Skip to main content

Calcium Microcrystal Formation in Recurrent Herniation Patients After Autologous Disc Cell Transplantation

Tissue Engineering and Regenerative Medicine volume 14, pages 803–814 (2017)Cite this article

Abstract

Autologous disc cell transplantation (ADCT) is a cell-based therapy aiming to initiate regeneration of intervertebral disc (IVD) tissue, but little is known about potential risks. This study aims to investigate the presence of structural phenomena accompanying the transformation process after ADCT treatment in IVD disease. Structural phenomena of ADCT-treated patients (Group 1, n = 10) with recurrent disc herniation were compared to conventionally-treated patients with recurrent herniation (Group 2, n = 10) and patients with a first-time herniation (Group 3, n = 10). For ethical reasons, a control group of ADCT patients who did not have a recurrent disc herniation was not possible. Tissue samples were obtained via micro-sequestrectomy after disc herniation and analyzed by micro-computed tomography, scanning electron microscopy, energy dispersive spectroscopy, and histology in terms of calcification zones, tissue structure, cell density, cell morphology, and elemental composition. The major differentiator between sample groups was calcium microcrystal formation in all ADCT samples, not found in any of the control group samples, which may indicate disc degradation. The incorporation of mineral particles provided clear contrast between the different materials and chemical analysis of a single particle indicated the presence of magnesium-containing calcium phosphate. As IVD calcification is a primary indicator of disc degeneration, further investigation of ADCT and detailed investigations assessing each patient’s Pfirrmann degeneration grade following herniation is warranted. Structural phenomena unique to ADCT herniation prompt further investigation of the therapy’s mechanisms and its effect on IVD tissue. However, the impossibility of a perfect control group limits the generalizable interpretation of the results.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. Fanuele JC, Abdu WA, Hanscom B, Weinstein JN. Association between obesity and functional status in patients with spine disease. Spine (Phila Pa 1976). 2002;27:306–12.

    Article  Google Scholar 

  2. Hart LG, Deyo RA, Cherkin DC. Physician office visits for low-back-pain—frequency, clinical-evaluation, and treatment patterns from a US National Survey. Spine. 1995;20:11–9.

    CAS  Article  PubMed  Google Scholar 

  3. Bron JL, Helder MN, Meisel HJ, Van Royen BJ, Smit TH. Repair, regenerative and supportive therapies of the annulus fibrosus: achievements and challenges. Eur Spine J. 2009;18:301–13.

    Article  PubMed  Google Scholar 

  4. Sakai D, Andersson GB. Stem cell therapy for intervertebral disc regeneration: obstacles and solutions. Nat Rev Rheumatol. 2015;11:243–56.

    Article  PubMed  Google Scholar 

  5. Pfirrmann CW, Metzdorf A, Zanetti M, Hodler J, Boos N. Magnetic resonance classification of lumbar intervertebral disc degeneration. Spine (Phila Pa 1976). 2001;26:1873–8.

    CAS  Article  Google Scholar 

  6. Masuda K, Imai Y, Okuma M, Muehleman C, Nakagawa K, Akeda K, et al. 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). 2006;31:742–54.

    Article  Google Scholar 

  7. Zhang Y, An HS, Tannoury C, Thonar EJ, Freedman MK, Anderson DG. Biological treatment for degenerative disc disease. Am J Phys Med Rehabil. 2008;87:694–702.

    Article  PubMed  Google Scholar 

  8. Paesold G, Nerlich AG, Boos N. Biological treatment strategies for disc degeneration: potentials and shortcomings. Eur Spine J. 2007;16:447–68.

    Article  PubMed  Google Scholar 

  9. Ganey T, Hutton WC, Moseley T, Hedrick M, Meisel HJ. Intervertebral disc repair using adipose tissue-derived stem and regenerative cells experiments in a canine model. Spine (Phila Pa 1976). 2009;34:2297–304.

    Article  Google Scholar 

  10. Ganey T, Libera J, Moos V, Alasevic O, Fritsch KG, Meisel HJ, et al. Disc chondrocyte transplantation in a canine model: a treatment for degenerated or damaged intervertebral disc. Spine (Phila Pa 1976). 2003;28:2609–20.

    Article  Google Scholar 

  11. Meisel HJ, Ganey T, Hutton WC, Libera J, Minkus Y, Alasevic O. Clinical experience in cell-based therapeutics: intervention and outcome. Eur Spine J. 2006;15:S397–405.

    Article  PubMed  Google Scholar 

  12. Meisel HJ, Siodla V, Ganey T, Minkus Y, Hutton WC, Alasevic OJ. Clinical experience in cell-based therapeutics: disc chondrocyte transplantation—a treatment for degenerated or damaged intervertebral disc. Biomol Eng. 2007;24:5–21.

