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01-05-2012 | Original Article

Ex vivo observation of human intervertebral disc tissue and cells isolated from degenerated intervertebral discs

Authors: Gabriela Ciapetti, Donatella Granchi, Valentina Devescovi, Elisa Leonardi, Tiziana Greggi, Mario Di Silvestre, Nicola Baldini

Published in: European Spine Journal | Special Issue 1/2012

Abstract

Purpose

Disc degeneration, and associated low back pain, are a primary cause of disability. Disc degeneration is characterized by dysfunctional cells and loss of proteoglycans: since intervertebral tissue has a limited capacity to regenerate, this process is at present considered irreversible. Recently, cell therapy has been suggested to provide more successful treatment of IVD degeneration. To understand the potential of cells to restore IVD structure/function, tissue samples from degenerated IVD versus healthy discs have been compared.

Methods

Discal tissue from 27 patients (40.17 ± 11 years) undergoing surgery for degenerative disc disease (DDD), DDD + herniation and congenital scoliosis, as controls, was investigated. Cells and matrix in the nucleus pulposus (NP) and annulus fibrosus (AF) were characterized by histology. AF- and NP-derived cells were isolated, expanded and characterized for senescence and gene expression. Three-dimensional NP pellets were cultured and stained for glycosaminoglycan formation.

Results

Phenotypical markers of degeneration, such as cell clusters, chondrons, and collagen disorganization were seen in the degenerate samples. In severe degeneration, granulation tissue and peripheral vascularization were observed. No correlation was found between the Pfirrmann clinical score and the extent of degeneration.

Conclusion

The tissue disorganization in degenerate discs and the paucity of cells out of cluster/chondron association, make the IVD-derived cells an unreliable option for disc regeneration.

Miller JA, Schmatz M, Schultz AB (1988) Lumbar disc degeneration: correlation with age, sex, and spine level in 600 autopsy specimens. Spine 13:173–178PubMedCrossRef

Hadjipavlou AG, Tzermiadianos MN, Bogduk N, Zindrick MR (2008) The pathophysiology of disc degeneration: a critical review. J Bone Jt Surg Br 90:1261–1270CrossRef

Anderson DG, Risbud MV, Shapiro IM, Vaccaro AR, Albert TJ (2005) Cell-based therapy for disc repair. Spine J 5:297S–303SPubMedCrossRef

Nerurkar NL, Elliot DM, Mauck RL (2010) Mechanical design criteria for intervertebral disc tissue engineering. J Biomech 43(6):1017–1030PubMedCrossRef

Roberts S, Evans H, Trived J, Menage J (2006) Histology and pathology of the human intervertebral disc. J Bone Joint Surg A 88(S2):10–14CrossRef

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. J Clin Pathol: Mol Pathol 55:91–97CrossRef

Richardson SM, Mobasheri A, Freemont AJ, Hoyland JA (2007) Intervertebral disc biology, degeneration and novel tissue engineering and regenerative medicine therapies. Histol Histopathol 22:1033–1041PubMed

Bertolo A, Thiede T, Aebli N, Baur M, Ferguson SJ, Stoyanov JV (2011) Human mesenchymal stem cell co-culture modulates the immunological properties of human intervertebral disc tissue fragments in vitro. Eur Spine J 20(4):592–603PubMedCrossRef

Vadalà G, Sowa G, Hubert M, Gilbertson LG, Denaro V, Kang JD (2011) Mesenchymal stem cells injection in degenerated intervertebral disc: cell leakage may induce osteophyte formation. J Tissue Eng Regen Med 13. doi:10.1002/term.433

10.

Ford JL, Jones P, Downes S (2002) Cellularity of human annulus tissue: an investigation into the cellularity of tissue of different pathologies. Histopathology 41:531–537PubMedCrossRef

11.

Debacq-Chainiaux F, Erusalimsky JD, Campisi J, Toussaint O (2009) Protocols to detect senescence-associated beta-galactosidase (SA-βgal) activity, a biomarker of senescent cells in culture and in vivo. Nat Protoc 4(12):1798–1806PubMedCrossRef

12.

Mouritzen P, Noerholm M, Nielsen PS, Jacobsen N, Lomholt C, Pfundheller HM, Tolstrup N (2005) Probe library: a new method for faster design and execution of quantitative real-time PCR. Nat Methods 4:313–316CrossRef

13.

