Skip to main content
Erschienen in: European Spine Journal 3/2006

01.08.2006 | Original Article

Regulation of gene expression in intervertebral disc cells by low and high hydrostatic pressure

verfasst von: Cornelia Neidlinger-Wilke, Karin Würtz, Jill P. G. Urban, Wolfgang Börm, Markus Arand, Anita Ignatius, Hans-Joachim Wilke, Lutz E. Claes

Erschienen in: European Spine Journal | Sonderheft 3/2006

Einloggen, um Zugang zu erhalten

Abstract

Intervertebral disc structures are exposed to wide ranges of intradiscal hydrostatic pressure during different loading excercises and are at their minimum during lying or relaxed sitting and at maximum during lifting weights with a round back. We hypothesize that these different loading magnitudes influence the intervertebral disc (IVD) by alteration of disc matrix turnover depending on their magnitudes. Therefore the aim of this study was to assess changes in gene expression of human nucleus cells after the application of low hydrostatic pressure (0.25 MPa) and high hydrostatic pressure (2.5 MPa). IVD cells isolated from the nucleus of human (n = 18) and bovine (n = 24 from four animals) disc biopsies were seeded into three-dimensional collagen type-I matrices and exposed to the different loading magnitudes by specially developed pressure chambers. The lower pressure range (0.25 MPa, 30 min, 0.1 Hz) was applied with a recently published device by using an external compression cylinder. For the application of higher loads (2.5 MPa, 30 min, 0.1 Hz) the cell-loaded collagen gels were sealed into sterile bags with culture medium and stimulated in a newly developed water-filled compression cylinder by using a loading frame. These methods allowed the comparison of loading regimes in a wide physiological range under an equal three-dimensional culture conditions. Cells were harvested 24 h after the end of stimulation and changes in the expression of genes known to influence IVD matrix turnover (collagen-I, collagen-II, aggrecan, MMP1, MMP2, MMP3, MMP13) were analyzed by real-time RT-PCR. A Wilcoxon signed-rank test1 and a Wilcoxon 2-sample test2 were performed to detect differences between the stimulated and control samples1 and differences between low and high hydrostatic pressure2. Multiple testing was considered by adjusting the p value appropriately. Both regimes of hydrostatic pressure influenced gene expression in nucleus cells with opposite tendencies for the matrix forming proteins aggrecan and collagen type-I in response to the two different pressure magnitudes: Low hydrostatic-pressure (0.25 MPa) tended to increase collagen-I and aggrecan expression of human nucleus cells (P < 0.05) but only to a small degree. High hydrostatic pressure (2.5 MPa) tended to decrease gene expression of all anabolic proteins with significant effects on aggrecan expression of nucleus cells (P = 0.004). Low hydrostatic pressure had no influence on the expression of matrix metalloproteinases (MMP1, MMP2, MMP3 and MMP13). In contrast, high hydrostatic pressure tended to increase the expression of MMP1, MMP3 and MMP13 of human nucleus cells with high individual-individual variations. The decreased expression of aggrecan (P = 0.008) and collagen type II (P = 0.023) and the increased MMP3 expression (P = 0.008) in response to high hydrostatic pressure could be confirmed in additional experiments with bovine nucleus cells. These results suggest that hydrostatic pressure as one of the physiological stimuli of the IVD may influence matrix turnover in a magnitude dependent way. Low hydrostatic pressure (0.25 MPa) has quite small influences with a tendency to anabolic effects, whereas high hydrostatic pressure (2.5 MPa) tends to decrease the matrix protein expression with a tendency to increase some matrix-turnover enzymes. Therefore, hydrostatic pressure may regulate disc matrix turnover in a dose-dependent way.
Literatur
1.
Zurück zum Zitat Adams MA, McNally DS, Dolan P (1996) ‘Stress’ distributions inside intervertebral discs. The effects of age and degeneration. J Bone Joint Surg Br 78(6):965–72PubMedCrossRef Adams MA, McNally DS, Dolan P (1996) ‘Stress’ distributions inside intervertebral discs. The effects of age and degeneration. J Bone Joint Surg Br 78(6):965–72PubMedCrossRef
