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A Biodegradable Polymeric Matrix for the Repair of Annulus Fibrosus Defects in Intervertebral Discs

Abstract

Background:

Tissue defects in the annulus fibrosus (AF) due to intervertebral disc (IVD) degeneration or after nucleodiscectomy have little self-healing capacity. To prevent progressive degeneration of the IVD, the AF must be repaired. Biological closure has not yet been achieved and is a challenge for the research community. In this study, a scaffold made of absorbable poly (glycolic acid) (PGA) and hyaluronan (HA) that exhibit excellent biocompatibility and cell colonization properties was used to repair AF defects in an ovine model.

Methods:

A partial resection was performed in AF in L3/4 or L4/5 of 10 sheep and PGA-HA scaffolds were implanted on the defects (n = 5), while defects in the control group were left untreated (n = 5). Three months post-operation, the lumbar discs were sectioned and stained with hematoxylin and eosin and safranin-O/fast-green. Histological features including proteoglycan content, annular structure, cellular morphology, blood vessel ingrowth and tear/cleft formation were scored using a modified scoring scheme by 3 investigators and evaluated by a pathologist independently.

Results:

The treated AF exhibited significantly enhanced repair tissue structure with signs of proteoglycan formation compared to the untreated group. The median scores were 4.3 for the treated and 9.8 for the untreated group. Cystic degeneration, perivascular infiltration, inflammation and necrosis were only present in the untreated group. Blood vessel ingrowth and tear/cleft formation were increased, though not significant, in the untreated group while cell morphology was comparable in both groups.

Conclusion:

PGA-HA scaffolds used for AF closure support repair tissue formation in an ovine lumbar disc defect model.

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References

  1. Harmon MD, Ramos DM, Nithyadevi D, Bordett R, Rudraiah S, Nukavarapu SP, et al. Growing a backbone - functional biomaterials and structures for intervertebral disc (IVD) repair and regeneration: challenges, innovations, and future directions. Biomater Sci. 2020;8:1216–39.

    CAS  Article  PubMed  Google Scholar 

  2. Shu CC, Smith MM, Smith SM, Dart AJ, Little CB, Melrose J. A histopathological scheme for the quantitative scoring of intervertebral disc degeneration and the therapeutic utility of adult mesenchymal stem cells for intervertebral disc regeneration. Int J Mol Sci. 2017;18:1049.

    Article  PubMed Central  Google Scholar 

  3. Cabraja M, Endres M, Hegewald AA, Vetterlein S, Thomé C, Woiciechowsky C, et al. A 3D environment for anulus fibrosus regeneration. J Neurosurg Spine. 2012;17:177–83.

    Article  PubMed  Google Scholar 

  4. Sharifi S, Bulstra SK, Grijpma DW, Kuijer R. Treatment of the degenerated intervertebral disc; closure, repair and regeneration of the annulus fibrosus. J Tissue Eng Regen Med. 2015;9:1120–32.

    Article  PubMed  Google Scholar 

  5. Tavakoli J, Diwan AD, Tipper JL. Advanced strategies for the regeneration of lumbar disc annulus fibrosus. Int J Mol Sci. 2020;21:4889.

    Article  PubMed Central  Google Scholar 

  6. Alini M, Li W, Markovic P, Aebi M, Spiro RC, Roughley PJ. The potential and limitations of a cell-seeded collagen/hyaluronan scaffold to engineer an intervertebral disc-like matrix. Spine (Phila Pa 1976). 2003;28:446–53.

    Google Scholar 

  7. Park SH, Gil ES, Cho H, Mandal BB, Tien LW, Min BH, et al. Intervertebral disk tissue engineering using biphasic silk composite scaffolds. Tissue Eng Part A. 2012;18:447–58.

    CAS  Article  PubMed  Google Scholar 

  8. Shao X, Hunter CJ. Developing an alginate/chitosan hybrid fiber scaffold for annulus fibrosus cells. J Biomed Mater Res A. 2007;82:701–10.

    Article  Google Scholar 

  9. Shen B, Wei A, Bhargav D, Kishen T, Diwan AD. Hyaluronan: its potential application in intervertebral disc regeneration. Orthop Res Rev. 2010;2:17–26.

    CAS  Article  Google Scholar 

  10. Chou AI, Reza AT, Nicoll SB. Distinct intervertebral disc cell populations adopt similar phenotypes in three-dimensional culture. Tissue Eng Part A. 2008;14:2079–87.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  11. Koepsell L, Remund T, Bao J, Neufeld D, Fong H, Deng Y. Tissue engineering of annulus fibrosus using electrospun fibrous scaffolds with aligned polycaprolactone fibers. J Biomed Mater Res Part A. 2011;99:564–75.

    Article  Google Scholar 

  12. Lei Z, Song Z, Bao-chang C. Actuality and challenge of biomaterials in annulus fibrosus repair. Chin J Tissue Eng Res. 2018;22:958–63.