    CAS  Article  PubMed  Google Scholar 

  13. Tschugg A, Diepers M, Simone S, Michnacs F, Quirbach S, Strowitzki M, et al. A prospective randomized multicenter phase I/II clinical trial to evaluate safety and efficacy of NOVOCART disk plus autologous disk chondrocyte transplantation in the treatment of nucleotomized and degenerative lumbar disks to avoid secondary disease: safety results of Phase I—a short report. Neurosurg Rev. 2017;40:155–62.

    Article  PubMed  Google Scholar 

  14. Tschugg A, Michnacs F, Strowitzki M, Meisel HJ, Thome C. A prospective multicenter phase I/II clinical trial to evaluate safety and efficacy of NOVOCART Disc plus autologous disc chondrocyte transplantation in the treatment of nucleotomized and degenerative lumbar disc to avoid secondary disease: study protocol for a randomized controlled trial. Trials. 2016;17:108.

    Article  PubMed  PubMed Central  Google Scholar 

  15. Josimovic-Alasevic O, Libera J, Siodla V, Meisel HJ (2006). Verfahren zur Herstellung von Bandscheibenzelltransplantaten und deren Anwendung als Transplantationsmaterial, DE Patent DE10, 2004, 043449, A, 1 09 Mar 2006

  16. Anderson DG, Albert TJ, Fraser JK, Risbud M, Wuisman P, Meisel HJ, et al. Cellular therapy for disc degeneration. Spine. 2005;30:14–9.

    Article  Google Scholar 

  17. Meisel HJ, Ganey T. Nucleus reconstruction by autologous chondrocyte transplantation. In: Mayer HM, editor. Minimally Invasive Spine Surgery. Berlin: Springer; 2006. p. 364–73.

    Chapter  Google Scholar 

  18. Grochulla F, Mayer HM, Korge A. Autologous disc chondrocyte transplantation. In: Mayer HM, editor. Minimally Invasive Spine Surgery. Berlin: Springer; 2006. p. 374–8.

    Chapter  Google Scholar 

  19. Friedmann A, Goehre F, Ludtka C, Mendel T, Meisel HJ, Heilmann A, et al. Microstructure analysis method for evaluating degenerated intervertebral disc tissue. Micron. 2017;92:51–62.

    Article  PubMed  Google Scholar 

  20. Chan WC, Sze KL, Samartzis D, Leung VY, Chan D. Structure and biology of the intervertebral disk in health and disease. Orthop Clin North Am. 2011;42:447–64.

    Article  PubMed  Google Scholar 

  21. Kobayashi S, Meir A, Urban J. Effect of cell density on the rate of glycosaminoglycan accumulation by disc and cartilage cells in vitro. J Orthop Res. 2008;26:493–503.

    CAS  Article  PubMed  Google Scholar 

  22. Adams MA, Roughley PJ. What is intervertebral disc degeneration, and what causes it? Spine. 2006;31:2151–61.

    Article  PubMed  Google Scholar 

  23. Chanchairujira K, Chung CB, Kim JY, Papakonstantinou O, Lee MH, Clopton P, et al. Intervertebral disk calcification of the spine in an elderly population: radiographic prevalence, location, and distribution and correlation with spinal degeneration. Radiology. 2004;230:499–503.

    Article  PubMed  Google Scholar 

  24. Hristova GI, Jarzem P, Ouellet JA, Roughley PJ, Epure LM, Antoniou J, et al. Calcification in human intervertebral disc degeneration and scoliosis. J Orthop Res. 2011;29:1888–95.

    Article  PubMed  Google Scholar 

  25. Lee RS, Kayser MV, Ali SY. Calcium phosphate microcrystal deposition in the human intervertebral disc. J Anat. 2006;208:13–9.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  26. Melrose J, Burkhardt D, Taylor TKF, Dillon CT, Read R, Cake M, et al. Calcification in the ovine intervertebral disc: a model of hydroxyapatite deposition disease. Eur Spine J. 2009;18:479–89.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Meisel H, Ganey T. Minimally invasive spine surgery. In: Mayer HM, editor. Minimally Invasive Spine Surgery, New York: Springer; 2006. p. 364–73.

  28. Cevei M, Roşca E, Liviu L, Muţiu G, Stoicănescu D, Vasile L. Imagistic and histopathologic concordances in degenerative lesions of intervertebral disks. Rom J Morphol Embryol. 2011;52:327–32.

