Comprehensive Histochemical Evaluation of Age-Related Intervertebral Disc Degeneration

Burcu Gültekin , Nihal Canbulat, Sena Nur Çelik , ERDAL KALKAN, Sabiha Serpil Kalkan

  • Burcu Gültekin: Necmettin Erbakan Üniversitesi Tıp Fakültesi
  • Nihal Canbulat: Necmettin Erbakan Üniversitesi Tıp Fakültesi
  • Sena Nur Çelik: Necmettin Erbakan Üniversitesi Tıp Fakültesi
  • ERDAL KALKAN: Konya Medova Hastanesi, Beyin ve Sinir Cerrahisi
  • Sabiha Serpil Kalkan: Necmettin Erbakan Üniversitesi Tıp Fakültesi
  • Year : 2026
  • Vol : 42
  • Issue : 1
  •  Page : 63-70
Objective: The intervertebral disc (IVD) is a fibrocartilaginous structure that plays a critical role in transmitting spinal loads and maintaining mobility. Age-related
degeneration of the IVD leads to the disruption of structural integrity, reduction in mechanical function, and the development of clinical symptoms such as back
and neck pain. During the degenerative process, matrix composition, cellular organization, and mechanical resilience are significantly affected. The aim of this study
was to investigate the histological and histochemical changes occurring in IVDs across different age groups using various staining techniques and to evaluate their
relationship with the degeneration process.
Materials and Methods: This study included 30 patients aged between 40 and 70 years who underwent surgical procedures for disc herniation during which IVD
specimens were obtained. Patients were divided into four age groups: Age40s (40–49 years, n=9), Age50s (50–59 years, n=7), Age60s (60–69 years, n=8), and Age70s
(70–79 years, n=6). Surgical specimens were fixed in 10% formalin, processed routinely, and embedded in paraffin. Sections of 5 μm in thickness were cut from
the paraffin blocks and stained with hematoxylin and eosin, toluidine blue, Masson’s trichrome, Congo red, and the Armed Forces Institute of Pathology method.
Histological changes were examined under a light microscope and semi-quantitatively scored.
Results: With advancing age, IVD tissues exhibited a marked decrease in proteoglycan content, an increase in collagen fiber density, accumulation of lipofuscin
granules, and the presence of amyloid deposits. In the older age groups, the matrix was observed to become denser and more fibrotic, and to show disruption of the
lamellar organization.
Conclusion: Histochemical staining techniques are effective in the detailed identification of cellular and extracellular matrix changes occurring during IVD aging
and degeneration. These methods contribute to the characterization of age-related structural alterations and provide valuable information for comparative
histopathological assessments of the degenerative process.
Cite this Article As : Gultekin B, Canbulat N, Celik SN, Kalkan E, Kalkan SS. Comprehensive Histochemical Evaluation of Age-Related Intervertebral Disc Degeneration. Selcuk Med J 2026;42(1): 63-70

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Conflict of interest : The authors declare that they have no conflict of interest.
Selcuk Medical Journal
2026, Vol. 42 (1)
ISSN: 1017-6616
E-ISSN: 2149-8059
Received : , Accepted : , Published Online :

