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Intervertebral disc degenerative changes after intradiscal injection of TNF-α in a porcine model

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Abstract

Purpose

To investigate whether exogenous tumor necrosis factor-α (TNF-α) will initiate a degenerative process in intervertebral disc in vivo.

Methods

Exogenous TNF-α in dosages of 50 and 100 ng in 50 μL Dulbecco’s Modified Essential Medium (DMEM) was injected into porcine lumbar discs; a third disc was injected only with 50 μL DMEM as a control. Magnetic resonance imaging (MRI) yielding T1- and T2-weighted images, T2-mapping, and post-contrast T1 images was performed and histology was studied as well.

Results

After 3 months, a significant decrease in T2 value calculated from T2-mapping MRI was observed in the annulus and nucleus of both groups injected with TNF-α along with a slight decrease in disc height and nucleus volumes in comparison to the control discs. No obvious visual differences among the groups were observed in the normal T1- and T2-weighted MRI images. Post-contrast T1 MRI showed increased annulus enhancement in both TNF-α-injected groups compared to the control discs, while no enhancement difference was observed in the nucleus. Histological analysis showed degenerative changes with annulus fissure, cell cluster, nucleus matrix loss, vascularization and interleukin-1β expression in the outer annulus of both TNF-α-injected discs, while no degenerative changes were observed in the control discs.

Conclusions

Intradiscal injection of exogenous TNF-α caused early stage disc degeneration in a porcine model. It may thus support the hypothesis of exogenic TNF-α being an important early pathogenetic factor in disc degeneration.

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References

  1. Ohtori S, Inoue G, Eguchi Y, Orita S, Takaso M, Ochiai N, Kishida S, Kuniyoshi K, Aoki Y, Nakamura J, Ishikawa T, Arai G, Miyagi M, Kamoda H, Suzuki M, Sakuma Y, Oikawa Y, Kubota G, Inage K, Sainoh T, Toyone T, Yamauchi K, Kotani T, Akazawa T, Minami S, Takahashi K (2013) Tumor necrosis factor-alpha-immunoreactive cells in nucleus pulposus in adolescent patients with lumbar disc herniation. Spine 38:459–462. doi:10.1097/BRS.0b013e3182739cb4

    Article  PubMed  Google Scholar 

  2. Kato T, Haro H, Komori H, Shinomiya K (2004) Sequential dynamics of inflammatory cytokine, angiogenesis inducing factor and matrix degrading enzymes during spontaneous resorption of the herniated disc. J Orthop Res 22:895–900. doi:10.1016/j.orthres.2003.11.008

    Article  CAS  PubMed  Google Scholar 

  3. Hayashi S, Taira A, Inoue G, Koshi T, Ito T, Yamashita M, Yamauchi K, Suzuki M, Takahashi K, Ohtori S (2008) TNF-alpha in nucleus pulposus induces sensory nerve growth: a study of the mechanism of discogenic low back pain using TNF-alpha-deficient mice. Spine 33:1542–1546. doi:10.1097/BRS.0b013e318178e5ea

    Article  PubMed  Google Scholar 

  4. Risbud MV, Shapiro IM (2014) Role of cytokines in intervertebral disc degeneration: pain and disc content. Nat Rev Rheumatol 10:44–56. doi:10.1038/nrrheum.2013.160

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Markova DZ, Kepler CK, Addya S, Murray HB, Vaccaro AR, Shapiro IM, Anderson DG, Albert TJ, Risbud MV (2013) An organ culture system to model early degenerative changes of the intervertebral disc II: profiling global gene expression changes. Arthritis Res Ther 15:R121. doi:10.1186/ar4301

    Article  PubMed Central  PubMed  Google Scholar 

  6. Kim HJ, Yeom JS, Koh YG, Yeo JE, Kang KT, Kang YM, Chang BS, Lee CK (2014) Anti-inflammatory effect of platelet-rich plasma on nucleus pulposus cells with response of TNF-alpha and IL-1. J Orthop Res 32:551–556. doi:10.1002/jor.22532

    Article  CAS  PubMed  Google Scholar 

  7. Purmessur D, Walter BA, Roughley PJ, Laudier DM, Hecht AC, Iatridis J (2013) A role for TNFalpha in intervertebral disc degeneration: a non-recoverable catabolic shift. Biochem Biophys Res Commun 433:151–156. doi:10.1016/j.bbrc.2013.02.034

