Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Macrophage adhesion on fibronectin evokes an increase in the elastic property of the cell membrane and cytoskeleton: an atomic force microscopy study

  • 683 Accesses

  • 11 Citations


Interactions between cells and microenvironments are essential to cellular functions such as survival, exocytosis and differentiation. Cell adhesion to the extracellular matrix (ECM) evokes a variety of biophysical changes in cellular organization, including modification of the cytoskeleton and plasma membrane. In fact, the cytoskeleton and plasma membrane are structures that mediate adherent contacts with the ECM; therefore, they are closely correlated. Considering that the mechanical properties of the cell could be affected by cell adhesion-induced changes in the cytoskeleton, the purpose of this study was to investigate the influence of the ECM on the elastic properties of fixed macrophage cells using atomic force microscopy. The results showed that there was an increase (~50 %) in the Young’s modulus of macrophages adhered to an ECM-coated substrate as compared with an uncoated glass substrate. In addition, cytochalasin D-treated cells had a 1.8-fold reduction of the Young’s modulus of the cells, indicating the contribution of the actin cytoskeleton to the elastic properties of the cell. Our findings show that cell adhesion influences the mechanical properties of the plasma membrane, providing new information toward understanding the influence of the ECM on elastic alterations of macrophage cell membranes.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5


  1. Binnig G, Quate CF, Gerber C (1986) Atomic force microscope. Phys Rev Lett 56:930–933

  2. Bukharaev AA, Mozhanova AA, Nurgazizov NI, Ovchinnikov DV (2003) Measuring local elastic properties of cell surfaces and soft materials in liquid by atomic force microscopy. Phys Low-Dimens Str 3–4:31–37

  3. Burridge K, Chrzanowska-Wodnicka M (1996) Focal adhesions, contractility, and signaling. Annu Rev Cell Dev Bi 12:463–518

  4. Bushell GR, Cahill C, Clarke FM, Gibson CT, Myhra S, Watson GS (1999) Imaging and force-distance analysis of human fibroblasts in vitro by atomic force microscopy. Cytometry 36:254–264

  5. Cappella B, Baschieri P, Frediani C, Miccoli P, Ascoli C (1997) Force-distance curves by AFM. A powerful technique for studying surface interactions. IEEE Eng Med Biol Mag 16:58–65

  6. Codan B, Martinelli V, Mestroni L, Sbaizero O (2013) Atomic force microscopy of 3T3 and SW-13 cell lines: an investigation of cell elasticity changes due to fixation. Mat Sci Eng C-Mater 33:3303–3308

  7. Collins SJ (1987) The Hl-60 promyelocytic leukemia-cell line—proliferation, differentiation, and cellular oncogene expression. Blood 70:1233–1244

  8. Dai JW, Sheetz MP (1999) Membrane tether formation from blebbing cells. Biophys J 77:3363–3370

  9. Fischer-Cripps AC (2011) Nanoindentation, 3rd edn. Springer, New York

  10. Gauthier NC, Masters TA, Sheetz MP (2012) Mechanical feedback between membrane tension and dynamics. Trends Cell Biol 22:527–535

  11. Gordon S (2003) Alternative activation of macrophages. Nat Rev Immunol 3:23–35

  12. Gordon S, Martinez FO (2010) Alternative activation of macrophages: mechanism and functions. Immunity 32:593–604

  13. Hemler ME (1990) Vla proteins in the integrin family—structures, functions, and their role on leukocytes. Annu Rev Immunol 8:365–400

  14. Hertz H (1881) Ueber den kontakt elastischer koerper. J Reine Angew Math 92:156–171

  15. Hochmuth RM, Shao JY, Dai JW, Sheetz MP (1996) Deformation and flow of membrane into tethers extracted from neuronal growth cones. Biophys J 70:358–369

  16. Horcas I, Fernandez R, Gomez-Rodriguez JM, Colchero J, Gomez- Herrero J, Baro AM (2007) WSXM: A software for scanning probe microscopy and a tool for nanotechnology. Rev Sci Instrum 78

  17. Ingber DE, Dike L, Hansen L, Karp S, Liley H, Maniotis A, Mcnamee H, Mooney D, Plopper G, Sims J, Wang N (1994) Cellular tensegrity—exploring how mechanical changes in the cytoskeleton regulate cell-growth, migration, and tissue pattern during morphogenesis. Int Rev Cytol 150:173–224

  18. Janmey PA (1998) The cytoskeleton and cell signaling: component localization and mechanical coupling. Physiol Rev 78:763–781

  19. Johnson KL (1985) Contact mechanics. Cambridge University Press, Cambridge

  20. Kakaboura A, Fragouli M, Rahiotis C, Silikas N (2007) Evaluation of surface characteristics of dental composites using profilometry, scanning electron, atomic force microscopy and gloss-meter. J Mater Sci-Mater M 18:155–163

  21. Keren K (2011) Cell motility: the integrating role of the plasma membrane. Eur Biophys J Biophy 40:1013–1027

  22. Kreider T, Anthony RM, Urban JF, Gause WC (2007) Alternatively activated macrophages in helminth infections. Curr Opin Immunol 19:448–453

