Development of a Tensile Cell Stimulator to Study the Effects of Uniaxial Tensile Stress on Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells

세포 인장 자극기의 개발과 세포 인장 자극을 통한 성체 줄기세포의 골분화 유도

  • 신현준 (한국과학기술원 기계공학과) ;
  • 이우택 (한국과학기술원 기계공학과) ;
  • 박석훈 (한국과학기술원 기계공학과) ;
  • 이선화 (고려대학교 의과대학 정형외과) ;
  • 박정호 (고려대학교 의과대학 정형외과) ;
  • 윤용산 (한국과학기술원 기계공학과) ;
  • 신현정 (한국과학기술원 기계공학과)
  • Published : 2009.07.01


Mechanical stimulation is known to play a vital role on the differentiation of mesenchymal stem cells (MSCs) to pre-osteoblasts. In this research, we developed a tensile cell stimulator, composed of a DC motor-driven actuator and LVDT sensor for measuring linear displacement, to study the effects of tensile stress on osteogenic differentiation of MSCs. First, we demonstrated the reliability of this device by showing the uniform strain field in the silicon substrate. Secondly, we investigated the effects of tensile stretching on osteogenic differentiation. We imposed a pre-set cyclic strain at a fixed frequency on cell monolayer cultured on a flexible silicon substrate while varying its amplitude and duration. 60 min of resting period was allowed between 30 min of cyclic stretching and this cycle is repeated up to 7 days. Under the combined stimulation with osteogenic media and mechanical stretching, the osteogenic markers such as alkaline phosphatase (ALP), osterix, and osteopontin began to get expressed as early as 4 days of stimulation, which is much shorter than what is typically required for osteogenic media induced differentiation. Moreover, different markers were induced at different magnitudes of the applied strains. Lastly, for the case of ALP, we observed the antagonistic effects of osteogenic media when combined with mechanical stretching.


Osteogenic Differentiation;Mesenchymal Stem Cell;Strain Ratio;Cell Tensile Stimulation;2D Elastic Membrane;Quantitative Analysis;Osteogenic Marker