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  8. Ponnappan RK, Markova DZ, Antonio PJ, Murray HB, Vaccaro AR, Shapiro IM, Anderson DG, Albert TJ, Risbud MV (2011) An organ culture system to model early degenerative changes of the intervertebral disc. Arthritis Res Ther 13:R171. doi:10.1186/ar3494

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Chen S, Huang Y, Zhou ZJ, Hu ZJ, Wang JY, Xu WB, Fang XQ, Fan SW (2014) Upregulation of TNF-alpha and ADAMTS-5, but not ADAMTS-4, in human intervertebral cartilage endplate with Modic changes. Spine. doi:10.1097/BRS.0000000000000362

    Google Scholar 

  10. Luoma K, Vehmas T, Gronblad M, Kerttula L, Kaapa E (2009) Relationship of Modic type 1 change with disc degeneration: a prospective MRI study. Skeletal Radiol 38:237–244. doi:10.1007/s00256-008-0611-8

    Article  PubMed  Google Scholar 

  11. Kuisma M, Karppinen J, Niinimaki J, Kurunlahti M, Haapea M, Vanharanta H, Tervonen O (2006) A three-year follow-up of lumbar spine endplate (Modic) changes. Spine 31:1714–1718. doi:10.1097/01.brs.0000224167.18483.14

    Article  PubMed  Google Scholar 

  12. Kinra P, Dutta V (2013) Serum TNF alpha levels: a prognostic marker for assessment of severity of malaria. Trop Biomed 30:645–653

    CAS  PubMed  Google Scholar 

  13. Zygner W, Gojska-Zygner O, Baska P, Dlugosz E (2014) Increased concentration of serum TNF alpha and its correlations with arterial blood pressure and indices of renal damage in dogs infected with Babesia canis. Parasitol Res 113:1499–1503. doi:10.1007/s00436-014-3792-1

    Article  PubMed Central  PubMed  Google Scholar 

  14. Bruunsgaard H, Andersen-Ranberg K, Jeune B, Pedersen AN, Skinhoj P, Pedersen BK (1999) A high plasma concentration of TNF-alpha is associated with dementia in centenarians. J Gerontol Ser A Biol Sci Med Sci 54:M357–M364

    Article  CAS  Google Scholar 

  15. Bendtsen M, Bunger CE, Zou X, Foldager C, Jorgensen HS (2011) Autologous stem cell therapy maintains vertebral blood flow and contrast diffusion through the endplate in experimental intervertebral disc degeneration. Spine 36:E373–E379. doi:10.1097/BRS.0b013e3181dce34c

    Article  PubMed  Google Scholar 

  16. Obata S, Akeda K, Imanishi T, Masuda K, Bae W, Morimoto R, Asanuma Y, Kasai Y, Uchida A, Sudo A (2012) Effect of autologous platelet-rich plasma-releasate on intervertebral disc degeneration in the rabbit anular puncture model: a preclinical study. Arthritis Res Ther 14:R241. doi:10.1186/ar4084

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Imai Y, Okuma M, An HS, Nakagawa K, Yamada M, Muehleman C, Thonar E, Masuda K (2007) Restoration of disc height loss by recombinant human osteogenic protein-1 injection into intervertebral discs undergoing degeneration induced by an intradiscal injection of chondroitinase ABC. Spine 32:1197–1205. doi:10.1097/Brs.0b013e3180574d26

    Article  PubMed  Google Scholar 

  18. Rajasekaran S, Babu JN, Arun R, Armstrong BR, Shetty AP, Murugan S (2004) ISSLS prize winner: a study of diffusion in human lumbar discs: a serial magnetic resonance imaging study documenting the influence of the endplate on diffusion in normal and degenerate discs. Spine 29:2654–2667

    Article  CAS  PubMed  Google Scholar 

  19. Sive JI, Baird P, Jeziorsk M, Watkins A, Hoyland JA, Freemont AJ (2002) Expression of chondrocyte markers by cells of normal and degenerate intervertebral discs. Mol Pathol 55:91–97

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  20. Hauge EM, Qvesel D, Eriksen EF, Mosekilde L, Melsen F (2001) Cancellous bone remodeling occurs in specialized compartments lined by cells expressing osteoblastic markers. J Bone Miner Res 16:1575–1582. doi:10.1359/jbmr.2001.16.9.1575

    Article  CAS  PubMed  Google Scholar 

  21. Elliott DM, Yerramalli CS, Beckstein JC, Boxberger JI, Johannessen W, Vresilovic EJ (2008) The effect of relative needle diameter in puncture and sham injection animal models of degeneration. Spine 33:588–596. doi:10.1097/BRS.0b013e318166e0a2