    Google Scholar 

  13. Moran JM, Pazzano D, Bonassar LJ. Characterization of polylactic acid–polyglycolic acid composites for cartilage tissue engineering. Tissue Eng. 2003;9:63–70.

    CAS  Article  PubMed  Google Scholar 

  14. Abbushi A, Endres M, Cabraja M, Kroppenstedt SN, Thomale UW, Sittinger M, et al. Regeneration of intervertebral disc tissue by resorbable cell-free polyglycolic acid-based implants in a rabbit model of disc degeneration. Spine (Phila Pa 1976). 2008;33:1527–32.

    Article  Google Scholar 

  15. Endres M, Abbushi A, Thomale UW, Cabraja M, Kroppenstedt SN, Morawietz L, et al. Intervertebral disc regeneration after implantation of a cell-free bioresorbable implant in a rabbit disc degeneration model. Biomaterials. 2010;31:5836–41.

    CAS  Article  PubMed  Google Scholar 

  16. Hegewald AA, Medved F, Feng D, Tsagogiorgas C, Beierfuß A, Schindler GAK, et al. Enhancing tissue repair in annulus fibrosus defects of the intervertebral disc: analysis of a bio-integrative annulus implant in an in-vivo ovine model. J Tissue Eng Regen Med. 2015;9:405–14.

    Article  PubMed  Google Scholar 

  17. Hegewald AA, Enz A, Endres M, Sittinger M, Woiciechowsky C, Thomé C, et al. Engineering of polymer-based grafts with cells derived from human nucleus pulposus tissue of the lumbar spine. J Tissue Eng Regen Med. 2011;5:275–82.

    CAS  Article  PubMed  Google Scholar 

  18. Siclari A, Mascaro G, Kaps C, Boux E. A 5-year follow-up after cartilage repair in the knee using a platelet-rich plasma-immersed polymer-based implant. Open Orthop J. 2014;8:346–54.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Bergmanna A, Hans Peter Abt. 7 months clinical follow-up after arthrotomic implantation of chondrotissue in a cartilage lesion of the lateral talus. Fuß & Sprunggelenk. 2009;8:264–7.

  20. Woiciechowsky C, Abbushi A, Zenclussen ML, Casalis P, Krüger JP, Freymann U, et al. Regeneration of nucleus pulposus tissue in an ovine intervertebral disc degeneration model by cell-free resorbable polymer scaffolds. J Tissue Eng Regen Med. 2014;8:811–20.

    CAS  Article  PubMed  Google Scholar 

  21. Hondke S, Cabraja M, Krüger JP, Stich S, Hartwig T, Sittinger M, et al. Proliferation, migration, and ECM formation potential of human annulus fibrosus cells is independent of degeneration status. Cartilage. 2020;11:192–202.

    CAS  Article  PubMed  Google Scholar 

  22. Boos N, Weissbach S, Rohrbach H, Weiler C, Spratt KF, Nerlich AG. Classification of age-related changes in lumbar intervertebral discs: 2002 Volvo Award in basic science. Spine (Phila Pa 1976). 2002;27:2631–44.

    Article  Google Scholar 

  23. Rutges JP, Duit RA, Kummer JA, Bekkers JE, Oner FC, Castelein RM, et al. A validated new histological classification for intervertebral disc degeneration. Osteoarthritis Cartilage. 2013;21:2039–47.

    CAS  Article  Google Scholar 

  24. Ostach PKS. Model for the implantation of implants stimulating the body’s own mechanisms for repair and regeneration of ovine annulus fibrosus defects. [Doctoral dissertation]. Free University Berlin; 2019.

  25. Koo TK, Li MY. Cracking the code: providing insight into the fundamentals of research and evidence-based practice a guideline of selecting and reporting intraclass correlation coefficients for reliability research. J Chiropr Med. 2016;15:155–63.

    Article  PubMed  PubMed Central  Google Scholar 

  26. Lee SH, Kim BS, Kim SH, Choi SW, Jeong SI, Kwon IK, et al. Elastic biodegradable poly(glycolide-co-caprolactone) scaffold for tissue engineering. J Biomed Mater Res A. 2003;66:29–37.

    Article  Google Scholar 

  27. Lyons G, Eisenstein SM, Sweet MB. Biochemical changes in intervertebral disc degeneration. Biochim Biophys Acta. 1981;673:443–53.

    CAS  Article  Google Scholar 

  28. Papakonstantinou E, Roth M, Karakiulakis G. Hyaluronic acid: A key molecule in skin aging. Dermatoendocrinol. 2012;4:253–8.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  29. Urban JP, Smith S, Fairbank JC. Nutrition of the Intervertebral disc. Spine (Phila Pa 1976). 2004;29:2700–9.

    Article  PubMed  Google Scholar 

  30. 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 

  31. Melrose J, Roberts S, Smith S, Menage J, Ghosh P. Increased nerve and blood vessel ingrowth associated with proteoglycan depletion in an ovine anular lesion model of experimental disc degeneration. Spine (Phila Pa 1976). 2002;27:1278–85.

