European Spine Journal

, Volume 27, Issue 4, pp 739–751 | Cite as

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

  • 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 MasudaEmail author
Original Article



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.


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.


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.


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

Graphical Abstract


Intervertebral disc degeneration Growth differentiation factor-6 Rabbit anular-puncture model Rat xenograft radiculopathy model Pain behavior Inflammatory cytokines 



The authors would like to thank Erikka Linn, Edward Abarado, and Weibo Jiang, M.D. for their generous assistance.

Compliance with ethical standards

Disclosures/Conflicts of interest

The authors did not receive outside funding or support for this work, and have no actual or potential conflicts of interest to disclose.

Supplementary material

586_2018_5488_MOESM1_ESM.pptx (343 kb)
Supplementary material 1 (PPTX 342 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Shingo Miyazaki
    • 1
    • 2
  • Ashish D. Diwan
    • 3
  • Kenji Kato
    • 1
    • 4
  • Kevin Cheng
    • 1
  • Won C. Bae
    • 5
  • Yang Sun
    • 6
    • 7
  • Junichi Yamada
    • 1
    • 8
  • Carol Muehleman
    • 9
  • Mary E. Lenz
    • 1
  • Nozomu Inoue
    • 10
  • Robert L. Sah
    • 1
    • 6
  • Mamoru Kawakami
    • 11
  • Koichi Masuda
    • 1
    Email author
  1. 1.Department of Orthopaedic SurgeryUniversity of California-San DiegoLa JollaUSA
  2. 2.Department of Orthopaedic SurgeryKobe University Graduate School of MedicineKobeJapan
  3. 3.Orthopaedic Research Institute and Department of Orthopaedic SurgerySt George Hospital, University of New South WalesSydneyAustralia
  4. 4.Department of Orthopaedic SurgeryNagoya City University Graduate School of MedicineNagoyaJapan
  5. 5.Department of RadiologyUniversity of California-San DiegoLa JollaUSA
  6. 6.Department of BioengineeringUniversity of California-San DiegoLa JollaUSA
  7. 7.Orthopaedic Medical Centerthe Second Hospital of Jilin UniversityChangchunChina
  8. 8.Department of Orthopaedic SurgeryMie University Graduate School of MedicineTsuJapan
  9. 9.Department of BiochemistryRush University Medical CenterChicagoUSA
  10. 10.Department of Orthopaedic SurgeryRush University Medical CenterChicagoUSA
  11. 11.Spine Care CenterWakayama Medical University Kihoku HospitalKatsuragicho, Ito GunJapan

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