    Article  PubMed  Google Scholar 

  22. Wang JL, Tsai YC, Wang YH (2007) The leakage pathway and effect of needle gauge on degree of disc injury post anular puncture: a comparative study using aged human and adolescent porcine discs. Spine 32:1809–1815. doi:10.1097/BRS.0b013e31811ec282

    Article  PubMed  Google Scholar 

  23. Jim B, Steffen T, Moir J, Roughley P, Haglund L (2011) Development of an intact intervertebral disc organ culture system in which degeneration can be induced as a prelude to studying repair potential. Eur Spine J 20:1244–1254. doi:10.1007/s00586-011-1721-x

    Article  PubMed Central  PubMed  Google Scholar 

  24. Mwale F, Demers CN, Michalek AJ, Beaudoin G, Goswami T, Beckman L, Iatridis JC, Antoniou J (2008) Evaluation of quantitative magnetic resonance imaging, biochemical and mechanical properties of trypsin-treated intervertebral discs under physiological compression loading. J Magn Reson Imaging 27:563–573. doi:10.1002/jmri.21242

    Article  PubMed Central  PubMed  Google Scholar 

  25. Pfirrmann CWA, Metzdorf A, Zanetti M, Hodler J, Boos N (2001) Magnetic resonance classification of lumbar intervertebral disc degeneration. Spine 26:1873–1878. doi:10.1097/00007632-200109010-00011

    Article  CAS  PubMed  Google Scholar 

  26. Rajasekaran S, Venkatadass K, Babu JN, Ganesh K, Shetty AP (2008) Pharmacological enhancement of disc diffusion and differentiation of healthy, ageing and degenerated discs. Eur Spine J 17:626–643. doi:10.1007/S00586-008-0645-6

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Gries NC, Berlemann U, Moore RJ, Vernon-Roberts B (2000) Early histologic changes in lower lumbar discs and facet joints and their correlation. Eur Spine J 9:23–29. doi:10.1007/S005860050004

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  28. Gonzalez-Quintela A, Campos J, Loidi L, Quinteiro C, Perez LF, Gude F (2008) Serum TNF-alpha levels in relation to alcohol consumption and common TNF gene polymorphisms. Alcohol 42:513–518. doi:10.1016/j.alcohol.2008.04.008

    Article  CAS  PubMed  Google Scholar 

  29. Yanbaeva DG, Dentener MA, Creutzberg EC, Wesseling G, Wouters EF (2007) Systemic effects of smoking. Chest 131:1557–1566. doi:10.1378/chest.06-2179

    Article  CAS  PubMed  Google Scholar 

  30. Bennet AM, van Maarle MC, Hallqvist J, Morgenstern R, Frostegard J, Wiman B, Prince JA, de Faire U (2006) Association of TNF-alpha serum levels and TNFA promoter polymorphisms with risk of myocardial infarction. Atherosclerosis 187:408–414. doi:10.1016/j.atherosclerosis.2005.09.022

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Anette Baatrup for histology technique support, Yufen Zhang for surgical assistance, and Chen Gan for MRI analysis. We gratefully acknowledge the funding from Velux (25906), Lundbeck, Gigtforeningen Foundation, Aarhus Spine Research Foundation, and the International Cooperation and Natural Science Foundation of Jiangsu province of China (BZ2011046, BL2012069, BK2012490).

Conflict of interest

None.

Author information

Authors and Affiliations

  1. Orthopaedic Research Lab, Aarhus University, Building 1 A, Noerrebrogade 44, 8000, Aarhus C, Denmark

    Ran Kang, Haisheng Li, Kresten Rickers & Cody Bünger

  2. Department of Orthopedic Surgery, Jiangsu Province Hospital on Integration of Chinese and Western Medicine, 100th Shizi Street, Nanjing, 210028, China

    Ran Kang & Lin Xie

  3. The MR Research Centre, Aarhus University Hospital, Skejby, 8000, Aarhus C, Denmark

    Steffen Ringgaard

Authors
  1. Ran Kang
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  2. Haisheng Li
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  5. Lin Xie
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Correspondence to Ran Kang or Lin Xie.

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Kang, R., Li, H., Rickers, K. et al. Intervertebral disc degenerative changes after intradiscal injection of TNF-α in a porcine model. Eur Spine J 24, 2010–2016 (2015). https://doi.org/10.1007/s00586-015-3926-x

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  • DOI: https://doi.org/10.1007/s00586-015-3926-x

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