Advertisement

Biomedical Microdevices

, Volume 13, Issue 4, pp 789–798 | Cite as

Development of high-throughput perfusion-based microbioreactor platform capable of providing tunable dynamic tensile loading to cells and its application for the study of bovine articular chondrocytes

  • Min-Hsien Wu
  • Hsin-Yao Wang
  • Heng-Liang Liu
  • Shih-Siou Wang
  • Yen-Ting Liu
  • Yan-Ming Chen
  • Shiao-Wen Tsai
  • Chun-Li Lin
Article

Abstract

Mammalian cells are sensitive to extracellular microenvironments. In order to precisely explore the physiological responses of cells to tensile loading, a stable and well-defined culture condition is required. In this study, a high-throughput perfusion-based microbioreactor platform capable of providing dynamic equibiaxial tensile loading to the cultured cells under a steady culture condition was proposed. The mechanism of generating tensile stimulation to cells is based on the pneumatically-driven deformation of an elastic polydimethylsiloxan (PDMS) membrane which exerts tensile loading to the attached cells. By modulating the magnitude and frequency of the applied pneumatic pressure, various tensile loading can be generated in a controllable manner. In this study, the microbioreactor platform was designed with the aid of the experimentally-validated finite element (FE) analysis to ensure the loading of tensile strain to cells is uniform and definable. Based on this design, the quantitative relationship between the applied pneumatic pressure and the generated tensile strain on the PDMS membrane was established via FE analysis. Results demonstrated that the proposed device was able to generate the tensile strain range (0~0.12), which covers the physiological condition that articular chondrocytes experience tensile strain under human walking condition. In this study, moreover, the effect of tensile loading on the metabolic, biosynthetic and proliferation activities of articular chondrocytes was investigated. Results disclosed that the dynamic tensile loading of 0.12 strain at 1 Hz might significantly up-regulate the synthesis of glycosaminoglycans while such stimulation was found no significant influence on the metabolic activity, the synthesis of collagen, and the proliferation of chondrocytes. Overall, this study has presented a high throughput perfusion micro cell culture device that is suitable for precisely exploring the effect of tensile loading on cell physiology.

Keywords

Microbioreactors Tensile strain Mechanical stimulations Perfusion culture High throughput Articular chondrocytes 

Notes

Acknowledgments

The authors would like to thank financial support from the National Science Council in Taiwan (97-2218-E-182-002 MY2) and Chang Gung Memorial Hospital (CMRPD170281 and CMRPD170282).

