- Open Access
Energy Injection in an Epithelial Cell Monolayer Indicated by Negative Viscosity
PRX Life 3, 043017 – Published 4 December, 2025
DOI: https://doi.org/10.1103/9lnm-gm3j
Abstract
Epithelial tissues are driven out of thermodynamic equilibrium by internally generated forces, causing complex patterns of motion. Even when both the forces and motion are measurable, it is not yet possible to relate the two, because the sources of energy injection and dissipation are often unclear. Here, we study how energy is transferred by developing a method to measure the effective viscosity from the shear stresses and strain rates within an epithelial cell monolayer. Interestingly, there emerged multicellular regions in which the relationship between shear stress and shear strain rate was negatively proportional, indicating a negative effective viscosity. The negative effective viscosity occurred in regions wherein cell stresses were less efficient at producing tissue deformations compared to regions of positive effective viscosity. Regions of negative effective viscosity consistently exhibited greater cell speed and vorticity, and the cells had elevated metabolic activity, reflecting an increased energy demand in these cells. Our study shows that negative effective viscosity is a useful means of quantifying the flow of energy in living matter.
Physics Subject Headings (PhySH)
Article Text
References (58)
- M. Poujade, E. Grasland-Mongrain, A. Hertzog, J. Jouanneau, P. Chavrier, B. Ladoux, A. Buguin, and P. Silberzan, Collective migration of an epithelial monolayer in response to a model wound, Proc. Natl. Acad. Sci. USA 104, 15988 (2007).
- S. R. K. Vedula, M. C. Leong, T. L. Lai, P. Hersen, A. J. Kabla, C. T. Lim, and B. Ladoux, Emerging modes of collective cell migration induced by geometrical constraints, Proc. Natl. Acad. Sci. USA 109, 12974 (2012).
- R. Alert, J. Casademunt, and J.-F. Joanny, Active turbulence, Annu. Rev. Condens. Matter Phys. 13, 143 (2022).
- X. Trepat, M. R. Wasserman, T. E. Angelini, E. Millet, D. A. Weitz, J. P. Butler, and J. J. Fredberg, Physical forces during collective cell migration, Nat. Phys. 5, 426 (2009).
- D. T. Tambe, C. C. Hardin, T. E. Angelini, K. Rajendran, C. Y. Park, X. Serra-Picamal, E. H. Zhou, M. H. Zaman, J. P. Butler, D. A. Weitz, J. J. Fredberg, and X. Trepat, Collective cell guidance by cooperative intercellular forces, Nat. Mater. 10, 469 (2011).
- J. H. Kim, X. Serra-Picamal, D. T. Tambe, E. H. Zhou, C. Y. Park, M. Sadati, J.-A. Park, R. Krishnan, B. Gweon, E. Millet, J. P. Butler, X. Trepat, and J. J. Fredberg, Propulsion and navigation within the advancing monolayer sheet, Nat. Mater. 12, 856 (2013).
- P. K. Bera, M. McCord, J. Zhang, and J. Notbohm, Traction and stress control formation and motion of +1/2 topological defects in epithelial cell monolayers, Newton 1, 100231 (2025).
- M. C. Marchetti, J.-F. Joanny, S. Ramaswamy, T. B. Liverpool, J. Prost, M. Rao, and R. A. Simha, Hydrodynamics of soft active matter, Rev. Mod. Phys. 85, 1143 (2013).
- R. Alert and X. Trepat, Physical models of collective cell migration, Annu. Rev. Condens. Matter Phys. 11, 77 (2020).
- G. Duclos, C. Erlenkämper, J.-F. Joanny, and P. Silberzan, Topological defects in confined populations of spindle-shaped cells, Nat. Phys. 13, 58 (2017).
- C. Pérez-González, R. Alert, C. Blanch-Mercader, M. Gómez-González, T. Kolodziej, E. Bazellieres, J. Casademunt, and X. Trepat, Active wetting of epithelial tissues, Nat. Phys. 15, 79 (2019).
- P. Marmottant, A. Mgharbel, J. Käfer, B. Audren, J.-P. Rieu, J.-C. Vial, B. van der Sanden, A. F. M. Marée, F. Graner, and H. Delanoë-Ayari, The role of fluctuations and stress on the effective viscosity of cell aggregates, Proc. Natl. Acad. Sci. USA 106, 17271 (2009).
- K. Guevorkian, M.-J. Colbert, M. Durth, S. Dufour, and F. Brochard-Wyart, Aspiration of biological viscoelastic drops, Phys. Rev. Lett. 104, 218101 (2010).
- C. Blanch-Mercader, R. Vincent, E. Bazellières, X. Serra-Picamal, X. Trepat, and J. Casademunt, Effective viscosity and dynamics of spreading epithelia: A solvable model, Soft Matter 13, 1235 (2017).
