Semi-continuous perfusion system for delivering intermittent physiological pressure to regenerating cartilage.

A semi-continuous compression/perfusion system has been custom made to allow the application of intermittent hydrostatic pressure, at physiological levels, to regenerating tissues over the long term. To test the system, isolated foal chondrocytes were seeded in resorbable polyglycolic acid meshes and cultured in the system for 5 weeks. The cell/polymer constructs were subjected to an intermittent hydrostatic pressure of 500 psi and were fed semi-continuously. Assays of the resulting tissue constructs indicate that the reactor supports cartilage development and that physiological intermittent compression enhances the production of extracellular matrix by the chondrocytes. The concentrations of sulfated glycosaminoglycan were found to be at least twice as high as those in control (unpressurized) samples. A correlation between the sulfated glycosaminoglycan content and the compressive modulus in pressurized, but not control, samples suggests that physiological intermittent pressurization not only enhances the production of extracellular matrix but may also influence matrix organization resulting in a stronger construct.

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

[2]  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.

[3]  G. Vunjak‐Novakovic,et al.  Cultivation of cell‐polymer cartilage implants in bioreactors , 1993, Journal of cellular biochemistry.

[4]  A. Grodzinsky,et al.  Effects of compression on the loss of newly synthesized proteoglycans and proteins from cartilage explants. , 1991, Archives of biochemistry and biophysics.

[5]  J. Veldhuijzen,et al.  Cartilage response to mechanical force in high-density chondrocyte cultures. , 1985, Arthritis and rheumatism.

[6]  M. Klagsbrun.  Large-scale preparation of chondrocytes. , 1979, Methods in enzymology.

[7]  Gordana Vunjak-Novakovic,et al.  Microgravity tissue engineering , 1997, In Vitro Cellular & Developmental Biology - Animal.

[8]  C. Brighton,et al.  The effect of oxygen tension on proteoglycan synthesis and aggregation in mammalian growth plate chondrocytes , 1991, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

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

[10]  D. Buttle,et al.  Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. , 1986, Biochimica et biophysica acta.

[11]  G. Lust,et al.  Effect of compressive loading and unloading on the synthesis of total protein, proteoglycan, and fibronectin by canine cartilage explants , 1993, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[12]  Gordana Vunjak-Novakovic,et al.  Effects of mixing on the composition and morphology of tissue‐engineered cartilage , 1996 .

[13]  G. Vunjak‐Novakovic,et al.  Mass transfer studies of tissue engineered cartilage. , 1996, Tissue engineering.

[14]  S. Lohmander.  Turnover of proteoglycans in guinea pig costal cartilage. , 1977, Archives of biochemistry and biophysics.

[15]  J. F. Woessner,et al.  The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. , 1961, Archives of biochemistry and biophysics.

[16]  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.

[17]  R. Todhunter,et al.  Effects of exercise and polysulfated glycosaminoglycan on repair of articular cartilage defects in the equine carpus , 1993, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[18]  C. Malemud,et al.  Biomechanical properties of third carpal articular cartilage in exercised and nonexercised horses , 1995, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[19]  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.

[20]  A. Dorfman,et al.  THE GROWTH OF CARTILAGE CELLS IN SOFT AGAR AND LIQUID SUSPENSION , 1970, The Journal of cell biology.

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

[22]  B. Prahl-Andersen,et al.  The growth of cartilage cells in vitro and the effect of intermittent compressive force. A histological evaluation. , 1987, Connective tissue research.

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

[24]  Shannon R. Magari,et al.  Mini‐review: Mechanical factors affecting cartilage regeneration in vitro , 2000, Biotechnology and bioengineering.

[25]  W. D. Morgan,et al.  In vitro culture of articular chondrocytes , 1973 .

[26]  L. Peterson,et al.  The repair of experimentally produced defects in rabbit articular cartilage by autologous chondrocyte transplantation , 1989, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[27]  G. Vunjak‐Novakovic,et al.  Composition of cell‐polymer cartilage implants , 1994, Biotechnology and bioengineering.

[28]  S L Woo,et al.  Dynamic behavior of a biphasic cartilage model under cyclic compressive loading. , 1995, Journal of biomechanics.

[29]  R. Salter,et al.  The biological effect of continuous passive motion on the healing of full-thickness defects in articular cartilage. An experimental investigation in the rabbit. , 1980, The Journal of bone and joint surgery. American volume.