References

  1. K. Boxshall, M.H. Wu, Z. Cui, Z.F. Cui, J.F. Watts, M.A. Baker, Simple surface treatments to modify protein adsorption and cell attachment properties within a Poly(dimethylsiloxane) microbioreactor. Surf. Interface Anal. 38, 198–201 (2006)CrossRefGoogle Scholar
  2. J.A. Buckwalter, L.C. Rosenberg, E.B. Hunziker, Articular cartilage: composition, structure and response to injury, and methods of facilitating repair, in Articular cartilage and knee joint function: basic science and arthroscopy, ed. by J.W. Ewing (Raven, New York, 1990), pp. 19–56Google Scholar
  3. D.T. Eddington, W.C. Crone, D.J. Beebe, Development of Process Protocols to Fine Tune Polydimethylsiloxane Material Properties. (7th International Conference on Miniaturized Chemical and Biochemical Analysis Systems, 2003), pp. 1089–1092Google Scholar
  4. J.C. Fan, S.D. Waldman, The effect of intermittent static biaxial tensile strains on tissue engineered cartilage. Ann. Biomed. Eng. 38, 1672–1682 (2010)CrossRefGoogle Scholar
  5. R.W. Farndale, D.J. Buttle, A.J. Barrett, Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim. Biophys. Acta. 883, 173–177 (1986)Google Scholar
  6. A.M. Freyria, M.C. Ronziere, S. Roche, C.F. Rousseau, D. Herbage, Regulation of growth, protein synthesis and maturation of foetal bovine chondrocytes grown in high-density culture in the presence of ascorbic acid, retinoic acid and dihydrocytochalasin B. J. Cell. Biochem. 76, 84–98 (1999)CrossRefGoogle Scholar
  7. O. Gabay, D.J. Hall, F. Berenbaum, Y. Henrotin, C. Canchez, Osteoarthritis and obesity: experimental models. Joint Bone Spine 75, 675–679 (2008)CrossRefGoogle Scholar
  8. T.M. Griffin, F. Guilak, The role of mechanical loading in the onset and progression of osteoarthritis. Exerc. Sport Sci. Rev. 33, 195–200 (2005)CrossRefGoogle Scholar
  9. Y. Hirano, N. Ishiguro, M. Sokabe, M. Takigawa, K. Naruse, Effects of tensile and compressive strains on response of a chondrocytic cell line embedded in type I collagen gel. J. Biotechnol. 133, 245–252 (2008)CrossRefGoogle Scholar
  10. C.D. Hoemann, J. Sun, V. Chrzanowski, M.D. Buschmann, A multivalent assay to detect glycosaminoglycan, protein, collagen, RNA, DNA content in milligram samples of cartilage or hydrogel- based repair cartilage. Anal. Biochem. 300, 1–10 (2002)CrossRefGoogle Scholar
  11. K. Honda, S. Ohno, K. Tanimoto, C. Ijuin, N. Tanaka, T. Doi, Y. Kato, K. Tanne, The effects of high magnitude cyclic tensile load on cartilage matrix metabolism in cultured chondrocytes. Eur. J. Cell Biol. 79, 601–609 (2000)CrossRefGoogle Scholar
  12. J. Huang, L.R. Ballou, K.A. Hasty, Cyclic equibiaxial tensile strain induces both anabolic and catabolic responses in articular chondrocytes. Gene 404, 101–109 (2007)CrossRefGoogle Scholar
  13. P. Julkunen, W. Wilson, J.S. Jurvelin, R.K. Korhonen, Composition of the pericellular matrix modulates the deformation behavior of chondrocytes in articular cartilage under static loading. Med. Biol. Eng. Comput. 47, 1281–1290 (2009)CrossRefGoogle Scholar
  14. G. Knutsen, L. Engebretsen, T.C. Ludvigsen, J.O. Drogset, T. Grontvedt, E. Solheim, T. Strand, S. Roberts, V. Isaksen, O. Johansen, Autologous chondrocyte implantation compared with microfracture in the knee: a randomized trial. J. Bone Joint Surg. Am. 86, 455–464 (2004)Google Scholar
  15. C.Y. Lee, H.C. Hsu, X. Zhang, D.Y. Wang, Z.P. Luo, Cyclic compression and tension regulate differently the metabolism of chondrocytes. J. Musculosketeletal. Res. 9, 59–64 (2005)MATHCrossRefGoogle Scholar
  16. C.L. Lin, J.C. Wang, Y.C. Kuo, Numerical simulation on the biomechanical interactions of tooth/implant-supported system under various occlusal forces with rigid/non-rigid connections. J. Biomech. 39, 453–463 (2006)CrossRefGoogle Scholar
