila Pa 1976) 30:5–14CrossRef

23.

Mwale F, Masuda K, Pichika R, Epure LM, Yoshikawa T, Hemmad A, Roughley PJ, Antoniou J (2011) The efficacy of Link N as a mediator of repair in a rabbit model of intervertebral disc degeneration. Arthritis Res Ther 13:R120. https://doi.org/10.1186/ar3423 CrossRefPubMedPubMedCentral

24.

Espinoza Orias AA, Mammoser NM, Triano JJ, An HS, Andersson GB, Inoue N (2016) Effects of axial torsion on disc height distribution: an in vivo study. J Manip Physiol Ther 39:294–303. https://doi.org/10.1016/j.jmpt.2016.03.002 CrossRef

25.

Pfirrmann CW, Metzdorf A, Zanetti M, Hodler J, Boos N (2001) Magnetic resonance classification of lumbar intervertebral disc degeneration. Spine (Phila Pa 1976) 26:1873–1878CrossRef

26.

Dixon WJ (1980) Efficient analysis of experimental observations. Annu Rev Pharmacol Toxicol 20:441–462. https://doi.org/10.1146/annurev.pa.20.040180.002301 CrossRefPubMed

27.

Chaplan SR, Bach FW, Pogrel JW, Chung JM, Yaksh TL (1994) Quantitative assessment of tactile allodynia in the rat paw. J Neurosci Methods 53:55–63. https://doi.org/10.1016/0165-0270(94)90144-9 CrossRefPubMed

28.

Le Maitre CL, Freemont AJ, Hoyland JA (2005) The role of interleukin-1 in the pathogenesis of human intervertebral disc degeneration. Arthritis Res Ther 7:R732–R745CrossRefPubMedPubMedCentral

29.

Weiler C, Nerlich AG, Bachmeier BE, Boos N (2005) Expression and distribution of tumor necrosis factor alpha in human lumbar intervertebral discs: a study in surgical specimen and autopsy controls. Spine (Phila Pa 1976) 30:44–53 (discussion 54) CrossRef

30.

Clarke LE, McConnell JC, Sherratt MJ, Derby B, Richardson SM, Hoyland JA (2014) Growth differentiation factor 6 and transforming growth factor-beta differentially mediate mesenchymal stem cell differentiation, composition, and micromechanical properties of nucleus pulposus constructs. Arthritis Res Ther 16:R67. https://doi.org/10.1186/ar4505 CrossRefPubMedPubMedCentral

Title: ISSLS PRIZE IN BASIC SCIENCE 2018: Growth differentiation factor-6 attenuated pro-inflammatory molecular changes in the rabbit anular-puncture model and degenerated disc-induced pain generation in the rat xenograft radiculopathy model
Authors: Shingo Miyazaki
Ashish D. Diwan
Kenji Kato
Kevin Cheng
Won C. Bae
Yang Sun
Junichi Yamada
Carol Muehleman
Mary E. Lenz
Nozomu Inoue
Robert L. Sah
Mamoru Kawakami
Koichi Masuda
Publication date: 01-04-2018
Publisher: Springer Berlin Heidelberg
Published in: European Spine Journal / Issue 4/2018
Print ISSN: 0940-6719
Electronic ISSN: 1432-0932
DOI: https://doi.org/10.1007/s00586-018-5488-1

Keynote webinar | Spotlight on medication adherence

Springer Medicine

ISSLS PRIZE IN BASIC SCIENCE 2018: Growth differentiation factor-6 attenuated pro-inflammatory molecular changes in the rabbit anular-puncture model and degenerated disc-induced pain generation in the rat xenograft radiculopathy model

Abstract

Purpose

Methods

Results

Conclusion

Graphical Abstract

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Graphical Abstract

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