Bioreactor cultivation of three-dimensional cartilage-carrier-constructs

A flow-chamber bioreactor was designed for generation of three-dimensional cartilage-carrier-constructs. A specific attribute of the flow-chamber is a very thin medium layer for improved oxygen supply and a counter current flow of medium and gas. Three-dimensional cartilage-carrier-constructs were produced according to a standard protocol from chondrocytes of an adult mini-pig. The final step of this protocol was performed either in the bioreactor or in 12-well plates. The bioreactor experiments showed a significantly higher matrix thickness but a lower ratio of glycosaminoglycan to DNA. For both culture methods the constructs contained a high amount of collagen II. Appearance of the cartilage obtained in the bioreactor seemed to be closer to native cartilage with respect to distribution of the cells within the matrix, smoothness of the surface etc. All results considered the flow-chamber bioreactor is a very useful tool for generation of three dimensional cartilage-carrier constructs.

[1]  W W Minuth,et al.  Artificial Tissues in Perfusion Culture , 1997, The International journal of artificial organs.

[2]  J. Petersen,et al.  Present and Future Therapies of Articular Cartilage Defects , 2003, European Journal of Trauma.

[3]  S. Waldman,et al.  Characterization of cartilagenous tissue formed on calcium polyphosphate substrates in vitro. , 2002, Journal of biomedical materials research.

[4]  H. Unno,et al.  Effect of shear stress on activity of cellular enzyme in animal cell , 1994 .

[5]  J. Vacanti,et al.  Tissue engineering : Frontiers in biotechnology , 1993 .

[6]  Ralf Pörtner,et al.  Perfusion cultures and modelling of oxygen uptake with three-dimensional chondrocyte pellets , 1999 .

[7]  J. Tramper,et al.  Oxygen gradients in tissue‐engineered Pegt/Pbt cartilaginous constructs: Measurement and modeling , 2004, Biotechnology and bioengineering.

[8]  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.

[9]  T. W. James,et al.  Assay for nanogram quantities of DNA in cellular homogenates. , 1979, Analytical biochemistry.

[10]  J. Jansen,et al.  Trabecular bone response to injectable calcium phosphate (Ca-P) cement. , 2002, Journal of biomedical materials research.

[11]  G. Vunjak‐Novakovic,et al.  Frontiers in Tissue Engineering , 1999 .

[12]  R. Dermietzel,et al.  Construction of an apparatus for perfusion cell cultures which enables in vitro experiments under organotypic conditions. , 1992, European journal of cell biology.

[13]  M J Lysaght,et al.  Demographic Scope and Economic Magnitude of Contemporary Organ Replacement Therapies , 2000, ASAIO journal.

[14]  J. Jansen,et al.  Histological evaluation of the bone response to calcium phosphate cement implanted in cortical bone. , 2003, Biomaterials.

[15]  Clemens A van Blitterswijk,et al.  Cartilage Tissue Engineering: Controversy in the Effect of Oxygen , 2003, Critical reviews in biotechnology.

[16]  J. McPherson,et al.  Synergistic action of transforming growth factor-beta and insulin-like growth factor-I induces expression of type II collagen and aggrecan genes in adult human articular chondrocytes. , 1997, Experimental cell research.

[17]  M. Morlock,et al.  Relationship between physical, biochemical and biomechanical properties of tissue-engineered cartilage-carrier-constructs , 2005, Biotechnology Letters.

[18]  M. J. Ward,et al.  Tissue-Engineered Cartilage for Implantation and Grafting , 1995, Facial plastic surgery : FPS.

[19]  Kyriacos A Athanasiou,et al.  Articular cartilage bioreactors and bioprocesses. , 2003, Tissue engineering.

[20]  M. Morlock,et al.  Calcium Phosphate Ceramics as Substrate for Cartilage Cultivation , 2004 .

[21]  J. Hassenpflug,et al.  The influence of oxygen and hydrostatic pressure on articular chondrocytes and adherent bone marrow cells in vitro. , 2004, Biorheology.

[22]  G. Vunjak‐Novakovic,et al.  Frontiers in tissue engineering. In vitro modulation of chondrogenesis. , 1999, Clinical orthopaedics and related research.