    PubMed  Google Scholar 

  29. Mulisch M, Welsch U, editors. Romeis-Mikroskopische Technik. Berlin: Springer; 2010.

    Google Scholar 

  30. Spurr AR. A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res. 1969;26:31–43.

    CAS  Article  PubMed  Google Scholar 

  31. Reed SJB. Electron Microprobe Analysis and Scanning Electron Microscopy in Geology. 2nd ed. Cambridge: University Press; 2010.

    Google Scholar 

  32. Rutges JP, Jagt van der OP, Oner FC, Verbout AJ, Castelein RJ, Kummer JA, et al. Micro-CT quantification of subchondral endplate changes in intervertebral disc degeneration. Osteoarthr Cartil. 2011;19:89–95.

  33. Weiler C, Schietzsch M, Kirchner T, Nerlich AG, Boos N, Wuertz K. Age-related changes in human cervical, thoracal and lumbar intervertebral disc exhibit a strong intra-individual correlation. Eur Spine J. 2012;21:S810–8.

    Article  PubMed  Google Scholar 

  34. Meredith DS, Huang RC, Nguyen J, Lyman S. Obesity increases the risk of recurrent herniated nucleus pulposus after lumbar microdiscectomy. Spine J. 2010;10:575–80.

    Article  PubMed  Google Scholar 

  35. Miwa S, Yokogawa A, Kobayashi T, Nishimura T, Igarashi K, Inatani H, et al. Risk factors of recurrent lumbar disk herniation a single center study and review of the literature. J Spinal Disord Tech. 2015;28:E265–9.

    Article  PubMed  Google Scholar 

  36. Shamji MF, Bains I, Yong E, Sutherland G, Hurlbert RJ. Treatment of herniated lumbar disk by sequestrectomy or conventional diskectomy. World Neurosurg. 2014;82:879–83.

    Article  PubMed  Google Scholar 

  37. Rutges JP, Duit RA, Kummer JA, Oner FC, van Rijen MH, Verbout AJ, et al. Hypertrophic differentiation and calcification degeneration. Osteoarthr Cartil. 2010;18:1487–95.

    CAS  Article  PubMed  Google Scholar 

  38. Vo N, Niedernhofer LJ, Nasto LA, Jacobs L, Robbins PD, Kang J, et al. An overview of underlying causes and animal models for the study of age-related degenerative disorders of the spine and synovial joints. J Orthop Res. 2013;31:831–7.

    Article  PubMed  PubMed Central  Google Scholar 

  39. Weiler C. In situ analysis of pathomechanisms of human intervertebral disc degeneration. Pathologe. 2013;34:251–9.

    Article  PubMed  Google Scholar 

  40. Weiler C, Lopez-Ramos M, Mayer HM, Korge A, Siepe CJ, Wuertz K, et al. Histological analysis of surgical lumbar intervertebral disc tissue provides evidence for an association between disc degeneration and increased body mass index. BMC Res Notes. 2011;4:497.

    Article  PubMed  PubMed Central  Google Scholar 

  41. Boos N, Weissbach S, Rohrbach H, Weiler C, Spratt KF, Nerlich AG. Classification of age-related changes in lumbar intervertebral discs. Spine. 2002;27:2631–44.

    Article  PubMed  Google Scholar 

  42. Johnson WEB, Eisenstein SM, Roberts S. Cell cluster formation in degenerate lumbar intervertebral discs is associated with increased disc cell proliferation. Connect Tissue Res. 2001;42:197–207.

    CAS  Article  PubMed  Google Scholar 

  43. Sharp CA, Roberts S, Evans H, Brown SJ. Disc cell clusters in pathological human intervertebral discs are associated with increased stress protein immunostaining. Eur Spine J. 2009;18:1587–94.

    Article  PubMed  PubMed Central  Google Scholar 

  44. Ferguson SJ, Ito K, Nolte LP. Fluid flow and convective transport of solutes within the intervertebral disc. J Biomech. 2004;37:213–21.

    Article  PubMed  Google Scholar 

  45. Huang CY, Gu WY. Effects of mechanical compression on metabolism and distribution of oxygen and lactate in intervertebral disc. J Biomech. 2008;41:1184–96.

    Article  PubMed  PubMed Central  Google Scholar 

  46. Soukane DM, Shirazi-Adl A, Urban JP. Computation of coupled diffusion of oxygen, glucose and lactic acid in an intervertebral disc. J Biomech. 2007;40:2645–54.

    Article  PubMed  Google Scholar 

  47. Bonewald LF, Harris SE, Rosser J, Dallas MR, Dallas SL, Camacho NP, et al. von Kossa staining alone is not sufficient to confirm that mineralization in vitro represents bone formation. Calcif Tissue Int. 2003;72:537–47.