  1. Arnold, I. C., 2005, 'Mesenchymal Stem Cells: Cell-Based Reconstructive Therapy in Orthopedics,' Tissue Eng., Vol.11, No. 7/8, pp. 1198-1211
  2. Ignatius, A., Blessing, H., Liedert, A., Schmidt, C., Neidlinger-Wilke, C., Kaspar, D., Friemert B. and Claes, L., 2005, 'Tissue Engineering of Bone: Effects of Mechanical Strain on Osteoblastic Cells in Type I Collagen Matrices,' Biomaterials, Vol. 26, No. 3, pp.311-318
  3. Xiao, L., Marie, O., Wei, H. and Morris, K., 2005, 'Neural Cell Differentiation In Vitro from Adult Human Bone Marrow Mesenchymal Stem Cells,' Stem Cells Dev, Vol. 14, No. 1, pp.65-69
  4. Michelle, R., Kreke, M.E. and Aaron, S. G, 2004, 'Hydrodynamic Shear Stimulates Osteocalcin Expression But Not Proliferation of Bone Marrow Stromal Cells,' Tissue Eng, Vol. 10, No. 5/6, pp. 780-788
  5. Heng, B. C., Cao, T. and Lee, E. H., 2004, 'Directing Stem Cell Differentiation into the Chondrogenic Lineage In Vitro,' Stem Cells, Vol. 22, pp. 1152-1167
  6. Gruber, R., Graninger, W., Bobacz, K., Watzek, G., Erlacher, L, 2003, 'BMP-6-Induced Osteogenic Differentiation of Mesenchymal Cell Lines is not Modulated by Sex Steroids and Resveratrol,' Cytokine, Vol. 23, No. 4, pp. 133-137
  7. Jagodzinski, M., Drescher, M., Zeichen, J., Hankemeier, S., Krettek, C., Bosch, U. and Griensven, M., 2004, 'Effects Of Cyclic Longitudinal Mechanical Strain and Dexamethason on Osteogenic Differentiation of Human Bone,' Eur Cell Mater, Vol. 7, pp. 35-41
  8. Lee, E. K., Lee, J. S., Park, H. S., Kim, C. H, Gin, Y. J. and Son Y. S., 2005, 'Cyclic Stretch Stimulates Cell Proliferation of Human Mesenchymal Stem Cells but Do Not Induce Their Apoptosis and Differentiation,' Tissue Eng Reg Med, Vol. 2, No. 1, pp. 29-33
  9. Koike, M., Shimokawa, H., Keiichi K., Ohya, Z. K. and Kunimichi S., 2005, 'Effects of Mechanical Strain on Proliferation and Differentiation of Bone Marrow Stromal Cell Line ST2,' J Bone Miner Metab, Vol. 23, N0. 3, pp. 219-225
  10. Robling, A. G.,Castillo A. B., and Turner C. H., 2006, 'Biomechanical and Molecular Regulation of Bone Remodeling,' Annu Rev Biomed Eng, Vol.8, pp.455-498
  11. Altman, G., Horan, R., Martin, I., Farhadi, J., Srark, P., Volloch, V., Vunjak-novakovic, G., Richmond, J., Kaplan, D., L., 2002, 'Cell Differentiation by Mechanical Stress,' FASEB, Vol. 16, pp. 270-272
  12. Matsumoto, T., Delafontaine, P., Schnetzer, K.J., Tong, B.C., Nerem, R.M., 1996, 'Effect of Uniaxial, Cyclic Stretch on the Morphology of Monocytes/ Macrophages in Culture.' J Biomech Eng, Vol. 118, No. 3, pp.420-422
  13. Ives, C. L., Skin, S. G. and McIntire, L. V., 1986, 'Mechanical Effects on Endothelial Cell Morphology: in vitro Assessment.' In Vitro Cell Dev Biol, Vol. 22, No. 9, pp. 500-507
  14. Clark, C. B., Burkholder, T. J. and Frangos, J. A., 2001, 'Uniaxial Strain System to Investigate Strain Rate Regulation in Vitro.,' Rev Sci Instr, Vol. ;72, No. 5, pp. 2415-2422
  15. Matheson, L. A., Fairbank N. J., Maksym, G. N., J. Paul, Santerre and Labow R. S., 2006, 'Characterization of the $Flexcell^{TM} Uniflex^{TM}$ Cyclic Strain Culture System with U937 Macrophage-like Cells,' Biomatrial, Vol. 27, No. 2, pp. 226-233
  16. Burr, D.B., Milgrom, C., Fyhrie, D., Forwood, M., Nyska, M., Finestone, A., Hoshaw, S., Saiag, E., Simkin, A., 1996, 'In Vivo Measurement of Human Tibial Strains During Vigorous Activity,' Bone, Vol. 18, No. 5, pp.405-410
  17. Frank, O., Heim, M., Jakob, M., Barbero, A., Schafer, D., Bendik, I., Dick W., Heberer, M, Martin, I., 2002, 'Real-Time Quantitative RT-PCR Analysis of Human Bone Marrow Stromal Cells During Osteogenic Differentiation in Vitro,' J Cell Biochem., Vol. 85, pp. 737-746
  18. Bruder, S. P., Horowitz, M. C., Mosca, J. D., Hayneworth, S. E., 1997, 'Monoclonal Antibodies Reactive with Human Osteogenic Cell Surface Antigens,' Bone, Vol. 21, No. 3, pp.225-235
  19. Mauney, J. R., Sjostorm, S., Blumberg, J., Horan, R., O'Leary, J. P., Vunjak-Novakovic. G., Volloch, V. and Kaplan, D. L., 2004, 'Mechanical Stimulation Promotes Osteogenic Differentiation of Human Bone Marrow Stromal Cells on 3-D Partially Demineralized Bone Scaffolds In Vitro,' Calcif Tissue Int, Vol.74, No. 5, pp.458-468
  20. Byers, R.J., Brown, J., Brandwood, C., Wood, P., Staley, W., Hainey, L., Freemont, A.J. and Hoyland, J.A., 1999, 'Osteoblastic Differentiation and mRNA Analysis of STRO-1-Positive Human Bone Marrow Stromal Cells Using Primary in Vitro Culture and Poly (A) PCR.,' J Pathol, Vol. 187, No. 3, pp. 374-381<374::AID-PATH257>3.0.CO;2-V
  21. Leung, D.Y., Glagov, S. and Mathews, M.B., 1977, 'A New in vitro System for Studying Cell Response to Mechanical Stimulation. Different Effects of Cyclic Stretching and Agitation on Smooth Muscle Cell Biosynthesis,' Exp Cell Res, Vol. 109, No. 2, pp. 285-298
  22. Berry, C. C, Shelton, J. C., Bader, D.L. and Lee, D.A., 2003, 'Influence of External Uniaxial Cyclic Strain on Oriented Fibroblast-Seeded Collagen Gels.,' Tissue Eng, Vol. 9, No. 4, pp. 613-624
  23. Armbruster, C., Schneider, M., Gamerdinger, K., Schumann, S., Cuevas, M., Koch, E., Guttmann, J., 2008, 'Fabrication of thin and Flexible PDMS Membranes for Biomechanical Test Applications,' IFMBE Proceedings, Vol. 22, pp. 2007-2010

Cited by

  1. Mechano-regulation theory-based finite element analysis on the effects of driving strain history on cellular differentiation vol.16, pp.8, 2015,