- J. Notbohm, S. Banerjee, K. J. Utuje, B. Gweon, H. Jang, Y. Park, J. Shin, J. P. Butler, J. J. Fredberg, and M. C. Marchetti, Cellular contraction and polarization drive collective cellular motion, Biophys. J. 110, 2729 (2016).
- A. Saraswathibhatla and J. Notbohm, Tractions and stress fibers control cell shape and rearrangements in collective cell migration, Phys. Rev. X 10, 011016 (2020).
- D. T. Tambe, U. Croutelle, X. Trepat, C. Y. Park, J. H. Kim, E. Millet, J. P. Butler, and J. J. Fredberg, Monolayer stress microscopy: Limitations, artifacts, and accuracy of recovered intercellular stresses, PLoS ONE 8, e55172 (2013).
- A. Saraswathibhatla, E. E. Galles, and J. Notbohm, Spatiotemporal force and motion in collective cell migration, Sci. Data 7, 197 (2020).
- Y. Hatwalne, S. Ramaswamy, M. Rao, and R. A. Simha, Rheology of active-particle suspensions, Phys. Rev. Lett. 92, 118101 (2004).
- M. E. Cates, S. M. Fielding, D. Marenduzzo, E. Orlandini, and J. M. Yeomans, Shearing active gels close to the isotropic-nematic transition, Phys. Rev. Lett. 101, 068102 (2008).
- A. Sokolov and I. S. Aranson, Reduction of viscosity in suspension of swimming bacteria, Phys. Rev. Lett. 103, 148101 (2009).
- J. Gachelin, G. Mino, H. Berthet, A. Lindner, A. Rousselet, and É. Clément, Non-newtonian viscosity of Escherichia coli suspensions, Phys. Rev. Lett. 110, 268103 (2013).
- H. M. López, J. Gachelin, C. Douarche, H. Auradou, and E. Clément, Turning bacteria suspensions into superfluids, Phys. Rev. Lett. 115, 028301 (2015).
- H. Orihara, Y. Harada, F. Kobayashi, Y. Sasaki, S. Fujii, Y. Satou, Y. Goto, and T. Nagaya, Negative viscosity of a liquid crystal in the presence of turbulence, Phys. Rev. E 99, 012701 (2019).
- J. Y. Chui, C. Douarche, H. Auradou, and R. Juanes, Rheology of bacterial superfluids in viscous environments, Soft Matter 17, 7004 (2021).
- M. McCord and J. Notbohm, Measurement of tissue viscosity to relate force and motion in collective cell migration, arXiv:2508.11518.
- D. Saintillan, Rheology of active fluids, Annu. Rev. Fluid Mech. 50, 563 (2018).
- M. R. Nejad, L. J. Ruske, M. McCord, J. Zhang, G. Zhang, J. Notbohm, and J. M. Yeomans, Stress-shape misalignment in confluent cell layers, Nat. Commun. 15, 3628 (2024).
- P. Lee and C. Wolgemuth, Advent of complex flows in epithelial tissues, Phys. Rev. E 83, 061920 (2011).
- O. Cochet-Escartin, J. Ranft, P. Silberzan, and P. Marcq, Border forces and friction control epithelial closure dynamics, Biophys. J. 106, 65 (2014).
- K. Vazquez, A. Saraswathibhatla, and J. Notbohm, Effect of substrate stiffness on friction in collective cell migration, Sci. Rep. 12, 2436 (2022).
- S.-Z. Lin, W.-Y. Zhang, D. Bi, B. Li, and X.-Q. Feng, Energetics of mesoscale cell turbulence in two-dimensional monolayers, Commun. Phys. 4, 21 (2021).
- R. C. Scaduto and L. W. Grotyohann, Measurement of mitochondrial membrane potential using fluorescent rhodamine derivatives, Biophys. J. 76, 469 (1999).
- C. Zou, Y. Wang, and Z. Shen, 2-NBDG as a fluorescent indicator for direct glucose uptake measurement, J. Biochem. Biophys. Methods 64, 207 (2005).
- S. J. DeCamp, V. M. K. Tsuda, J. Ferruzzi, S. A. Koehler, J. T. Giblin, D. Roblyer, M. H. Zaman, S. T. Weiss, A. Kılıç, M. De Marzio, C. Y. Park, N. C. Ogassavara, J. A. Mitchel, J. P. Butler, and J. J. Fredberg, Epithelial layer unjamming shifts energy metabolism toward glycolysis, Sci. Rep. 10, 18302 (2020).
