Advertisement

Biomedical Microdevices

, 20:85 | Cite as

Direct application of mechanical stimulation to cell adhesion sites using a novel magnetic-driven micropillar substrate

  • Kazuaki NagayamaEmail author
  • Takuya Inoue
  • Yasuhiro Hamada
  • Shukei Sugita
  • Takeo MatsumotoEmail author
Article
  • 334 Downloads

Abstract

Cells change the traction forces generated at their adhesion sites, and these forces play essential roles in regulating various cellular functions. Here, we developed a novel magnetic-driven micropillar array PDMS substrate that can be used for the mechanical stimulation to cellular adhesion sites and for the measurement of associated cellular traction forces. The diameter, length, and center-to-center spacing of the micropillars were 3, 9, and 9 μm, respectively. Sufficient quantities of iron particles were successfully embedded into the micropillars, enabling the pillars to bend in response to an external magnetic field. We established two methods to apply magnetic fields to the micropillars (Suresh 2007). Applying a uniform magnetic field of 0.3 T bent all of the pillars by ~4 μm (Satcher et al. 1997). Creating a magnetic field gradient in the vicinity of the substrate generated a well-defined local force on the pillars. Deflection of the micropillars allowed transfer of external forces to the actin cytoskeleton through adhesion sites formed on the pillar top. Using the magnetic field gradient method, we measured the traction force changes in cultured vascular smooth muscle cells (SMCs) after local cyclic stretch stimulation at one edge of the cells. We found that the responses of SMCs were quite different from cell to cell, and elongated cells with larger pre-tension exhibited significant retraction following stretch stimulation. Our magnetic-driven micropillar substrate should be useful in investigating cellular mechanotransduction mechanisms.

Keywords

Cell biomechanics Magnetic particles Microfabrication Mechanotransduction 

Notes

Acknowledgements

This work was supported in part by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology, Japan (nos. 16 K12865 and 17H02077 to K.N., and nos.15H02209 and 17 K20102 to T.M.), Naito foundation, Japan (K.N.), Takahashi industrial and economic research foundation, Japan (K.N.), and AMED-CREST from Japan Agency for Medical Research and Development, AMED (JP18gm0810005 to K.N. and T.M.).

Supplementary material

10544_2018_328_MOESM1_ESM.pdf (89 kb)
ESM 1 (PDF 89 kb)
10544_2018_328_MOESM2_ESM.avi (47 kb)
ESM 2 (AVI 47 kb)
10544_2018_328_MOESM3_ESM.avi (55 kb)
ESM 3 (AVI 55 kb)
10544_2018_328_MOESM4_ESM.avi (25 kb)
ESM 4 (AVI 25 kb)

