European Spine Journal

, Volume 28, Issue 4, pp 633–648 | Cite as

A histocytological and radiological overview of the natural history of intervertebral disk: from embryonic formation to age-related degeneration

  • Feng Wang
  • Cong Zhang
  • Arjun Sinkemani
  • Rui Shi
  • Zhi-Yang Xie
  • Lu Chen
  • Lu Mao
  • Xiao-Tao WuEmail author
Original Article



To elucidate the natural history of intervertebral disk (IVD) and characterize its embryonic beginnings and age-related degeneration.


Coronal sections of embryonic (E13.5-neonatal) and postnatal (4–60-week-old) Sprague–Dawley rat IVD were stained by a series of histological stainings (hematoxylin and eosin, Alcian blue, Picrosirius red, Masson, Periodic acid–Schiff). Growth kinetics within embryonic IVD were evaluated by immunohistochemical staining of Ki67 and proliferating cell nuclear antigen. Postnatal maturation and degeneration of IVD were visualized on radiology by X-ray, CT, and MR imaging.


During the formation of rat IVD, inner annulus fibrosus (AF) and cartilaginous endplate (CEP) shared similar cell density, extracellular matrix, and potential of growth kinetics; notochord provided increased and enlarged cytoplasmic vacuoles to generate nucleus pulposus (NP), part of which was retained within CEP. Postnatally, vacuolated notochord cells were reduced by devacuolation, while chondrocytic NP cells increased; cartilaginous layers of CEP were narrowed by vertebrae growth and secondary ossification; fibrotic portion of AF decreased as cartilaginous matrix accumulated and infiltrated outward. In aged and degenerated IVD, large longitudinal fissures were detected near the boundaries between inner and outer AF, whereas both reduced cellularity and accumulated cell clusters were evident within the dehydrated NP; only part of these histocytological changes could be reported on radiology.


By showing that the natural history of IVD is orchestrated by a dynamic histocytological regulation, our study may facilitate better understanding of the developmental defects, cellular heterogeneity, age-related degenerative mechanisms, and biological regeneration of IVD.

Graphical abstract

These slides can be retrieved under Electronic Supplementary Material.


Intervertebral Disk Embryogenesis Cytoplasmic Vacuole Notochord Sclerotome 



This study was supported by the National Natural Science Foundation of China (No. 81201423, No. 81272035, No. 81572170) and the Fundamental Research Funds of the Central Universities (No. 2242017K3DN06). The authors would like to acknowledge He-Ling Fu, Yuan Zheng, and Dan Bao, from the Animal Core facility of Nanjing Medical University, for their aids in radiological imaging and analysis.

Compliance with ethical standards

Conflict of interest


Supplementary material

586_2019_5903_MOESM1_ESM.pptx (1.7 mb)
Supplementary material 1 (PPTX 1786 kb)


