Abstract
There is evidence that a fall in nutrient supply leads to disc degeneration but little understanding of the effects of nutrient deprivation on the physiology of disc cells which govern the composition of the disc. We examined the effects of changes in glucose and oxygen concentration and pH on the viability and metabolism of cells from bovine nucleus pulposus. Cells isolated from bovine discs and embedded in alginate beads were cultured under oxygen and glucose concentrations from zero to physiological levels and maintained at pH 7.4, pH 6.7, or pH 6.2 for up to 3 days. Interactions between nutrient concentrations were examined in relation to cell viability and lactic acid production. Cell viability was significantly reduced in the absence of glucose, with or without oxygen. Disc cells survived at 0% oxygen, provided that glucose was present, as seen previously. Cell viability decreased if the medium was acidic, more so when combined with low glucose concentrations. The rate of lactic acid production also fell as the pH became acidic and after 24 h or more at low glucose concentrations, but it did not appear to vary with oxygen concentration under the culture conditions used here. Glucose, rather than oxygen, appears to be the nutrient critical for maintaining disc cell viability. However, in an avascular tissue such as the disc, it is unlikely that glucose deprivation will occur alone; it will almost certainly correlate with a fall in oxygen concentration and pH. These results indicate that the combined nutrient and metabolite environment, rather than concentrations of any single nutrient, should be considered when studying cellular physiology in the disc.
Similar content being viewed by others
References
Aoki J, Yamamoto I, Kitamura N, Sone T, Itoh H, Torizuka K, Takasu K (1987) End plate of the discovertebral joint: degenerative change in the elderly adult. Radiology 164: 411–414
Baker MS, Feigan J, Lowther DA (1989) The mechanism of chondrocyte hydrogen peroxide damage. Depletion of intracellular ATP due to suppression of glycolysis caused by oxidation of glyceraldehyde-3-phosphate dehydrogenase. J Rheumatol 16: 7–14
Bartels EM, Fairbank JCT, Winlove CP, Urban JPG (1998) Oxygen and lactate concentrations measured in vivo in the intervertebral discs of scoliotic and back pain patients. Spine 23: 1–8
Battie MC, Videman T, Gill K, Moneta GB, Nyman R, Kaprio J, Koskenvuo M (1991) 1991 Volvo Award in clinical sciences. Smoking and lumbar intervertebral disc degeneration: an MRI study of identical twins. Spine 16: 1015–1021
Bibby SR (2002) Cell metabolism and viability in the intervertebral disc. Dr Phil thesis, Oxford University
Brodin H (1955) Paths of nutrition in articular cartilage and intervertebral discs. Acta Orthop Scand 24: 177–183
Brown MD, Tsaltas T (1976) Studies on the permeability of the intervertebral disc during skeletal maturation. Spine 1: 240–244
Casciari JJ, Sotirchos SV, Sutherland RM (1992) Mathematical modeling of microenvironment and growth in emt6/ro multicellular tumor spheroids. Cell Proliferation 25: 1–22
Chelberg MK, Banks GM, Geiger DF, Oegema TR Jr (1995) Identification of heterogeneous cell populations in normal human intervertebral disc. J Anat 186 (Pt 1): 43–53
Diamant B, Karlsson J, Nachemson A (1968) Correlation between lactate levels and pH in discs of patients with lumbar rhizopathies. Experientia 24: 1195–1196
Errington RJ, Puustjarvi K, White IR, Roberts S, Urban JP (1998) Characterisation of cytoplasm-filled processes in cells of the intervertebral disc. J Anat 192 (Pt 3): 369–378
Esumi H, Izuishi K, Kato K, Hashimoto K, Kurashima Y, Kishimoto A, Ogura T, Ozawa T (2002) Hypoxia and nitric oxide treatment confer tolerance to glucose starvation in a 5’-AMP-activated protein kinase-dependent manner. J Biol Chem 277: 32791–32798
Grimshaw MJ, Mason RM (2000) Bovine articular chondrocyte function in vitro depends upon oxygen tension. Osteoarthritis Cartilage 8: 386–392
Grimshaw MJ, Mason RM (2001) Modulation of bovine articular chondrocyte gene expression in vitro by oxygen tension. Osteoarthritis Cartilage 9: 357–364