References

  1. Y. Arakawa, H. Bito, T. Furuyashiki, T. Tsuji, S. Takemoto-Kimura, K. Kimura, K. Nozaki, N. Hashimoto, S. Narumiya, Control of axon elongation via an SDF-1alpha/rho/mDia pathway in cultured cerebellar granule neurons. J. Cell Biol. 161(2), 381–391 (2001)CrossRefGoogle Scholar
  2. K. Burton, J.H. Park, D.L. Taylor, Keratocytes generate traction forces in two phases. Mol. Biol. Cell 10(11), 3745–3769 (1999)CrossRefGoogle Scholar
  3. C.S. Chen, M. Mrksich, S. Huang, G.M. Whitesides, D.E. Ingber, Geometric control of cell life and death. Science 276-5317, 1425–1428 (1997)CrossRefGoogle Scholar
  4. C.G. Galbraith, M.P. Sheetz, A micromachined device provides a new bend on fibroblast traction forces. Proc. Natl. Acad. Sci. U. S. A. 94(17), 9114–9118 (1997)CrossRefGoogle Scholar
  5. B. Li, M. Lin, Y. Tang, B. Wang, J.H. Wang, A novel functional assessment of the differentiation of micropatterned muscle cells. J. Biomech. 41(16), 3349–3353 (2008)CrossRefGoogle Scholar
  6. T. Mizutani, H. Haga, K. Kawabata, Cellular stiffness response to external deformation: Tensional homeostasis in a single fibroblast. Cell Motil. Cytoskeleton 59(4), 242–248 (2004)CrossRefGoogle Scholar
  7. K. Nagayama, T. Matsumoto, Dynamic change in morphology and traction forces at focal adhesions in cultured vascular smooth muscle cells during contraction. Cell. Mol. Bioeng. 4(3), 348–357 (2011)CrossRefGoogle Scholar
  8. K. Nagayama, A. Adachi, T. Matsumoto, Heterogeneous response of traction force at focal adhesions of vascular smooth muscle cells subjected to macroscopic stretch on a micropillar substrate. J. Biomech. 44(15), 2699–2705 (2011)CrossRefGoogle Scholar
  9. K. Nagayama, A. Adachi, T. Matsumoto, Dynamic changes of traction force at focal adhesions during macroscopic cell stretching using an elastic micropillar substrate: Tensional homeostasis of aortic smooth muscle cells. J. Biomed. Sci. Eng. 7(2), 130–140 (2012)Google Scholar
  10. I. Rabinovitz, I.K. Gipson, A.M. Mercurio, Traction forces mediated by alpha6beta4 integrin: Implications for basement membrane organization and tumor invasion. Mol. Biol. Cell 12(12), 4030–4043 (2001)CrossRefGoogle Scholar
  11. D. Riveline, E. Zamir, N.Q. Balaban, U.S. Schwarz, T. Ishizaki, S. Narumiya, Z. Kam, B. Geiger, A.D. Bershadsky, Focal contacts as mechanosensors: Externally applied local mechanical force induces growth of focal contacts by an mDia1-dependent and ROCK-independent mechanism. J. Cell Biol. 153(6), 1175–1186 (2001)CrossRefGoogle Scholar
  12. R. Satcher, C.F. Dewey Jr., J.H. Hartwig, Mechanical remodeling of the endothelial surface and actin cytoskeleton induced by fluid flow. Microcirculation 4, 439–453 (1997)CrossRefGoogle Scholar
  13. N.J. Sniadecki, A. Anguelouch, M.T. Yang, C.M. Lamb, Z. Liu, S.B. Kirschner, Y. Liu, D.H. Reich, C.S. Chen, Magnetic microposts as an approach to apply forces to living cells. Proc. Natl. Acad. Sci. U. S. A. 104(37), 14553–14558 (2007)CrossRefGoogle Scholar
  14. N.J. Sniadecki, C.M. Lamb, Y. Liu, C.S. Chen, D.H. Reich, Magnetic microposts for mechanical stimulation of biological cells: Fabrication, characterization, and analysis. Rev. Sci. Instrum. 79(4), 044302 (2008)CrossRefGoogle Scholar
  15. S. Sugita, T. Adachi, Y. Ueki, M. Sato, A novel method for measuring tension generated in stress fibers by applying external forces. Biophys. J. 101(1), 53–60 (2011)CrossRefGoogle Scholar
  16. S. Suresh, Biomechanics and biophysics of cancer cells. Acta Biomater. 3(4), 413–438 (2007)MathSciNetCrossRefGoogle Scholar
  17. J.L. Tan, J. Tien, D.M. Pirone, D.S. Gray, K. Bhadriraju, C.S. Chen, Cells lying on a bed of microneedles: An approach to isolate mechanical force. Proc. Natl. Acad. Sci. U. S. A. 100, 1484–1489 (2003)CrossRefGoogle Scholar
  18. Y. Ueki, N. Sakamoto, M. Sato, Cyclic force applied to FAs induces actin recruitment depending on the dynamic loading pattern. Open Biomed. Eng. J. 4, 129–134 (2010)CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Biomechanics Laboratory, Department of Mechanical EngineeringNagoya Institute of TechnologyNagoyaJapan
  2. 2.Micro-Nano Biomechanics Laboratory, Department of Mechanical Systems EngineeringIbaraki UniversityHitachiJapan
  3. 3.Biomechanics Laboratory, Department of Mechanical Systems EngineeringNagoya UniversityNagoyaJapan

Personalised recommendations