    CAS  Article  PubMed  Google Scholar 

  48. Halverson PB. Calcium crystal-associated diseases. Curr Opin Rheumatol. 1996;8:259–61.

    CAS  Article  PubMed  Google Scholar 

  49. Lagier R, Baud CA. Magnesium whitlockite, a calcium phosphate crystal of special interest in pathology. Pathol Res Pract. 2003;199:329–35.

    CAS  Article  PubMed  Google Scholar 

  50. Feinberg J, Boachie-Adjei O, Bullough PG, Boskey AL. The distribution of calcific deposits in intervertebral disks of the lumbosacral spine. Clin Orthop Relat Res. 1990;254:303–10.

    Google Scholar 

  51. Tsutsumi S, Yasumoto Y, Ito M. Idiopathic intervertebral disk calcification in childhood: a case report and review of literature. Childs Nerv. Syst. 2011;27:1045–51.

    Article  PubMed  Google Scholar 

  52. Zhao CQ, Wang LM, Jiang LS, Dai LY. The cell biology of intervertebral disc aging and degeneration. Ageing Res Rev. 2007;6:247–61.

    Article  PubMed  Google Scholar 

  53. Liu LT, Huang B, Li CQ, Zhuang Y, Wang J, Zhou Y. Characteristics of stem cells derived from the degenerated human intervertebral disc cartilage endplate. PLoS One. 2011;6:e26285.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  54. Kluba T, Niemeyer T, Gaissmaier C, Grunder T. Human anulus fibrosis and nucleus pulposus cells of the intervertebral disc—effect of degeneration and culture system on cell phenotype. Spine. 2005;30:2743–8.

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

The authors would like to thank U. Heunemann, Nico Teuscher, and A. Götz for technical assistance. We also thank Dr. Uwe Spohn and Andreas Cismak for scientific discussions and technical help. We extend special thanks to all the patients who participated in this study. The work presented in this paper was made possible by funding from the German Federal Ministry of Education and Research (BMBF, 1315883), the DAAD RISE internship program, and the Whitaker Biomedical Engineering Research Fellowship.

Author information

Affiliations

  1. Translational Centre of Regenerative Medicine TRM, University of Leipzig, Philipp-Rosenthal-Straße 55, 04103, Leipzig, Germany

    S. Schwan, C. Ludtka, A. Friedmann & F. Goehre

  2. Department of Biological and Macromolecular Materials, Fraunhofer Institute for Microstructure of Materials and Systems IMWS, Walter-Huelse-Str. 1, 06120, Halle (Saale), Germany

    S. Schwan, C. Ludtka, A. Friedmann & A. Heilmann

  3. Department of Chemical and Biomolecular Engineering, University of Tennessee, 1512 Middle Drive, Knoxville, TN, 37996, USA

    C. Ludtka

  4. Department of Trauma Surgery, Friedrich-Schiller-University, Am Klinikum 1, 07747, Jena, Germany

    T. Mendel

  5. Department of Trauma Surgery, BG-Klinikum Bergmannstrost Halle, Merseburger Straße 165, 06112, Halle (Saale), Germany

    T. Mendel

  6. Department of Diagnostic Imaging and Interventional Radiology, BG-Klinikum Bergmannstrost Halle, Merseburger Straße 165, 06112, Halle (Saale), Germany

    I. Kaden

  7. Department of Neurosurgery, BG-Klinikum Bergmannstrost Halle, Merseburger Straße 165, 06112, Halle (Saale), Germany

    H. J. Meisel & F. Goehre

  8. Department of Neurosurgery, Helsinki University Central Hospital, University of Helsinki, Topeliuksenkatu 5, 00029, Helsinki, Finland

    F. Goehre

Authors
  1. S. Schwan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C. Ludtka
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. Friedmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. T. Mendel
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H. J. Meisel
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A. Heilmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. I. Kaden
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. F. Goehre
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to S. Schwan.

Ethics declarations

Conflict of interest

This work was supported by the German Federal Ministry of Education and Research (BMBF, PtJ-Bio, 0315883) and the DAAD RISE internship program, and the Whitaker Biomedical Engineering Research Fellowship.

Ethical review committee

The study was approved by the independent medical ethical committee as well as the scientific committee from the faculty of medicine Martin-Luther University Halle-Wittenberg and confirmed at 14 July 2010 under approval no. EK-MLU/14072010/hm-bü.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Schwan, S., Ludtka, C., Friedmann, A. et al. Calcium Microcrystal Formation in Recurrent Herniation Patients After Autologous Disc Cell Transplantation. Tissue Eng Regen Med 14, 803–814 (2017). https://doi.org/10.1007/s13770-017-0076-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13770-017-0076-8

Keywords

  • Recurrent disc herniation
  • Regenerative therapy
  • Disc calcification
  • Intervertebral disc