  1. 1.
    Risbud MV, Shapiro IM (2014) Role of cytokines in intervertebral disc degeneration: pain and disc content. Nat Rev Rheumatol 10(1):44–56PubMedGoogle Scholar
  2. 2.
    Wang F, Shi R, Cai F, Wang YT, Wu XT (2015) Stem cell approaches to intervertebral disc regeneration: obstacles from the disc microenvironment. Stem Cells Dev 24(21):2479–2495PubMedGoogle Scholar
  3. 3.
    Wang F, Cai F, Shi R, Wang XH, Wu XT (2016) Aging and age related stresses: a senescence mechanism of intervertebral disc degeneration. Osteoarthritis Cartilage 24(3):398–408PubMedGoogle Scholar
  4. 4.
    Vos T, Flaxman AD, Naghavi M, Lozano R et al (2012) Years lived with disability (YLDs) for 1160 sequelae of 289 diseases and injuries 1990–2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet 380(9859):2163–2196PubMedPubMedCentralGoogle Scholar
  5. 5.
    Wu X, Zhuang S, Mao Z, Chen H (2006) Microendoscopic discectomy for lumbar disc herniation: surgical technique and outcome in 873 consecutive cases. Spine 31(23):2689–2694PubMedGoogle Scholar
  6. 6.
    Sinkemani A, Hong X, Gao ZX, Zhuang SY et al (2015) Outcomes of microendoscopic discectomy and percutaneous transforaminal endoscopic discectomy for the treatment of lumbar disc herniation: a comparative retrospective study. Asian Spine J 9(6):833–840PubMedPubMedCentralGoogle Scholar
  7. 7.
    Wang K, Hong X, Zhou BY, Bao JP et al (2015) Evaluation of transforaminal endoscopic lumbar discectomy in the treatment of lumbar disc herniation. Int Orthop 39(8):1599–1604PubMedGoogle Scholar
  8. 8.
    Luk KD, Samartzis D (2015) Intervertebral disc “dysgeneration”. Spine J 15(9):1915–1918PubMedGoogle Scholar
  9. 9.
    Pfirrmann CW, Metzdorf A, Zanetti M, Hodler J, Boos N (2001) Magnetic resonance classification of lumbar intervertebral disc degeneration. Spine 26(17):1873–1878PubMedGoogle Scholar
  10. 10.
    Davies BM, Atkinson RA, Ludwinski F, Freemont AJ et al (2016) Qualitative grading of disc degeneration by magnetic resonance in the lumbar and cervical spine: lack of correlation with histology in surgical cases. Br J Neurosurg 30(4):414–421PubMedGoogle Scholar
  11. 11.
    Battie MC, Videman T, Levalahti E, Gill K, Kaprio J (2007) Heritability of low back pain and the role of disc degeneration. Pain 131(3):272–280PubMedGoogle Scholar
  12. 12.
    Samartzis D, Karppinen J, Mok F, Fong DY, Luk KD, Cheung KM (2011) A population-based study of juvenile disc degeneration and its association with overweight and obesity, low back pain, and diminished functional status. J Bone Joint Surg Am 93(7):662–670PubMedGoogle Scholar
  13. 13.
    Cheung KM, Samartzis D, Karppinen J, Mok FP et al (2010) Intervertebral disc degeneration: new insights based on “skipped” level disc pathology. Arthritis Rheum 62(8):2392–2400PubMedGoogle Scholar
  14. 14.
    Cheung KM, Karppinen J, Chan D, Ho DW et al (2009) Prevalence and pattern of lumbar magnetic resonance imaging changes in a population study of one thousand forty-three individuals. Spine 34(9):934–940PubMedGoogle Scholar
  15. 15.
    Scaal M (2016) Early development of the vertebral column. Semin Cell Dev Biol 49:83–91PubMedGoogle Scholar
  16. 16.
    Corallo D, Trapani V, Bonaldo P (2015) The notochord: structure and functions. Cell Mol Life Sci 72(16):2989–3008PubMedGoogle Scholar
  17. 17.
    Chan WC, Au TY, Tam V, Cheah KS, Chan D (2014) Coming together is a beginning: the making of an intervertebral disc. Birth Defects Res C Embryo Today 102(1):83–100PubMedGoogle Scholar
  18. 18.
    Rodrigues-Pinto R, Richardson SM, Hoyland JA (2014) An understanding of intervertebral disc development, maturation and cell phenotype provides clues to direct cell-based tissue regeneration therapies for disc degeneration. Eur Spine J 23(9):1803–1814PubMedGoogle Scholar
  19. 19.
