Mechanical properties of cultured human airway smooth muscle cells from 0.05 to 0.4 Hz.

We investigated the rheological properties of living human airway smooth muscle cells in culture and monitored the changes in rheological properties induced by exogenous stimuli. We oscillated small magnetic microbeads bound specifically to integrin receptors and computed the storage modulus (G') and loss modulus (G") from the applied torque and the resulting rotational motion of the beads as determined from their remanent magnetic field. Under baseline conditions, G' increased weakly with frequency, whereas G" was independent of the frequency. The cell was predominantly elastic, with the ratio of G" to G' (defined as eta) being approximately 0. 35 at all frequencies. G' and G" increased together after contractile activation and decreased together after deactivation, whereas eta remained unaltered in each case. Thus elastic and dissipative stresses were coupled during changes in contractile activation. G' and G" decreased with disruption of the actin fibers by cytochalasin D, but eta increased. These results imply that the mechanisms for frictional energy loss and elastic energy storage in the living cell are coupled and reside within the cytoskeleton.

[1]  R. M. Simmons,et al.  Elasticity and unfolding of single molecules of the giant muscle protein titin , 1997, Nature.

[2]  Donald E. Ingber,et al.  Integrin binding and mechanical tension induce movement of mRNA and ribosomes to focal adhesions , 1998, Nature.

[3]  Mario Viani,et al.  Molecular mechanistic origin of the toughness of natural adhesives, fibres and composites , 1999, Nature.

[4]  L. E. Ford,et al.  Plasticity in canine airway smooth muscle , 1995, The Journal of general physiology.

[5]  P. Gennes Reptation of a Polymer Chain in the Presence of Fixed Obstacles , 1971 .

[6]  S Chien,et al.  Shear stress induces spatial reorganization of the endothelial cell cytoskeleton. , 1998, Cell motility and the cytoskeleton.

[7]  Francis Crick,et al.  The physical properties of cytoplasm. A study by means of the magnetic particle method. Part II. Theoretical treatment , 1950 .

[8]  R. Buxbaum,et al.  F-actin and microtubule suspensions as indeterminate fluids. , 1987, Science.

[9]  M Grattarola,et al.  Mechanical and morphological properties of living 3T6 cells probed via scanning force microscopy , 1997, Microscopy research and technique.

[10]  U G Hofmann,et al.  Investigating the cytoskeleton of chicken cardiocytes with the atomic force microscope. , 1997, Journal of structural biology.

[11]  M. Huggins Viscoelastic Properties of Polymers. , 1961 .

[12]  J. Fredberg,et al.  Airway smooth muscle, tidal stretches, and dynamically determined contractile states. , 1997, American journal of respiratory and critical care medicine.

[13]  J. Fredberg,et al.  Perturbed equilibrium of myosin binding in airway smooth muscle and its implications in bronchospasm. , 1999, American journal of respiratory and critical care medicine.

[14]  S. Gunst,et al.  Mechanisms for the mechanical plasticity of tracheal smooth muscle. , 1995, The American journal of physiology.

[15]  S. Edwards,et al.  The Theory of Polymer Dynamics , 1986 .

[16]  J J Fredberg,et al.  Pharmacological activation changes stiffness of cultured human airway smooth muscle cells. , 1996, The American journal of physiology.

[17]  D Lerche,et al.  The mechanical properties of actin gels. Elastic modulus and filament motions. , 1994, The Journal of biological chemistry.

[18]  S. H. Crandall The role of damping in vibration theory , 1970 .

[19]  M Nathan,et al.  Friction in airway smooth muscle: mechanism, latch, and implications in asthma. , 1996, Journal of applied physiology.

[20]  P. Valberg Magnetometry of ingested particles in pulmonary macrophages. , 1984, Science.

[21]  Y. Wang,et al.  Preparation of a flexible, porous polyacrylamide substrate for mechanical studies of cultured cells. , 1998, Methods in enzymology.

[22]  R Ezzell,et al.  F-actin, a model polymer for semiflexible chains in dilute, semidilute, and liquid crystalline solutions. , 1996, Biophysical journal.

[23]  D Stamenović,et al.  The role of prestress and architecture of the cytoskeleton and deformability of cytoskeletal filaments in mechanics of adherent cells: a quantitative analysis. , 1999, Journal of theoretical biology.

[24]  E. Sackmann,et al.  Local measurements of viscoelastic moduli of entangled actin networks using an oscillating magnetic bead micro-rheometer. , 1994, Biophysical journal.

[25]  D. Stamenović,et al.  On the imperfect elasticity of lung tissue. , 1989, Journal of applied physiology.