- S. Baron, A. Caplanusi, M. van de Ven, M. Radu, S. Despa, I. Lambrichts, M. Ameloot, P. Steels, and I. Smets, Role of mitochondrial concentration, measured by CoroNa red, in the protection of metabolically inhibited MDCK cells, J. Am. Soc. Nephrol. 16, 3490 (2005).
- A. J. Walsh, K. Mueller, K. Tweed, I. Jones, C. M. Walsh, N. Piscopo, N. M. Niemi, D. J. Pagliarini, K. Saha, and M. C. Skala, Classification of t-cell activation via autofluorescence lifetime imaging, Nat. Biomed. Eng. 5, 77 (2021).
- M. J. Filon, A. A. Gillette, B. Yang, T. A. Khemees, M. C. Skala, and D. F. Jarrard, Prostate cancer cells demonstrate unique metabolism and substrate adaptability acutely after androgen deprivation therapy, Prostate 82, 1547 (2022).
- A. Saraswathibhatla, S. Henkes, E. E. Galles, R. Sknepnek, and J. Notbohm, Coordinated tractions increase the size of a collectively moving pack in a cell monolayer, Extreme Mech. Lett. 48, 101438 (2021).
- R. Lakes and W. Drugan, Dramatically stiffer elastic composite materials due to a negative stiffness phase? J. Mech. Phys. Solids 50, 979 (2002).
- T. Jaglinski, D. Kochmann, D. Stone, and R. S. Lakes, Composite materials with viscoelastic stiffness greater than diamond, Science 315, 620 (2007).
- H. H. Wensink, J. Dunkel, S. Heidenreich, K. Drescher, R. E. Goldstein, H. Löwen, and J. M. Yeomans, Meso-scale turbulence in living fluids, Proc. Natl. Acad. Sci. USA 109, 14308 (2012).
- N. S. Rossen, J. M. Tarp, J. Mathiesen, M. H. Jensen, and L. B. Oddershede, Long-range ordered vorticity patterns in living tissue induced by cell division, Nat. Commun. 5, 5720 (2014).
- A. G. Smart, L. A. Amaral, and J. M. Ottino, Cascading failure and robustness in metabolic networks, Proc. Natl. Acad. Sci. USA 105, 13223 (2008).
- J. Stelling, U. Sauer, Z. Szallasi, F. J. Doyle, and J. Doyle, Robustness of cellular functions, Cell 118, 675 (2004).
- J. M. Whitacre, Biological robustness: Paradigms, mechanisms, and systems principles, Front Genet. 3, 67 (2012).
- R. J. DeBerardinis and N. S. Chandel, Fundamentals of cancer metabolism, Sci. Adv. 2, e1600200 (2016).
- G. Bergers and S.-M. Fendt, The metabolism of cancer cells during metastasis, Nat. Rev. Cancer 21, 162 (2021).
- Z. Wang, F. Zhao, C. Xu, Q. Zhang, H. Ren, X. Huang, C. He, J. Ma, and Z. Wang, Metabolic reprogramming in skin wound healing, Burns Trauma 12, tkad047 (2024).
- R. Iwata and P. Vanderhaeghen, Metabolic mechanisms of species-specific developmental tempo, Dev. Cell 59, 1628 (2024).
- E. Bar-Kochba, J. Toyjanova, E. Andrews, K.-S. Kim, and C. Franck, A fast iterative digital volume correlation algorithm for large deformations, Exp. Mech. 55, 261 (2015).
- J. P. Butler, I. M. Tolic-Norrelykke, B. Fabry, and J. J. Fredberg, Traction fields, moments, and strain energy that cells exert on their surroundings, Am. J. Physiol. Cell Physiol. 282, C595 (2002).
- J. C. del Alamo, R. Meili, B. Alonso-Latorre, J. Rodriguez-Rodriguez, A. Aliseda, R. A. Firtel, and J. C. Lasheras, Spatio-temporal analysis of eukaryotic cell motility by improved force cytometry, Proc. Natl. Acad. Sci. USA 104, 13343 (2007).
- J. A. Vromans and L. Giomi, Orientational properties of nematic disclinations, Soft Matter 12, 6490 (2016).
- I. Smets, A. Caplanusi, S. Despa, Z. Molnar, M. Radu, M. vandeVen, M. Ameloot, and P. Steels, uptake in mitochondria occurs via the reverse action of the exchanger in metabolically inhibited MDCK cells, Am. J. Physiol. Renal. Physiol. 286, F784 (2004).
- https://github.com/jknotbohm/FIDIC.
- https://github.com/jknotbohm/Cell-Traction-Stress.
- D. Bi, X. Yang, M. C. Marchetti, and M. L. Manning, Motility-driven glass and jamming transitions in biological tissues, Phys. Rev. X 6, 021011 (2016).