    Risbud MV, Shapiro IM (2011) Notochordal cells in the adult intervertebral disc: new perspective on an old question. Crit Rev Eukaryot Gene Expr 21(1):29–41PubMedPubMedCentralGoogle Scholar
  20. 20.
    Risbud MV, Schaer TP, Shapiro IM (2010) Toward an understanding of the role of notochordal cells in the adult intervertebral disc: from discord to accord. Dev Dyn 239(8):2141–2148PubMedPubMedCentralGoogle Scholar
  21. 21.
    Fleming A, Keynes R, Tannahill D (2004) A central role for the notochord in vertebral patterning. Development 131(4):873–880PubMedGoogle Scholar
  22. 22.
    Bruggeman BJ, Maier JA, Mohiuddin YS, Powers R et al (2012) Avian intervertebral disc arises from rostral sclerotome and lacks a nucleus pulposus: implications for evolution of the vertebrate disc. Dev Dyn 241(4):675–683PubMedPubMedCentralGoogle Scholar
  23. 23.
    Beckett MC, Ralphs JR, Caterson B, Hayes AJ (2015) The transmembrane heparan sulphate proteoglycan syndecan-4 is involved in establishment of the lamellar structure of the annulus fibrosus of the intervertebral disc. Eur Cell Mater 30:69–88 (discussion 88) PubMedGoogle Scholar
  24. 24.
    Takahashi Y, Yasuhiko Y, Takahashi J, Takada S et al (2013) Metameric pattern of intervertebral disc/vertebral body is generated independently of Mesp2/Ripply-mediated rostro-caudal patterning of somites in the mouse embryo. Dev Biol 380(2):172–184PubMedGoogle Scholar
  25. 25.
    Adams MA, Dolan P (2012) Intervertebral disc degeneration: evidence for two distinct phenotypes. J Anat 221(6):497–506PubMedPubMedCentralGoogle Scholar
  26. 26.
    Yin R, Lord EL, Cohen JR, Buser Z et al (2015) Distribution of Schmorl nodes in the lumbar spine and their relationship with lumbar disk degeneration and range of motion. Spine 40(1):E49–E53PubMedGoogle Scholar
  27. 27.
    Teraguchi M, Yoshimura N, Hashizume H, Muraki S et al (2015) The association of combination of disc degeneration, end plate signal change, and Schmorl node with low back pain in a large population study: the Wakayama Spine Study. Spine J 15(4):622–628PubMedGoogle Scholar
  28. 28.
    Walcott BP, Nahed BV, Mohyeldin A, Coumans JV, Kahle KT, Ferreira MJ (2012) Chordoma: current concepts, management, and future directions. Lancet Oncol 13(2):e69–e76PubMedGoogle Scholar
  29. 29.
    Choi KS, Harfe BD (2011) Hedgehog signaling is required for formation of the notochord sheath and patterning of nuclei pulposi within the intervertebral discs. Proc Natl Acad Sci U S A 108(23):9484–9489PubMedPubMedCentralGoogle Scholar
  30. 30.
    Stafford DA, Brunet LJ, Khokha MK, Economides AN, Harland RM (2011) Cooperative activity of noggin and gremlin 1 in axial skeleton development. Development 138(5):1005–1014PubMedPubMedCentralGoogle Scholar
  31. 31.
    Stafford DA, Monica SD, Harland RM (2014) Follistatin interacts with Noggin in the development of the axial skeleton. Mech Dev 131:78–85PubMedPubMedCentralGoogle Scholar
  32. 32.
    Ellis K, Bagwell J, Bagnat M (2013) Notochord vacuoles are lysosome-related organelles that function in axis and spine morphogenesis. J Cell Biol 200(5):667–679PubMedPubMedCentralGoogle Scholar
  33. 33.
    Buisson N, Sirour C, Moreau N, Denker E et al (2014) An adhesome comprising laminin, dystroglycan and myosin IIA is required during notochord development in Xenopus laevis. Development 141(23):4569–4579PubMedGoogle Scholar
  34. 34.
    McMillen P, Holley SA (2015) The tissue mechanics of vertebrate body elongation and segmentation. Curr Opin Genet Dev 32:106–111PubMedPubMedCentralGoogle Scholar
  35. 35.
    Gray RS, Wilm TP, Smith J, Bagnat M et al (2014) Loss of col8a1a function during zebrafish embryogenesis results in congenital vertebral malformations. Dev Biol 386(1):72–85PubMedGoogle Scholar
  36. 36.
