The influence of serum, glucose and oxygen on intervertebral disc cell growth in vitro: implications for degenerative disc disease

Article English OPEN
Johnson, William EB ; Stephan, Simon ; Roberts, Sally (2008)
  • Publisher: BioMed Central
  • Journal: Arthritis Research & Therapy, volume 10, issue 2, pages R46-R46 (issn: 1478-6354, eissn: 1478-6362)
  • Related identifiers: doi: 10.1186/ar2405, pmc: PMC2453766
  • Subject: Research Article | /dk/atira/pure/subjectarea/asjc/2700 | Rheumatology | Medicine(all) | Immunology and Allergy | /dk/atira/pure/subjectarea/asjc/2700/2745 | Immunology
    mesheuropmc: musculoskeletal system | musculoskeletal diseases

Introduction: The avascular nature of the human intervertebral disc (IVD) is thought to play a major role in disc pathophysiology by limiting nutrient supply to resident IVD cells. In the human IVD, the central IVD cells at maturity are normally chondrocytic in phenotype. However, abnormal cell phenotypes have been associated with degenerative disc diseases, including cell proliferation and cluster formation, cell death, stellate morphologies, and cell senescence. Therefore, we have examined the relative influence of possible blood-borne factors on the growth characteristics of IVD cells in vitro. Methods: Bovine IVD cells were cultured either in monolayer to encourage cell proliferation or in alginate to induce chondrocytic differentiation. In both culture systems, cells were maintained with or without 20% serum, with or without 320 mg/dL glucose, and in atmospheric levels (∼21%) of oxygen or 1% oxygen. Cell proliferation and viability, cell senescence, and collagen immunopositivity were assessed after 7 days. Statistical differences in these growth characteristics were tested using nonparametric analyses (n = 4 samples). Results: In both culture systems, serum deprivation significantly inhibited IVD cell proliferation and increased cell positivity for senescence-associated beta-galactosidase (SA-β-gal), a marker of cell senescence. Conversely, IVD cells cultured in the presence of serum, but deprived of glucose, proliferated significantly more rapidly. In alginate cultures, this enhanced cell proliferation (through glucose deprivation) led to the formation of IVD cell clusters. Serum-deprived cells in monolayer, but not in alginate, adopted a stellate appearance. Oxygen deprivation alone had little effect on IVD cell proliferation or survival. Oxygen and glucose deprivation also had no significant effect on SA-β-gal positivity. IVD cell viability was markedly and significantly decreased in serum-deprived alginate cultures, but in all other conditions remained at or greater than approximately 95%. Glucose deprivation, but not serum or oxygen deprivation, inhibited synthesis of type I and type II collagen, both in monolayer and alginate cultures. Conclusion: This study demonstrates that factors present in serum interact with other nutrients, notably glucose, to play a major role in regulating the behaviour of IVD cells. These findings suggest that IVD cell phenotypes seen in degenerative disc disease may arise through the cells' response to altered vascularisation and nutrient supply. © 2008 Johnson et al.; licensee BioMed Central Ltd.
  • References (38)
    38 references, page 1 of 4

    1. Maroudas A, Stockwell RA, Nachemson A, Urban J: Factors involved in the nutrition of the human lumbar intervertebral disc: cellularity and diffusion of glucose in vitro. J Anat 1975, 120:113-130.

    2. Gruber HE, Hanley EN Jr: Analysis of aging and degeneration of the human intervertebral disc. Comparison of surgical specimens with normal controls. Spine 1998, 23:751-757.

    3. 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 2002, 27:2631-2644.

    4. Johnson WE, 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.

    5. Johnson WE, Roberts S: Human intervertebral disc cell morphology and cytoskeletal composition: a preliminary study of regional variations in health and disease. J Anat 2003, 203:605-612.

    6. Roberts S, Evans EH, Kletsas D, Jaffray DC, Eisenstein SM: Senescence in human intervertebral discs. Eur Spine J 2006, 15:S312-316.

    7. Gruber HE, Ingram JA, Norton HJ, Hanley EN Jr: Senescence in cells of the aging and degenerating intervertebral disc: immunolocalization of senescence-associated beta-galactosidase in human and sand rat discs. Spine 2007, 32:321-327.

    8. Le Maitre CL, Freemont AJ, Hoyland JA: Accelerated cellular senescence in degenerate intervertebral discs: a possible role in the pathogenesis of intervertebral disc degeneration. Arthritis Res Ther 2007, 9:R45.

    9. Urban JP, Smith S, Fairbank JC: Nutrition of the intervertebral disc. Spine 2004, 29:2700-2709.

    10. Grunhagen T, Wilde G, Soukane DM, Shirazi-Adl SA, Urban JP: Nutrient supply and intervertebral disc metabolism. J Bone Joint Surg Am 2006, 88:30-35.

  • Metrics
    No metrics available
Share - Bookmark