Hansen U, Schunke M, Domm C, Ioannidis N, Hassenpflug J, Gehrke T, Kurz B (2001) Combination of reduced oxygen tension and intermittent hydrostatic pressure: a useful tool in articular cartilage tissue engineering. J Biomech 34: 941–949
Hassler O (1970) The human intervertebral disc. A micro-angiographical study of its vascular supply at various ages. Acta Orthop Scand 40: 765–772
Holm S, Nachemson A (1983) Cellularity in the intervertebral disc and its relevance to nutrition. Proceedings of the International Society for the Study of the Lumbar Spine
Holm S, Maroudas A, Urban JP, Selstam G, Nachemson A (1981) Nutrition of the intervertebral disc: solute transport and metabolism. Connect Tissue Res 8: 101–119
Horner HA, Urban JP (2001) 2001 Volvo Award winner in basic science studies: effect of nutrient supply on the viability of cells from the nucleus pulposus of the intervertebral disc. Spine 26: 2543–2549
Horner HA, Roberts S, Bielby RC, Menage J, Evans H, Urban J P (2002) Cells from different regions of the intervertebral disc: effect of culture system on matrix expression and cell phenotype. Spine 27: 1018–1028
Ishihara H, Urban JP (1999) Effects of low oxygen concentrations and metabolic inhibitors on proteoglycan and protein synthesis rates in the intervertebral disc. J Orthop Res 17: 829–835
Ishihara H, Warensjo K, Roberts S, Urban JP (1997) Proteoglycan synthesis in the intervertebral disk nucleus: the role of extracellular osmolality. Am J Physiol 272: C1499-C1506
Kauppila LI, Penttila A, Karhunen PJ, Lalu K, Hannikainen P (1994) Lumbar disc degeneration and atherosclerosis of the abdominal aorta. Spine 19: 923–929
Lee RB, Urban JP (1997) Evidence for a negative Pasteur effect in articular cartilage. Biochem J 321: 95–102
Lee RB, Urban JP (2002) Functional replacement of oxygen by other oxidants in articular cartilage. Arthritis Rheum 46: 3190–3200
Lee RB, Wilkins RJ, Razaq S, Urban JP (2002) The effect of mechanical stress on cartilage energy metabolism. Biorheology 39: 133–143
Maldonado BA, Oegema-TR J (1992) Initial characterization of the metabolism of intervertebral disc cells encapsulated in microspheres. J Orthop Res 10: 677–690
McFadden KD, Taylor JR (1989) End-plate lesions of the lumbar spine. Spine 14: 867–869
Nachemson A, Lewin T, Maroudas A, Freeman MAF (1970) In vitro diffusion of dye through the end-plates and annulus fibrosus of human lumbar intervertebral discs. Acta Orthop Scand 41: 589–607
Ohshima H, Urban JPG (1992) Effect of lactate concentrations and pH on matrix synthesis rates in the intervertebral disc. Spine 17: 1079–1082
Razaq S, Wilkins RJ, Urban JPG (2003) The effect of extracellular pH on matrix turnover by cells of the bovine nucleus pulposus. Europ Spine J 12: 341–349
Roberts S, Menage J, Eisenstein SM (1993) The cartilage end-plate and intervertebral disc in scoliosis: calcification and other sequelae. J Orthop Res 11: 747–757
Roberts S, Urban JPG, Evans H, Eisenstein SM (1996) Transport properties of the human cartilage endplate in relation to its composition and calcification. Spine 21: 415–420
Selard E, Shirazi-Adl AS, Urban JPG (2003) Computational studies of nutrient transport in human intervertebral disc. Spine (in press)
Stairmand J, Holm S, Urban J (1991) Factors influencing oxygen concentration gradients in the intervertebral disc: a theoretical analysis. Spine 16: 444–449
Urban JP, Holm S, Maroudas A, Nachemson A (1977) Nutrition of the intervertebral disk. An in vivo study of solute transport. Clin Orthop 101–114
Urban MR, Fairbank JC, Etherington PJ, Loh FL, Winlove CP, Urban JP (2001) Electrochemical measurement of transport into scoliotic intervertebral discs in vivo using nitrous oxide as a tracer. Spine 26: 984–990
Ysart GE, Mason RM (1994) Responses of articular cartilage expiant cultures to different oxygen tensions. Biochim Biophys Acta 1221: 15–20
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Bibby, S.R.S., Urban, J.P.G. Effect of nutrient deprivation on the viability of intervertebral disc cells. Eur Spine J 13, 695–701 (2004). https://doi.org/10.1007/s00586-003-0616-x
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s00586-003-0616-x