  17. J.L. Lin, M.H. Wu, C.Y. Kuo, K.D. Lee, Y.L. Shen, Application of indium tin oxide (ITO)-based microheater chip with uniform thermal distribution for perfusion cell culture outside a cell incubator. Biomed. Microdevices 12, 389–398 (2010)CrossRefGoogle Scholar
  18. Y.H. Lin, S.W. Kang, T.Y. Wu, Fabrication of polydimethylsiloxane (PDMS) pulsating heat pipe. Appl. Therm. Eng. 29, 573–580 (2009)CrossRefGoogle Scholar
  19. T. Mawatari, D.P. Lindsey, A.H. Harris, S.B. Goodman, W.J. Maloney, R.L. Smith, Effects of tensile strain and fluid flow on osteoarthritic human chondrocyte metabolism in vitro. J. Orthop. Res. 28, 907–913 (2010)Google Scholar
  20. C. Moraes, J.H. Chen, Y. Sun, C.A. Simmons, Microfabricated arrays for high-throughput screening of cellular response to cyclic substrate deformation. Lab Chip 10, 227–234 (2010)CrossRefGoogle Scholar
  21. R.M. Schulz, A. Bader, Cartilage tissue engineering and bioreactor systems for the cultivation and stimulation of chondrocytes. Eur. Biophys. J. 36, 539–568 (2007)CrossRefGoogle Scholar
  22. H. Stegemann, K.H. Stalder, Determination of hydroxyproline. Clin. Chim. Acta 18, 267–273 (1967)CrossRefGoogle Scholar
  23. A.V. Spuskanyuk, R.M. McMeeking, V.S. Deshpande, E. Arzt, The effect of shape on the adhesion of fibrillar surfaces. Acta. Biomater. 4, 1669–1676 (2008)CrossRefGoogle Scholar
  24. M. Ueki, N. Tanaka, K. Tanimoto, C. Nishio, K. Honda, Y.Y. Lin, Y. Tanne, S. Ohkuma, T. Kamiya, E. Tanaka, K. Tanne, The effect of mechanical loading on the metabolism of growth plate chondrocytes. Ann. Biomed. Eng. 36, 793–800 (2008)CrossRefGoogle Scholar
  25. J.P. Urban, J.F. McMullin, Swelling pressure of the intervertebral disc: influence of proteoglycan and collagen content. Biorheology 22, 145–157 (1985)Google Scholar
  26. E.J. Vanderploeg, C.G. Wilson, M.E. Levenston, Articular chondrocytes derived from distinct tissue zones differentially respond to in vitro oscillatory tensile loading. Osteoarthr Cartil 16, 1228–1236 (2008)CrossRefGoogle Scholar
  27. J.L. Williams, J.H. Chen, D.M. Belloli, Strain fields on cell stressing devices employing clamped circular elastic diaphragms as substrates. J. Biomech. Eng. 114, 377–384 (1992)CrossRefGoogle Scholar
  28. M.H. Wu, S.B. Huang, Z. Cui, Z. Cui, G.B. Lee, Development of perfusion-based micro 3-D cell culture platform and its application for high throughput drug testing. Sens. Actuators, B, Chem. 129, 231–240 (2008)CrossRefGoogle Scholar
  29. M.H. Wu, J.P. Urban, Z. Cui, Z.F. Cui, Development of PDMS microbioreactor with well-defined and homogenous culture environment for chondrocyte 3-D culture. Biomed. Microdevices 8, 331–340 (2006)CrossRefGoogle Scholar
  30. M.H. Wu, J.P.G. Urban, Z.F. Cui, Z. Cui, X. Xu, Effect of extracellular pH on matrix synthesis by chondrocytes in 3D agarose gel. Biotechnol. Prog. 23, 430–434 (2007)CrossRefGoogle Scholar
  31. Z. Xu, M.J. Buckley, C.H. Evans, S. Agarwal, Cyclic tensile strain acts as an antagonist of IL-1 beta actions in chondrocytes. J. Immunol. 165, 453–460 (2000)Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Min-Hsien Wu
    • 1
  • Hsin-Yao Wang
    • 2
  • Heng-Liang Liu
    • 3
  • Shih-Siou Wang
    • 1
  • Yen-Ting Liu
    • 1
  • Yan-Ming Chen
    • 1
  • Shiao-Wen Tsai
    • 1
  • Chun-Li Lin
    • 4
  1. 1.Graduate Institute of Biochemical and Biomedical EngineeringChang Gung UniversityTaoyuanTaiwan
  2. 2.School of MedicineChang Gung UniversityTaoyuanTaiwan
  3. 3.Department of Mechanical EngineeringChang Gung UniversityTaoyuanTaiwan
  4. 4.Department of Biomedical EngineeringNational Yang-Ming UniversityTaipeiTaiwan

Personalised recommendations