    Wilting J, Kurz H, Brand-Saberi B, Steding G et al (1994) Kinetics and differentiation of somite cells forming the vertebral column: studies on human and chick embryos. Anat Embryol (Berl) 190(6):573–581Google Scholar
  37. 37.
    Li D, Yang H, Huang Y, Wu Y, Sun T, Li X (2014) Lumbar intervertebral disc puncture under C-arm fluoroscopy: a new rat model of lumbar intervertebral disc degeneration. Exp Anim 63(2):227–234PubMedPubMedCentralGoogle Scholar
  38. 38.
    Sakai D, Nishimura K, Tanaka M, Nakajima D et al (2015) Migration of bone marrow-derived cells for endogenous repair in a new tail-looping disc degeneration model in the mouse: a pilot study. Spine J 15(6):1356–1365PubMedGoogle Scholar
  39. 39.
    Kim KW, Kim YS, Ha KY, Woo YK et al (2009) An autocrine or paracrine Fas-mediated counterattack: a potential mechanism for apoptosis of notochordal cells in intact rat nucleus pulposus. Spine 30(11):1247–1251Google Scholar
  40. 40.
    Kim KW, Ha KY, Lee JS, Nam SW et al (2009) Notochordal cells stimulate migration of cartilage end plate chondrocytes of the intervertebral disc in in vitro cell migration assays. Spine J 9(4):323–329PubMedGoogle Scholar
  41. 41.
    Kim KW, Lim TH, Kim JG, Jeong ST, Masuda K, An HS (2003) The origin of chondrocytes in the nucleus pulposus and histologic findings associated with the transition of a notochordal nucleus pulposus to a fibrocartilaginous nucleus pulposus in intact rabbit intervertebral discs. Spine 28(10):982–990PubMedGoogle Scholar
  42. 42.
    Hunter CJ, Matyas JR, Duncan NA (2004) Cytomorphology of notochordal and chondrocytic cells from the nucleus pulposus: a species comparison. J Anat 205(5):357–362PubMedPubMedCentralGoogle Scholar
  43. 43.
    Thompson K, Moore S, Tang S, Wiet M, Purmessur D (2018) The chondrodystrophic dog: a clinically relevant intermediate-sized animal model for the study of intervertebral disc-associated spinal pain. JOR Spine 1(1):e1011PubMedPubMedCentralGoogle Scholar
  44. 44.
    Cai F, Wu XT, Xie XH, Wang F et al (2015) Evaluation of intervertebral disc regeneration with implantation of bone marrow mesenchymal stem cells (BMSCs) using quantitative T2 mapping: a study in rabbits. Int Orthop 39(1):149–159PubMedGoogle Scholar
  45. 45.
    Samartzis D, Borthakur A, Belfer I, Bow C et al (2015) Novel diagnostic and prognostic methods for disc degeneration and low back pain. Spine J 15(9):1919–1932PubMedPubMedCentralGoogle Scholar
  46. 46.
    Peng B, Hou S, Wu W, Zhang C, Yang Y (2006) The pathogenesis and clinical significance of a high-intensity zone (HIZ) of lumbar intervertebral disc on MR imaging in the patient with discogenic low back pain. Eur Spine J 15(5):583–587PubMedGoogle Scholar
  47. 47.
    Shi R, Wang F, Hong X, Wang YT et al (2015) The presence of stem cells in potential stem cell niches of the intervertebral disc region: an in vitro study on rats. Eur Spine J 24(11):2411–2424PubMedGoogle Scholar
  48. 48.
    Henriksson H, Thornemo M, Karlsson C, Hägg O et al (2009) Identification of cell proliferation zones, progenitor cells and a potential stem cell niche in the intervertebral disc region: a study in four species. Spine 34(21):2278–2287PubMedGoogle Scholar
  49. 49.
    Li Z, Peroglio M, Alini M, Grad S (2015) Potential and limitations of intervertebral disc endogenous repair. Curr Stem Cell Res Ther 10(4):329–338PubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Feng Wang
    • 1
    • 2
  • Cong Zhang
    • 1
    • 2
  • Arjun Sinkemani
    • 1
    • 2
  • Rui Shi
    • 1
    • 2
  • Zhi-Yang Xie
    • 1
    • 2
  • Lu Chen
    • 1
    • 2
  • Lu Mao
    • 1
    • 2
  • Xiao-Tao Wu
    • 1
    • 2
    Email author
  1. 1.Department of Spine Surgery, Zhongda Hospital, School of MedicineSoutheast UniversityNanjingChina
  2. 2.Surgery Research Center, School of MedicineSoutheast UniversityNanjingChina

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