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Bioprocess and Biosystems Engineering

, Volume 27, Issue 4, pp 273–280 | Cite as

Bioreactor cultivation of three-dimensional cartilage-carrier-constructs

  • Stephanie Nagel-Heyer
  • Christiane Goepfert
  • Frank Feyerabend
  • Jan Philipp Petersen
  • Peter Adamietz
  • Norbert M. Meenen
  • Ralf PörtnerEmail author
Original papers

Abstract

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.

Keywords

Bioreactor Cartilage tissue engineering Chondrocytes Flow-chamber Immunohistology 

Notes

Acknowledgements

The financial support of Biomet Deutschland GmbH, Darmstadt, Germany under the BMBF-grant No. 03N4012 is gratefully acknowledged. Furthermore we thank Katharina Braun, Ditte Siemesgelüss for their excellent technical support as well as Prof. Dr. Michael Morlock for his advice in statistical analysis.

References

  1. 1.
    Langer R, Vacanti JP (1993) Tissue engineering. Science 260:920–926Google Scholar
  2. 2.
    Lysaght MJ, Loughlin JA (2000) Demographic scope and economic magnitude of contemporary organ replacement therapies. ASAIO J 46:515Google Scholar
  3. 3.
    Minuth WW, Strehl R, Schumacher K (2005) Tissue engineering—from cell biology to artificial organs. Wiley-VCH, WeinheimGoogle Scholar
  4. 4.
    Freed LE, Martin I, Vunjak-Novakovic G (1999) Frontiers in tissue engineering. In: vitro modulation of chondrogenesis. Clin Orthop 367(Suppl):46–58Google Scholar
  5. 5.
    Buschmann MD, Gluzband YA, Grodzinsky AJ, Kimura JH, Hunziker EB (1992) Chondrocytes in agarose culture synthesize a mechanically functional extrazellular matrix. J Orthop Res 10:745–758CrossRefGoogle Scholar
  6. 6.
    Park SS, Ward MJ (1995) Tissue-engineered cartilage for implantation and grafting. Facial Plastic Surg 11:278–283Google Scholar
  7. 7.
    Sittinger M, Schultz O, Keyser G, Minuth WW, Burmester GR (1997) Artificial tissues in perfusion culture. Int J Artif Org 20:57–62Google Scholar
  8. 8.
    Petersen JP, Rücker A, von Stechow D, Adamietz P, Pörtner R, Rueger JM, Meenen NM (2003) Present and future therapies of articular cartilage defects. Eur J Trauma 1:1–10Google Scholar
  9. 9.
    Nagel-Heyer S, Goepfert Ch, Morlock MM, Pörtner R (2005) Relationship between gross morphological and biochemical data of tissue engineered cartilage-carrier-constructs. Biotechnol Lett 27:187–192CrossRefGoogle Scholar
  10. 10.
    Nehring D, Adamietz P, Meenen NM, Pörtner R (1999) Perfusion cultures and modelling of oxygen uptake with three-dimensional chondrocyte pellets. Biotechnol Techn 13:701–706CrossRefGoogle Scholar
  11. 11.
    Minuth WW, Stöckl G, Kloth S, Dermietzel R (1992) Construction of an apparatus for perfusion cell cultures which enables in vitro experiments under organotypic conditions. Eur J Cell Biol 57:132–137Google Scholar
  12. 12.
    Yaeger PC, Masi TL, Buck de Ortiz JL, Binette F, Tubo R, MyPherson JM (1997) Synergistic action of transforming growth factor-β and insulin-like growth factor-I induces expression of type II collagen and aggrecan genes in adult human articular chondrocytes. Exp Cell Res 237:318–325CrossRefGoogle Scholar
  13. 13.
    Janssen R, Nagel-Heyer S, Goepfert CH, Pörtner R, Toykan D, Krummhauer O, Morlock MM, Adamietz P, Meenen NM, Kriven WM, Kim D-K, Tampieri A, Celotti G (2004) Calcium phosphate ceramics as substrate for cartilage cultivation. Ceramic Eng Sci Proc 25(4):523–528Google Scholar
  14. 14.
    Scherer K, Schunke M, Sellckau R, Hassenpflug J, Kurz B (2004) The influence of oxygen and hydrostatic pressure on articular chondrocytes and adherent bone marrow cells in vitro. Biorheology 41:323–333Google Scholar
  15. 15.
    Brunk CF, Jones KC, James TW (1979) Assay for Nanogram Quantities of DNA in Cellular Homogenates. Anal Biochem 92:497–500CrossRefGoogle Scholar
  16. 16.
    Junqueira LC, Carneiro J (1991) Histologie. Springer, Berlin Heidelberg New YorkGoogle Scholar
  17. 17.
    Malda J, Martens DE, Tramper J, van Blitterswijk CA, Riesle J (2003) Cartilage tissue engineering: controversy in the effect of oxygen. Crit Rev Biotechnol 23:175–194Google Scholar
  18. 18.
    Malda J, Rouwkema J, Martens DE, Le Comte EP, Kooy FK, Tramper J, van Blitterswijk CA, Riesle J (2004) Oxygen gradients in tissue-engineered PEGT/PBT cartilaginous constructs: measurement and modeling. Biotechnol Bioeng 86:9–18CrossRefGoogle Scholar
  19. 19.
    Ooms EM, Wolke JGC, van der Waerden JPCM, Jansen JA (2002) Trabecular bone response to injectable calcium phosphate (Ca–P) cement. J Biomed Mater Res 61:9–18CrossRefGoogle Scholar
  20. 20.
    Ooms EM, Wolke JGC, van de Heuvel MT, Jeschke B, Jansen JA (2003) Histological evaluation of the bone response to calcium phosphate cement implanted in cortical bone. Biomaterial 24:989–1000Google Scholar
  21. 21.
    Darling EM, Athanasiou KA (2003) Articular cartilage bioprocesses and bioreactors. Tissue Eng 9:9–26Google Scholar
  22. 22.
    Minuth WW, Strehl R, Schumacher K (2005) Tissue engineering—from cell biology to artificial organs. Wiley-VCH, WeinheimGoogle Scholar
  23. 23.
    Shiragami N, Unno H (1994) Effect of shear stress on activity of cellular enzyme in animal cell. Bioproc Eng 10:43–45Google Scholar
  24. 24.
    Brandt KD, Doherty M, Lohmander LS (1998) Composition and structure of articular cartilage. In: Osteoarthritis. Oxford University Press, New York, pp 110–111Google Scholar
  25. 25.
    Waldman SD, Grynpas MD, Pilliar RM, Kandel RA (2002) Characterization of cartilaginous tissue formed on calcium polyphosphate substrate in vitro. J Biomed Mat Res 62:323–330Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Stephanie Nagel-Heyer
    • 1
  • Christiane Goepfert
    • 1
  • Frank Feyerabend
    • 2
  • Jan Philipp Petersen
    • 3
  • Peter Adamietz
    • 2
  • Norbert M. Meenen
    • 3
  • Ralf Pörtner
    • 1
    Email author
  1. 1.Bioprozess- und BioverfahrenstechnikTechnische Universität Hamburg-HarburgHamburgGermany
  2. 2.Institut für Biochemie und Molekularbiologie IIUniversitätsklinikum EppendorfHamburgGermany
  3. 3.Unfall-, Hand- und WiederherstellungschirurgieUniversitätsklinikum EppendorfHamburgGermany

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