Abstract
The cytoskeleton is primarily responsible for providing structural support, localization and transport of organelles, and intracellular trafficking. The structural support is supplied by actin filaments, microtubules, and intermediate filaments, which contribute to overall cell elasticity to varying degrees. We evaluate cell elasticity in five different cell types with drug-induced cytoskeletal derangements to probe how actin filaments and microtubules contribute to cell elasticity and whether it is conserved across cell type. Specifically, we measure elastic stiffness in chondrocytes, fibroblasts, endothelial cells, hepatocellular carcinoma, and fibrosarcoma using atomic force microscopy. We subject all five cell lines to two cytoskeletal destabilizers: cytochalasin D and nocodazole, which disrupt actin and microtubule polymerization, respectively. Non-cancer cells treated with cytochalasin D show a decrease of 60–80 % in moduli values compared to untreated cells of the same origin, whereas the nocodazole-treated cells show no change. Alternatively, cancer cells exhibit increased stiffness as well as stiffness variability when subjected to nocodazole. Overall, we demonstrate actin filaments contribute more to elastic stiffness than microtubules but this result is cell type dependent. Lastly, we show that disruption of microtubule dynamics affects cancer cell elasticity, suggesting therapeutic drugs targeting microtubules be monitored for significant elastic changes.
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References
Discher, D.E., Janmey, P., Wang, Y.L.: Tissue cells feel and respond to the stiffness of their substrate. Science 310(5751), 1139–1143 (2005)
Bao, G., Suresh, S.: Cell and molecular mechanics of biological materials. Nat. Mater. 2(11), 715–725 (2003)
Lee, G.Y.H., Lim, C.T.: Biomechanics approaches to studying human diseases. Trends Biotechnol. 25(3), 111–118 (2007)
Rother, J., Noding, H., Mey, I., Janshoff, A.: Atomic force microscopy-based microrheology reveals significant differences in the viscoelastic response between malign and benign cell lines. Open Biol. 4(5), 140046 (2014)
Nalam, P.C., Gosvami, N.N., Caporizzo, M.A., Composto, R.J., Carpick, R.W.: Nano-rheology of hydrogels using direct drive force modulation atomic force microscopy. Soft Matter 11(41), 8165–8178 (2015)
Raman, A., Trigueros, S., Cartagena, A., Stevenson, A.P.Z., Susilo, M., Nauman, E., Contera, S.A.: Mapping nanomechanical properties of live cells using multi-harmonic atomic force microscopy. Nat. Nanotechnol. 6(12), 809–814 (2011)
Caporizzo, M.A., Roco, C.M., Ferrer, M.C.C., Grady, M.E., Parrish, E., Eckmann, D.M., Composto, R.J.: Strain-rate dependence of elastic modulus reveals silver nanoparticle induced cytotoxicity. Nanobiomedicine 2 (2015). pii: 9
Ketene, A.N., Schmelz, E.M., Roberts, P.C., Agah, M.: The effects of cancer progression on the viscoelasticity of ovarian cell cytoskeleton structures. Nanomedicine 8(1), 93–102 (2012)
Corbin, E.A., Kong, F., Lim, C.T., King, W.P., Bashir, R.: Biophysical properties of human breast cancer cells measured using silicon MEMS resonators and atomic force microscopy. Lab Chip 15(3), 839–847 (2014)
Moreno-Flores, S., Benitez, R., Vivanco, M.D., Toca-Herrera, J.L.: Stress relaxation and creep on living cells with the atomic force microscope: a means to calculate elastic moduli and viscosities of cell components. Nanotechnology 21(44), 445101 (2010)
Kuznetsova, T.G., Starodubtseva, M.N., Yegorenkov, N.I., Chizhik, S.A., Zhdanov, R.I.: Atomic force microscopy probing of cell elasticity. Micron 38(8), 824–833 (2007)
Rebelo, L.M., de Sousa, J.S., Mendes, J., Radmacher, M.: Comparison of the viscoelastic properties of cells from different kidney cancer phenotypes measured with atomic force microscopy. Nanotechnology 24(5), 055102 (2013)
Park, S., Koch, D., Cardenas, R., Kas, J., Shih, C.K.: Cell motility and local viscoelasticity of fibroblasts. Biophys. J. 89(6), 4330–4342 (2005)
Ribeiro, A.S., Khanna, P., Sukumar, A., Dong, C., Dahl, K.: Nuclear stiffening inhibits migration of invasive melanoma cells. Cell. Mol. Bioeng. 7(4), 544–551 (2014)
Buda, A., Pignatelli, M.: Cytoskeletal network in colon cancer: from genes to clinical application. Int. J. Biochem. Cell Biol. 36(5), 759–765 (2004)
Lindberg, U., Karlsson, R., Lassing, I., Schutt, C.E., Höglund, A.-S.: The microfilament system and malignancy. Semin. Cancer Biol. 18(1), 2–11 (2008)
Grady, M.E., Composto, R.J., Eckmann, D.M.: Cell elasticity with altered cytoskeletal architectures across multiple cell types. J. Mech. Behav. Biomed. Mater. 61, 197–207 (2016)
Trickey, W.R., Baaijens, F.P.T., Laursen, T.A., Alexopoulos, L.G., Guilak, F.: Determination of the Poisson's ratio of the cell: recovery properties of chondrocytes after release from complete micropipette aspiration. J. Biomech. 39(1), 78–87 (2006)
Acknowledgements
The authors gratefully acknowledge our funding sources: ONR Grant N000141612100 (DME), the Provost’s Postdoctoral Fellowship for Academic Diversity (MEG), NSF-NSEC Grant DMR08-32802 (RJC), and URF 4-000002-4820 (DME), which made this work possible. The work was performed at and supported by the Nano Bio Interface Center at the University of Pennsylvania through an instrumentation grant, DBI-0721913, and DMR-0425780. We also thank Judith Kandel for cell culture training and Dr. Matt Brukman and Dr. Matt Caporizzo for instrument support. We thank the following for their generous donations: Dr. Robert Mauck (chondrocytes), Dr. Ben Stanger (HUH-7), and Dr. Bruce Malkowicz (HT-1080).
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Grady, M.E., Composto, R.J., Eckmann, D.M. (2017). Cytoskeletal Perturbing Drugs and Their Effect on Cell Elasticity. In: Korach, C., Tekalur, S., Zavattieri, P. (eds) Mechanics of Biological Systems and Materials, Volume 6. Conference Proceedings of the Society for Experimental Mechanics Series. Springer, Cham. https://doi.org/10.1007/978-3-319-41351-8_24
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DOI: https://doi.org/10.1007/978-3-319-41351-8_24
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