Hydrostatic pressure modulates proteoglycan metabolism in chondrocytes seeded in agarose.

OBJECTIVE To investigate the effect of isolated hydrostatic pressure on proteoglycan metabolism in chondrocytes. METHODS Bovine articular chondrocytes cultured in agarose gels were subjected to 5 MPa hydrostatic pressure for 4 hours in either a static or a pulsatile (1 Hz) mode, and changes in glycosaminoglycan (GAG) synthesis, hydrodynamic size, and aggregation properties of proteoglycans and aggrecan messenger RNA (mRNA) levels were determined. RESULTS The application of 5 MPa static pressure caused a significant increase in GAG synthesis of 11% (P < 0.05). Column chromatography showed that this increase in GAG synthesis was associated with large proteoglycans. In addition, semiquantitative reverse transcriptase-polymerase chain reaction showed a 4-fold increase in levels of aggrecan mRNA (P < 0.01). CONCLUSION Hydrostatic pressure in isolation, which does not cause cell deformation, can affect proteoglycan metabolism in chondrocytes cultured in agarose gels, indicating an important role of hydrostatic pressure in the regulation of extracellular matrix turnover in articular cartilage.

[1]  C. Handley,et al.  Modulation of aggrecan and link-protein synthesis in articular cartilage. , 1992, The Biochemical journal.

[2]  J. Urban,et al.  The effects of hydrostatic pressure on matrix synthesis in articular cartilage , 1991, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[3]  S. Kumar,et al.  A simplified in situ solubilization procedure for the determination of DNA and cell number in tissue cultured mammalian cells. , 1985, Analytical biochemistry.

[4]  T. Sandström,et al.  The effect of continuous mechanical pressure upon the turnover of articular cartilage proteoglycans in vitro. , 1982, Clinical orthopaedics and related research.

[5]  P. Ragan,et al.  Down‐regulation of chondrocyte aggrecan and type‐II Collagen gene expression correlates with increases in static compression magnitude and duration , 1999, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[6]  R. Schneiderman,et al.  Effects of mechanical and osmotic pressure on the rate of glycosaminoglycan synthesis in the human adult femoral head cartilage: An in vitro study , 1986, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[7]  D. R. Carter,et al.  In vitro stimulation of articular chondrocyte mRNA and extracellular matrix synthesis by hydrostatic pressure , 1996, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[8]  K. Brandt,et al.  Effects of static and cyclic compressive loading on articular cartilage plugs in vitro. , 1984, Arthritis and rheumatism.

[9]  P. Benya,et al.  Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels , 1982, Cell.

[10]  A. Grodzinsky,et al.  Fluorometric assay of DNA in cartilage explants using Hoechst 33258. , 1988, Analytical biochemistry.

[11]  W H Simon,et al.  Scale effects in animal joints. I. Articular cartilage thickness and compressive stress. , 1970, Arthritis and rheumatism.

[12]  W. A. Hodge,et al.  Contact pressures in the human hip joint measured in vivo. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[13]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[14]  H. Helminen,et al.  Effects of cyclic hydrostatic pressure on proteoglycan synthesis in cultured chondrocytes and articular cartilage explants. , 1993, Archives of biochemistry and biophysics.

[15]  D L Bader,et al.  Compressive strains at physiological frequencies influence the metabolism of chondrocytes seeded in agarose , 1997, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[16]  S W O'Driscoll,et al.  Periosteum responds to dynamic fluid pressure by proliferating in vitro , 1999, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[17]  H. Mankin,et al.  In vitro metabolic response of articular cartilage segments to low levels of hydrostatic pressure. , 1985, Connective tissue research.

[18]  A. Grodzinsky,et al.  Kinetics of the chondrocyte biosynthetic response to compressive load and release. , 1989, Biochimica et biophysica acta.

[19]  V. Mow,et al.  Load-controlled compression of articular cartilage induces a transient stimulation of aggrecan gene expression. , 1998, Archives of biochemistry and biophysics.

[20]  A. Grodzinsky,et al.  Biosynthetic response of cartilage explants to dynamic compression , 1989, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[21]  G. Lust,et al.  Characterization of cartilage metabolic response to static and dynamic stress using a mechanical explant test system. , 1997, Journal of biomechanics.

[22]  H. Helminen,et al.  Influence of short‐term hydrostatic pressure on organization of stress fibers in cultured chondrocytes , 1995, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[23]  J. Bryan Finlay,et al.  Instrumentation and Procedure for the Controlled Impact of Articular Cartilage , 1978, IEEE Transactions on Biomedical Engineering.

[24]  R M Aspden,et al.  Effects of mechanical load on cartilage matrix biosynthesis in vitro. , 1991, Matrix.

[25]  van de Stadt Rj,et al.  Effects of loading on the synthesis of proteoglycans in different layers of anatomically intact articular cartilage in vitro. , 1992, The Journal of rheumatology.

[26]  T. Hardingham,et al.  The effects of cycloheximide on the biosynthesis and secretion of proteoglycans by chondrocytes in culture. , 1981, The Biochemical journal.

[27]  E. Thonar,et al.  Quantification of 35S-labeled proteoglycans complexed to alcian blue by rapid filtration in multiwell plates. , 1994, Analytical biochemistry.

[28]  T D Brown,et al.  Techniques for mechanical stimulation of cells in vitro: a review. , 2000, Journal of biomechanics.

[29]  Takashi Nakamura,et al.  Establishment of bone morphogenetic protein 2 responsive chondrogenic cell line , 1996, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[30]  W C Hutton,et al.  Contact pressures in the human hip joint. , 1987, The Journal of bone and joint surgery. British volume.

[31]  A. Grodzinsky,et al.  Compression of cartilage results in differential effects on biosynthetic pathways for aggrecan, link protein, and hyaluronan. , 1996, Archives of biochemistry and biophysics.

[32]  P. Roughley,et al.  Changes in messenger RNA and protein levels of proteoglycans and link protein in human osteoarthritic cartilage samples. , 1997, Arthritis and rheumatism.

[33]  G. Nuki,et al.  Hyperpolarisation of cultured human chondrocytes following cyclical pressure‐induced strain: Evidence of a role for α5β1 integrin as a chondrocyte mechanoreceptor , 1997 .

[34]  A. Grodzinsky,et al.  Cartilage electromechanics--I. Electrokinetic transduction and the effects of electrolyte pH and ionic strength. , 1987, Journal of biomechanics.

[35]  M B Aydelotte,et al.  Differences between sub-populations of cultured bovine articular chondrocytes. II. Proteoglycan metabolism. , 1988, Connective tissue research.

[36]  D R Carter,et al.  Time-dependent effects of intermittent hydrostatic pressure on articular chondrocyte type II collagen and aggrecan mRNA expression. , 2000, Journal of rehabilitation research and development.

[37]  Macdonald Ag.  Physiological Aspects of Deep Sea Biology , 1975 .

[38]  A. Grodzinsky,et al.  Chondrocytes in agarose culture synthesize a mechanically functional extracellular matrix , 1992, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.