2.
Zurück zum Zitat Bibby SR, Urban JP (2004) Effect of nutrient deprivation on the viability of intervertebral disc cells. Eur Spine J 13 (8):695–701PubMedCrossRef Bibby SR, Urban JP (2004) Effect of nutrient deprivation on the viability of intervertebral disc cells. Eur Spine J 13 (8):695–701PubMedCrossRef
3.
Zurück zum Zitat Brand RA, Stanford CM, Nicolella DP (2001) Primary adult human bone cells do not respond to tissue (continuum) level strains. J Orthop Sci 6(3):295–301PubMedCrossRef Brand RA, Stanford CM, Nicolella DP (2001) Primary adult human bone cells do not respond to tissue (continuum) level strains. J Orthop Sci 6(3):295–301PubMedCrossRef
4.
Zurück zum Zitat Buckwalter JA (1995) Aging and degeneration of the human intervertebral disc. Spine 20(11):1307–14PubMed Buckwalter JA (1995) Aging and degeneration of the human intervertebral disc. Spine 20(11):1307–14PubMed
5.
Zurück zum Zitat Fehrenbacher A, Steck E, Rickert M, Roth W, Richter W (2003) Rapid regulation of collagen but not metalloproteinase 1, 3, 13, 14 and tissue inhibitor of metalloproteinase 1, 2, 3 expression in response to mechanical loading of cartilage explants in vitro. Arch Biochem Biophys 410(1):39–47PubMedCrossRef Fehrenbacher A, Steck E, Rickert M, Roth W, Richter W (2003) Rapid regulation of collagen but not metalloproteinase 1, 3, 13, 14 and tissue inhibitor of metalloproteinase 1, 2, 3 expression in response to mechanical loading of cartilage explants in vitro. Arch Biochem Biophys 410(1):39–47PubMedCrossRef
6.
Zurück zum Zitat Frost HM (1990) Skeletal structural adaptations to mechanical usage (SATMU): 4. Mechanical influences on intact fibrous tissues. Anat Rec 226(4):433–439PubMedCrossRef Frost HM (1990) Skeletal structural adaptations to mechanical usage (SATMU): 4. Mechanical influences on intact fibrous tissues. Anat Rec 226(4):433–439PubMedCrossRef
7.
Zurück zum Zitat Hall AC (1999) Differential effects of hydrostatic pressure on caution transport pathways of isolated articular chondrocytes. J Cell Physiol 178(2):197–204PubMedCrossRef Hall AC (1999) Differential effects of hydrostatic pressure on caution transport pathways of isolated articular chondrocytes. J Cell Physiol 178(2):197–204PubMedCrossRef
8.
Zurück zum Zitat Handa T, Ishihara H, Ohshima H, Osada R, Tsuji H, Obata K (1997) Effects of hydrostatic pressure on matrix synthesis and matrix metalloproteinase production in the human lumbar intervertebral disc. Spine 22(10):1085–1091PubMedCrossRef Handa T, Ishihara H, Ohshima H, Osada R, Tsuji H, Obata K (1997) Effects of hydrostatic pressure on matrix synthesis and matrix metalloproteinase production in the human lumbar intervertebral disc. Spine 22(10):1085–1091PubMedCrossRef
9.
Zurück zum Zitat Ishihara H, McNally DS, Urban JP, Hall AC (1996) Effects of hydrostatic pressure on matrix synthesis in different regions of the intervertebral disk. J Appl Physiol 80(3):839–846PubMed Ishihara H, McNally DS, Urban JP, Hall AC (1996) Effects of hydrostatic pressure on matrix synthesis in different regions of the intervertebral disk. J Appl Physiol 80(3):839–846PubMed
10.
Zurück zum Zitat Kasra M, Goel V, Martin J, Wang ST, Choi W, Buckwalter J (2003) Effect of dynamic hydrostatic pressure on rabbit intervertebral disc cells. J Orthop Res 21(4):597–603PubMedCrossRef Kasra M, Goel V, Martin J, Wang ST, Choi W, Buckwalter J (2003) Effect of dynamic hydrostatic pressure on rabbit intervertebral disc cells. J Orthop Res 21(4):597–603PubMedCrossRef
11.
Zurück zum Zitat Maclean JJ, Lee CR, Alini M, Iatridis JC (2004) Anabolic and catabolic mRNA levels of the intervertebral disc vary with the magnitude and frequency of in vivo dynamic compression. J Orthop Res 22(6):1193–1200PubMedCrossRef Maclean JJ, Lee CR, Alini M, Iatridis JC (2004) Anabolic and catabolic mRNA levels of the intervertebral disc vary with the magnitude and frequency of in vivo dynamic compression. J Orthop Res 22(6):1193–1200PubMedCrossRef
12.