    Article  PubMed  Google Scholar 

  32. Melrose J. Repair of annular disc lesions: spatial and temporal localisation of TGF-beta, SPARC (osteonectin/BM-40) and FGF-2 are early responses in experimentally injured ovine discs. Trans Orthop Res Soc. 2001;26:896.

    Google Scholar 

  33. Sartoretto SC, Uzeda MJ, Miguel FB, Nascimento JR, Ascoli F, Calasans-Maia MD. Sheep as an experimental model for biomaterial implant evaluation. Acta Ortop Bras. 2016;24:262–6.

    Article  PubMed  PubMed Central  Google Scholar 

  34. Smit TH. The use of a quadruped as an in vivo model for the study of the spine - biomechanical considerations. Eur Spine J. 2002;11:137–44.

    Article  PubMed  PubMed Central  Google Scholar 

  35. Bailey AS, Adler F, Min Lai S, Asher MA. A comparison between bipedal and quadrupedal rats: do bipedal rats actually assume an upright posture? Spine (Phila Pa 1976). 2001;26:308–13.

    Article  Google Scholar 

  36. Alini M, Eisenstein SM, Ito K, Little C, Kettler AA, Masuda K, et al. Are animal models useful for studying human disc disorders/degeneration? Eur Spine J. 2008;17:2–19.

    Article  PubMed  Google Scholar 

  37. Bouhsina N, Decante C, Hardel J, B, Madec S, Abadie J, Hamel A, et al. Correlation between magnetic resonance, X-ray imaging alterations and histological changes in an ovine model of age-related disc degeneration. Eur Cells Mater. 2021;42:166–78.

  38. Kruger JP, Ketzmar AK, Endres M, Pruss A, Siclari A, Kaps C. Human platelet-rich plasma induces chondrogenic differentiation of subchondral progenitor cells in polyglycolic acid-hyaluronan scaffolds. J Biomed Mater Res Part B Appl Biomater. 2014;102:681–92.

    Article  Google Scholar 

  39. Yang D, Lu X, Hong Y, Xi T, Zhang D. The molecular mechanism of mediation of adsorbed serum proteins to endothelial cells adhesion and growth on biomaterials. Biomaterials. 2013;34:5747–58.

    CAS  Article  PubMed  Google Scholar 

  40. Wilson CJ, Clegg RE, Leavesley DI, Pearcy MJ. Mediation of biomaterial-cell interactions by adsorbed proteins: a review. Tissue Eng. 2005;11:1–18.

    CAS  Article  PubMed  Google Scholar 

  41. Allen LT, Tosetto M, Miller IS, O’Connor DP, Penney SC, Lynch I, et al. Surface-induced changes in protein adsorption and implications for cellular phenotypic responses to surface interaction. Biomaterials. 2006;27:3096–108.

    CAS  Article  PubMed  Google Scholar 

  42. Kalwitz G, Andreas K, Endres M, Neumann K, Notter M, Ringe J, et al. Chemokine profile of human serum from whole blood: migratory effects of CXCL-10 and CXCL-11 on human mesenchymal stem cells. Connect Tissue Res. 2010;51:113–22.

    CAS  Article  PubMed  Google Scholar 

  43. Hegewald AA, Knecht S, Baumgartner D, Gerber H, Endres M, Kaps C, et al. Biomechanical testing of a polymer-based biomaterial for the restoration of spinal stability after nucleotomy. J Orthop Surg Res. 2009;4:25.

    Article  Google Scholar 

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Acknowledgements

The authors thank Dr. Katja Reiter, Dr. Pia Ostach and Prof. Dr. Christian Große-Siestrup for animal surgery. We also like to thank the Research facilities for experimental medicine (FEM) and the animal holding unit of the Charité-Universitaetsmedizin Berlin for animal care. Grateful thanks to Samuel Vetterlein and Anja Wachtel for technical support of sample preparation and histochemical staining. The research leading to these results received funding from Federal Ministry of Education and Research (BMBF) grant no. 13N13435 and 13N13437.

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Correspondence to Michaela Endres.

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Conflict of interest

MRSF, JPK, PB and ME are employees of the research company TransTissue Technologies GmbH and receive salaries from the company. All other declare no conflict of interest.

Ethical statement

All procedures performed in this study involving animals were approved by the State Office for Health and Social Affairs in Berlin, Germany (LaGeSo No. G 0093/16). The study was performed under German Animal Welfare Act (§ 8.1 and § 7.2).

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Saghari Fard, M.R., Krueger, J.P., Stich, S. et al. A Biodegradable Polymeric Matrix for the Repair of Annulus Fibrosus Defects in Intervertebral Discs. Tissue Eng Regen Med (2022). https://doi.org/10.1007/s13770-022-00466-0

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  • DOI: https://doi.org/10.1007/s13770-022-00466-0

Keywords

  • Annulus fibrosus
  • PGA-HA
  • Ovine
  • Intervertebral discs