References

  1. 1) Roughley PJ. Biology of intervertebral disc aging and degeneration: involvement of the extracellular matrix. Spine. 2004; 29:2691–99 doi: 10.1097/01.brs.0000146101.53784.b1.
  2. 2) Lai A, Gansau J, Gullbrand SE, et al. Development of a standardized histopathology scoring system for intervertebral disc degeneration in rat models: An initiative of the ORS spine section. JOR Spine. 2021; 26; 4(2): e1150. doi: 10.1002/jsp2.1150.
  3. 3) Postacchini F, Bellocci M, Massobrio M. Morphologic changes in annulus fibrosus during aging. An ultrastructural study in rats. Spine (Phila Pa 1976). 1984; 9(6): 596–03. doi:10.1097/00007632-198409000-00010.
  4. 4) Hayes AJ, Benjamin M, Ralphs JR. Role of actin stress fibres in the development of the intervertebral disc: cytoskeletal control of extracellular matrix assembly. Dev Dyn. 1999; 215(3):179–89. doi:10.1002/(SICI)1097-0177.
  5. 5) Bruehlmann SB, Rattner JB, Matyas JR, et al. Regional variations in the cellular matrix of the annulus fibrosus of the intervertebral disc. J Anat. 2002;201(2):159–71. doi:10.1046/j.1469-7580.2002.00080.x.
  6. 6) Wu Q, Huang JH. Intervertebral Disc Aging, Degeneration, and Associated Potential Molecular Mechanisms. J Head Neck Spine Surg. 2017;1(4):555569. doi: 10.19080/JHNSS.2017.01.555569.
  7. 7) Peletti-Figueiró M, Moura Aguiar I, Paesi S, et al. Histological markers of degeneration and regeneration of the human intervertebral disk. Coluna/Columna. 2017; 16 (1): 42–47. doi: 10.1590/s1808-185120171601170833.
  8. 8) Goldring MB, Goldring SR. Osteoarthritis. J Cell Physiol. 2007; 213: 626–34. doi: 10.1002/jcp.21258.
  9. 9) Cesari M, Kritchevsky SB, Leeuwenburgh C, et al. Oxidative damage and platelet activation as new predictors of mobility disability and mortality in elders. Antioxidants & redox signaling. 2006; 8(3-4): 609–19. doi: 10.1089/ars.2006.8.609.
  10. 10) Kumaresan S, Yoganandan N, Pintar FA, et al. Morphology of young and old cervical spine intervertebral disc tissues. Biomed Sci Instrum. 2000; 36: 141–46. doi: 10.1115/imece1999-0464.
  11. 11) Vo NV, Hartman RA, Patil PR, et al. Molecular mechanisms of biological aging in intervertebral discs. J Orthop Res. 2016; 34(8) :1289-06. doi: 10.1002/jor.23195.
  12. 12) Wininger AE, Phelps BM, Le JT, et al. Musculoskeletal pathology as an early warning sign of systemic amyloidosis: a systematic review of amyloid deposition and orthopedic surgery. BMC Musculoskelet Disord. 2021; 8, 22 (1): 51. doi: 10.1186/s12891-020-03912-z.
  13. 13) Athanasou NA, Kokubun S, West L, et al. Glycosaminoglycans in intervertebral disc amyloid deposits. Eur Spine J. 1995; 4(5): 308-12. doi: 10.1007/BF00301041.
  14. 14) Shu CC, Smith MM, Smith SM, et al. 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(5):1049. doi: 10.3390/ijms18051049.
  15. 15) Walter BA, Torre OM, Laudier D, et al. Form and function of the intervertebral disc in health and disease: a morphological and stain comparison study. J Anat. 2015; 227(6):707-16. doi: 10.1111/joa.12258.
  16. 16) Elmounedi N, Bahloul W, Aoui M, et al. Original animal model of lumbar disc degeneration. Libyan Journal of Medicine. 2023; 18(1). doi: 10.1080/19932820.2023.2212481.
  17. 17) Luna LG. Manual of histologic staining methods of the Armed Forces Institute of Pathology. 3rd Edition, McGraw-Hill, New York, 1968: 258.
  18. 18) Kalkan E, Akhan G, Kalkan S, et al. İnsan İntervertebral Disk Materyalinde Lipofussin Pigmenti. Med J SDU. 2009; 3(1).
  19. 19) Athanasou NA, Kokubun S, West L, et al. Glycosaminoglycans in intervertebral disc amyloid deposits. Eur Spine J. 1995; 4, 308–12. doi: 10.1007/BF00301041.
  20. 20) Miyamoto S, Yonenobu K, Ono K. Experimental cervical spondylosis in the mouse. Spine
  21. 1991; 16: 495–00. doi: 10.1097/00007632-199110001-00008.
  22. 21) Johnson WE, Eisenstein SM, Roberts S. Cell cluster formation in degenerate lumbar intervertebral discs is associated with increased disc cell proliferation. Connect. Tissue Res. 2021; 42: 197–07. doi: 10.3109/03008200109005650.