  23. Lee YJ, Patel D, Park S (2011) Local rheology of human neutrophils investigated using atomic force microscopy. Int J Biol Sci 7:102–111

  24. Lekka M, Gil D, Pogoda K, Dulinska-Litewka J, Jach R, Gostek J, Klymenko O, Prauzner-Bechcicki S, Stachura Z, Wiltowska-Zuber J, Okon K, Laidler P (2012) Cancer cell detection in tissue sections using AFM. Arch Biochem Biophys 518:151–156

  25. Mackay JL, Kumar S (2013) Measuring the elastic properties of living cells with atomic force microscopy indentation. Methods Mol Biol 931:313–329

  26. Mahaffy RE, Park S, Gerde E, Kas J, Shih CK (2004) Quantitative analysis of the viscoelastic properties of thin regions of fibroblasts using atomic force microscopy. Biophys J 86:1777–1793

  27. Maniotis AJ, Chen CS, Ingber DE (1997) Demonstration of mechanical connections between integrins cytoskeletal filaments, and nucleoplasm that stabilize nuclear structure. Proc Natl Acad Sci USA 94:849–854

  28. Morris CE, Homann U (2001) Cell surface area regulation and membrane tension. J Membrane Biol 179:79–102

  29. Mozhanova AA, Nurgazizov NI, Bukharaev AA(2003) Local elastic properties of biological materials studied by SFM. SPM-2003 In: Proceedings, Nizhni Novgorod, pp 266–267

  30. Patel NR, Bole M, Chen C, Hardin CC, Kho AT, Mih J, Deng LH, Butler J, Tschumperlin D, Fredberg JJ, Krishnan R, Koziel H (2012) Cell elasticity determines macrophage function. PLoS One 7(9):e41024

  31. Radmacher M (1997) Measuring the elastic properties of biological samples with the AFM. Ieee Eng Med Biol 16:47–57

  32. Radmacher M, Fritz M, Hansma PK (1995) Imaging soft samples with the atomic-force microscope—gelatin in water and propanol. Biophys J 69:264–270

  33. Raman A, Trigueros S, Cartagena A, Stevenson APZ, Susilo M, Nauman E, Contera SA (2011) Mapping nanomechanical properties of live cells using multi-harmonic atomic force microscopy. Nat Nanotechnol 6:809–814

  34. Rotsch C, Braet F, Wisse E, Radmacher M (1997) AFM imaging and elasticity measurements on living rat liver macrophages. Cell Biol Int 21:685–696

  35. Segat D, Pucillo C, Marotta G, Perris R, Colombatti A (1994) Differential attachment of human neoplastic B-cells to purified extracellular-matrix molecules. Blood 83:1586–1594

  36. Sheetz MP, Dai JW (1996) Modulation of membrane dynamics and cell motility by membrane tension. Trends Cell Biol 6:85–89

  37. Shroff SG, Saner DR, Lal R (1995) Dynamic micromechanical properties of cultured rat atrial myocytes measured by atomic-force microscopy. Am J Physiol 269:C286–C292

  38. Sirghi L, Ponti J, Broggi F, Rossi F (2008) Probing elasticity and adhesion of live cells by atomic force microscopy indentation. Eur Biophys J 37:935–945

  39. Sneddon IN (1965) The relation between load and penetration in the axisymmetric boussinesq problem for a punch of arbitrary profile. Int J Eng Sci 3:47–57

  40. Spedden E, White JD, Naumova EN, Kaplan DL, Staii C (2012) Elasticity maps of living neurons measured by combined fluorescence and atomic force microscopy. Biophys J 103:868–877

  41. Svaldo-Lanero T, Krol S, Magrassi R, Diaspro A, Rolandi R, Gliozzi A, Cavalleri O (2007) Morphology, mechanical properties and viability of encapsulated cells. Ultramicroscopy 107:913–921

  42. Vesey DA, Cheung CWY, Cuttle L, Endre ZA, Gobe G, Johnson DW (2002) Interleukin-1 beta induces human proximal tubule cell injury, alpha-smooth muscle actin expression and fibronectin production. Kidney Int 62:31–40

  43. Weisenhorn AL, Hansma PK, Albrecht TR, Quate CF (1989) Forces in atomic force microscopy in air and water. Appl Phys Lett 54:2651–2653

  44. Wu HW, Kuhn T, Moy VT (1998) Mechanical properties of l929 cells measured by atomic force microscopy: effects of anticytoskeletal drugs and membrane crosslinking. Scanning 20:389–397

Download references


This research was supported by CAPES/Nanobiotecnologia, Pró-equipamentos/PROCAD, Pronex/FAPEAL, CNPq and FINEP.

Author information

Correspondence to Eduardo J. S. Fonseca.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Souza, S.T., Agra, L.C., Santos, C.E.A. et al. Macrophage adhesion on fibronectin evokes an increase in the elastic property of the cell membrane and cytoskeleton: an atomic force microscopy study. Eur Biophys J 43, 573–579 (2014). https://doi.org/10.1007/s00249-014-0988-3

Download citation


  • Atomic force microscopy
  • Elastic modulus
  • Nanoindentation
  • Extracellular matrix
  • Cytoskeleton
  • Fibronectin