Zurück zum Zitat Neidlinger-Wilke C, Wurtz K, Liedert A et al (2005) A three-dimensional collagen matrix as a suitable culture system for the comparison of cyclic strain and hydrostatic pressure effects on intervertebral disc cells. J Neurosurg Spine 2(4):457–465PubMedCrossRef Neidlinger-Wilke C, Wurtz K, Liedert A et al (2005) A three-dimensional collagen matrix as a suitable culture system for the comparison of cyclic strain and hydrostatic pressure effects on intervertebral disc cells. J Neurosurg Spine 2(4):457–465PubMedCrossRef
13.
Zurück zum Zitat Setton LA, Chen J (2004) Cell mechanics and mechanobiology in the intervertebral disc. Spine 29(23):2710–2723PubMedCrossRef Setton LA, Chen J (2004) Cell mechanics and mechanobiology in the intervertebral disc. Spine 29(23):2710–2723PubMedCrossRef
14.
Zurück zum Zitat Shirazi-Adl A, Ahmed AM, Shrivastava SC (1986) A finite element study of a lumbar motion segment subjected to pure sagittal plane moments. J Biomech 19(4):331–350PubMedCrossRef Shirazi-Adl A, Ahmed AM, Shrivastava SC (1986) A finite element study of a lumbar motion segment subjected to pure sagittal plane moments. J Biomech 19(4):331–350PubMedCrossRef
15.
Zurück zum Zitat Stanford CM, Morcuende JA, Brand RA (1995) Proliferative and phenotypic responses of bone-like cells to mechanical deformation. J Orthop Res 13(5):664–670PubMedCrossRef Stanford CM, Morcuende JA, Brand RA (1995) Proliferative and phenotypic responses of bone-like cells to mechanical deformation. J Orthop Res 13(5):664–670PubMedCrossRef
16.
Zurück zum Zitat Stokes IA, Iatridis JC (2004) Mechanical conditions that accelerate intervertebral disc degeneration: overload versus immobilization. Spine 29(23):2724–2732PubMedCrossRef Stokes IA, Iatridis JC (2004) Mechanical conditions that accelerate intervertebral disc degeneration: overload versus immobilization. Spine 29(23):2724–2732PubMedCrossRef
17.
Zurück zum Zitat Urban JP, Roberts S (2003) Degeneration of the intervertebral disc. Arthritis Res Ther 5(3):120–130PubMedCrossRef Urban JP, Roberts S (2003) Degeneration of the intervertebral disc. Arthritis Res Ther 5(3):120–130PubMedCrossRef
18.
Zurück zum Zitat Videman T, Leppavuori J, Kaprio J et al (1998) Intragenic polymorphisms of the vitamin D receptor gene associated with intervertebral disc degeneration. Spine 23(23):2477–2485PubMedCrossRef Videman T, Leppavuori J, Kaprio J et al (1998) Intragenic polymorphisms of the vitamin D receptor gene associated with intervertebral disc degeneration. Spine 23(23):2477–2485PubMedCrossRef
19.
Zurück zum Zitat Videman T, Gibbons LE, Battie MC et al (2001) The relative roles of intragenic polymorphisms of the vitamin d receptor gene in lumbar spine degeneration and bone density. Spine 26(3):E7–E12PubMedCrossRef Videman T, Gibbons LE, Battie MC et al (2001) The relative roles of intragenic polymorphisms of the vitamin d receptor gene in lumbar spine degeneration and bone density. Spine 26(3):E7–E12PubMedCrossRef
20.
Zurück zum Zitat Wilke HJ, Neef P, Caimi M, Hoogland T, Claes LE (1999) New in vivo measurements of pressures in the intervertebral disc in daily life. Spine 24(8):755–762PubMedCrossRef Wilke HJ, Neef P, Caimi M, Hoogland T, Claes LE (1999) New in vivo measurements of pressures in the intervertebral disc in daily life. Spine 24(8):755–762PubMedCrossRef
Metadaten
Titel
Regulation of gene expression in intervertebral disc cells by low and high hydrostatic pressure
verfasst von
Cornelia Neidlinger-Wilke
Karin Würtz
Jill P. G. Urban
Wolfgang Börm
Markus Arand
Anita Ignatius
Hans-Joachim Wilke
Lutz E. Claes
Publikationsdatum
01.08.2006
Verlag
Springer-Verlag
Erschienen in
European Spine Journal / Ausgabe Sonderheft 3/2006
Print ISSN: 0940-6719
Elektronische ISSN: 1432-0932
DOI
https://doi.org/10.1007/s00586-006-0112-1

Weitere Artikel der Sonderheft 3/2006

European Spine Journal 3/2006 Zur Ausgabe

Arthropedia

Grundlagenwissen der Arthroskopie und Gelenkchirurgie. Erweitert durch Fallbeispiele, Videos und Abbildungen. 
» Jetzt entdecken

Update Orthopädie und Unfallchirurgie

Bestellen Sie unseren Fach-Newsletter und bleiben Sie gut informiert.