  23. 22) Zhao CQ, Jiang LS, Dai LY. Programmed cell death in intervertebral disc degeneration. Apoptosis. 2006; 11: 2079–88. doi: 10.1007/s10495-006-0290-7.
  24. 23) Saluja G, Fitzpatrick K, Bruce M, et al. Schmorl’s nodes (intervertebral herniations of intervertebral disc tissue) in two historic British populations. J Anat. 1986; 145: 87–96.
  25. 24) Gower WE, Pedrini V. Age-Related Variations in Protein‑Polysaccharides from Human Nucleus Pulposus, Annulus Fibrosus, and Costal Cartilage. Journal of Bone & Joint Surgery. 1969; 51(6): 1154–62.
  26. 25) Veroutis D, Kouroumalis A, Lagopati N, et al. Evaluation of senescent cells in intervertebral discs by lipofuscin staining. Mech Ageing Dev. 2021; 199:111564. doi: 10.1016/j.mad.2021.111564.
  27. 26) Meisel HJ, Ganey T, Hutton WC, et al. Clinical experience in cell-based therapeutics: intervention and outcome. Eur Spine J. 2006; 15 (3): 397-05. doi: 10.1007/s00586-006-0169-x.
  28. 27) Gruber HE, Ingram JA, Norton HJ, et al. Senescence in cells of the aging and degenerating intervertebral disc: immunolocalization of senescence-associated beta-galactosidase in human and sand rat disc. Spine. 2007; 32(3): 321–27. doi: 10.1097/01.brs.0000253960.57051.de.
  29. 28) Freemont AJ, Watkins A, Le Maitre C, et al. Nerve growth factor expression and innervation of the painful intervertebral disc. J Pathol. 2002; 197: 286–92. doi: 10.1002/path.1108.
  30. 29) Johnson WE, Evans H, Menage J, et al. Immunohistochemical detection of Schwann cells in innervated and vascularized human intervertebral discs. Spine. 2001; 26: 2550–2557. doi: 10.1097/00007632-200112010-00007.
  31. 30) Kauppila LI. Ingrowth of blood vessels in disc degeneration. Angiographic and histological studies of cadaveric spines. J Bone Joint Surg Am. 1995; 77(1): 26‐31. doi: 10.2106/00004623-199501000-00004.
  32. 31) Johnson WEB, Patterson AM, Eisenstein SM, et al. The presence of pleiotrophin in the human intervertebral disc is associated with increased vascularization: an immunohistologic study. Spine. 2007; 32(12): 1295‐02. doi: 10.1097/BRS.0b013e31805b835d.
  33. 32) Stefanakis M, Al‐Abbasi M, Harding I, et al. Annulus fissures are mechanically and chemically conducive to the ingrowth of nerves and blood vessels. Spine. 2012; 37(22): 1883‐91. doi: 10.1097/BRS.0b013e318263ba59.
  34. 33) Urban JP, Roberts S. Degeneration of the intervertebral disc. Arthritis Res Ther. 2003; 5 (3): 120-30. doi: 10.1186/ar629.
  35. 34) Dimozi A, Mavrogonatou E, Sklirou A, et al. Oxidative stress inhibits the proliferation, induces premature senescence and promotes a catabolic phenotype in human nucleus pulposus intervertebral disc cells. Eur Cell Mater. 2015;30: 89-102. doi: 10.22203/ecm.v030a07.
  36. 35) Zhao L, Tian B, Xu Q, et al. Extensive mechanical tension promotes annulus fibrosus cell senescence through suppressing cellular autophagy. Biosci Rep. 2019; 39 (4). doi: 10.1042/BSR20190163.
  37. 36) Wullbrand A, Saeger W, Missmahl HP, et al. Amyloid in intervertebral discs of surgery and autopsy material. A new class of amyloid? Virchows Archiv. A, Pathological anatomy and histopathology. 1990; 416(4): 335–41. doi: 10.1007/BF01605294.
  38. 37) Madhani A, Kotturu N, Fine D, et al. Spinal amyloid deposits are common among older patients undergoing spinal stenosis decompression surgery. J Gen Fam Med. 2025; 15, 26(3): 222-30. doi: 10.1002/jgf2.766.
  39. 38) Antoniou J, Steffen T, Nelson F, et al. The human lumbar intervertebral disc: Evidence for changes in the biosynthesis and denaturation of the extracellular matrix with growth, maturation, ageing, and degeneration. Journal of Clinical Investigation. 1996; 98(4): 996-03. doi: 10.1172/JCI118884.
  40. 39) Boos N, Weissbach S, Rohrbach H, et al. Classification of age-related changes in lumbar intervertebral discs. Spine. 2002; 27: 2631–44. doi: 10.1097/00007632-200212010-00002.