Johnson WE, Eisenstein SM, Roberts S (2001) Cell cluster formation in degenerate lumbar intervertebral discs is associated with increased disc cell proliferation. Connect Tissue Res 42(3):197–207PubMedCrossRef

14.

Livshits G, Popham M, Malkin I, Sambrook PN, Macgregor AJ, Spector T, Williams FM (2011) Lumbar disc degeneration and genetic factors are the main risk factors for low back pain in women: the UK Twin Spine Study. Ann Rheum Dis 70(10):1740–1745PubMedCrossRef

15.

Kepler CK, Anderson DG, Tannoury C, Ponnappan RK (2011) Intervertebral disk degeneration and emerging biologic treatments. J Am Acad Orthop Surg 19(9):543–553PubMed

16.

Peng B, Hao J, Hou S, Wu W, Jiang D, Fu X, Yang Y (2006) Possible pathogenesis of painful intervertebral disc degeneration. Spine (Phila Pa 1976) 31(5):560–566CrossRef

17.

Stephan S, Johnson WE, Roberts S (2011) The influence of nutrient supply and cell density on the growth and survival of intervertebral disc cells in 3D culture. Eur Cell Mater 22:97–108PubMed

18.

Chou AI, Bansal A, Miller GJ, Nicoll SB (2006) The effect of serial monolayer passaging on the collagen expression profile of outer and inner anulus fibrosus cells. Spine (Phila Pa 1976) 31(17):1875–1881CrossRef

19.

Gruber HE, Hoelscher GL, Ingram JA, Zinchenko N, Hanley EN Jr (2010) Senescent vs. non-senescent cells in the human annulus in vivo: cell harvest with laser capture microdissection and gene expression studies with microarray analysis. BMC Biotechnol 10:1–10CrossRef

20.

Yang Z, Huang CY, Candiotti KA, Zeng X, Yuan T, Li J, Yu H, Abdi S (2011) Sox-9 facilitates differentiation of adipose tissue-derived stem cells into a chondrocyte-like phenotype in vitro. J Orthop Res 29:1291–1297PubMedCrossRef

21.

Zhao CQ, Wang LM, Jiang LS, Dai LY (2007) The cell biology of intervertebral disc aging and degeneration. Ageing Res Rev 6:247–261PubMedCrossRef

22.

Preradovic A, Kleinpeter G, Feichtinger H, Balaun E, Krugluger W (2005) Quantitation of collagen I, collagen II and aggrecan mRNA and expression of the corresponding proteins in human nucleus pulposus cells in monolayer cultures. Cell Tissue Res 321(3):459–464PubMedCrossRef

23.

Strassburg S, Richardson SM, Freemont AJ, Hoyland JA (2010) Co-culture induces mesenchymal stem cell differentiation and modulation of the degenerate human nucleus pulposus cell phenotype. Regen Med 5(5):701–711PubMedCrossRef

24.

Le Maitre CL, Baird P, Freemont AJ, Hoyland JA (2009) An in vitro study investigating the survival and phenotype of mesenchymal stem cells following injection into nucleus pulposus tissue. Arthritis Res Ther 11(1):R20PubMedCrossRef

25.

Bron JL, Helder MN, Meisel HJ, Van Royen BJ, Smit TH (2009) Repair, regenerative and supportive therapies of the annulus fibrosus: achievements and challenges. Eur Spine J 18(3):301–313PubMedCrossRef

Title: Ex vivo observation of human intervertebral disc tissue and cells isolated from degenerated intervertebral discs
Authors: Gabriela Ciapetti
Donatella Granchi
Valentina Devescovi
Elisa Leonardi
Tiziana Greggi
Mario Di Silvestre
Nicola Baldini
Publication date: 01-05-2012
Publisher: Springer-Verlag
Published in: European Spine Journal / Issue Special Issue 1/2012
Print ISSN: 0940-6719
Electronic ISSN: 1432-0932
DOI: https://doi.org/10.1007/s00586-012-2234-y

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Ex vivo observation of human intervertebral disc tissue and cells isolated from degenerated intervertebral discs

Abstract

Purpose

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

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