[26]  Francis Crick,et al.  The physical properties of cytoplasm: A study by means of the magnetic particle method Part I. Experimental , 1950 .

[27]  K. Jacobson,et al.  Local measurements of viscoelastic parameters of adherent cell surfaces by magnetic bead microrheometry. , 1998, Biophysical journal.

[28]  P. Janmey,et al.  Mechanical perturbation elicits a phenotypic difference between Dictyostelium wild-type cells and cytoskeletal mutants. , 1996, Biophysical journal.

[29]  Robert E. Buxbaum,et al.  Direct Observations of the Mechanical Behaviors of the Cytoskeleton in Living Fibroblasts , 1999, The Journal of cell biology.

[30]  P A Valberg,et al.  Magnetic particle motions within living cells. Physical theory and techniques. , 1987, Biophysical journal.

[31]  N O Petersen,et al.  Dependence of locally measured cellular deformability on position on the cell, temperature, and cytochalasin B. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[32]  W. H. Goldmann,et al.  Viscoelasticity in wild-type and vinculin-deficient (5.51) mouse F9 embryonic carcinoma cells examined by atomic force microscopy and rheology. , 1996, Experimental cell research.

[33]  Jean-Jacques Meister,et al.  Changes in the mechanical properties of fibroblasts during spreading: a micromanipulation study , 1999, European Biophysics Journal.

[34]  R M Nerem,et al.  Viscoelastic properties of cultured porcine aortic endothelial cells exposed to shear stress. , 1996, Journal of biomechanics.

[35]  J. Butler,et al.  Intracellular elasticity and viscosity in the body , leading , and trailing regions of locomoting neutrophils , 1999 .

[36]  T. Stossel On the crawling of animal cells. , 1993, Science.

[37]  J. Butler,et al.  Cytoskeletal mechanics in confluent epithelial cells probed through integrins and E-cadherins. , 1997, The American journal of physiology.

[38]  B. Nebe,et al.  Stimulation of integrin receptors using a magnetic drag force device induces an intracellular free calcium response. , 1996, European journal of cell biology.

[39]  G. Shue,et al.  The frequency response of smooth muscle stiffness during Ca2+-activated contraction. , 1999, Biophysical journal.

[40]  P. Janmey,et al.  Mechanical Effects of Neurofilament Cross-bridges , 1996, The Journal of Biological Chemistry.

[41]  D. Ingber,et al.  Probing transmembrane mechanical coupling and cytomechanics using magnetic twisting cytometry. , 1995, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[42]  R. Lal,et al.  Dynamic micromechanical properties of cultured rat atrial myocytes measured by atomic force microscopy. , 1995, The American journal of physiology.

[43]  C. Hirshman,et al.  Actin reorganization in airway smooth muscle cells involves Gq and Gi-2 activation of Rho. , 1999, The American journal of physiology.

[44]  G. Forgacs On the possible role of cytoskeletal filamentous networks in intracellular signaling: an approach based on percolation. , 1995, Journal of cell science.

[45]  M. Sato [Mechanical properties of living tissues]. , 1986, Iyo denshi to seitai kogaku. Japanese journal of medical electronics and biological engineering.

[46]  C. S. Chen,et al.  Geometric control of cell life and death. , 1997, Science.

[47]  R. Panettieri,et al.  A human airway smooth muscle cell line that retains physiological responsiveness. , 1989, The American journal of physiology.

[48]  D. Rees,et al.  Characterization of cross-bridge elasticity and kinetics of cross-bridge cycling during force development in single smooth muscle cells , 1988, The Journal of general physiology.

[49]  D. Ingber Tensegrity: the architectural basis of cellular mechanotransduction. , 1997, Annual review of physiology.

[50]  R. Skalak,et al.  Passive mechanical properties of human leukocytes. , 1981, Biophysical Journal.

[51]  D. Ingber,et al.  Integrins as mechanochemical transducers. , 1991, Current opinion in cell biology.

[52]  D. Ingber,et al.  Mechanotransduction across the cell surface and through the cytoskeleton , 1993 .

[53]  E. Sackmann,et al.  Measurement of local viscoelasticity and forces in living cells by magnetic tweezers. , 1999, Biophysical journal.

[54]  Ben Fabry,et al.  Implications of heterogeneous bead behavior on cell mechanical properties measured with magnetic twisting cytometry , 1999 .

[55]  D E Ingber,et al.  Convergence of integrin and growth factor receptor signaling pathways within the focal adhesion complex. , 1995, Molecular biology of the cell.

[56]  J. Bereiter-Hahn,et al.  Mechanical basis of cell shape: investigations with the scanning acoustic microscope. , 1995, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[57]  W. Goldmann,et al.  Viscoelasticity of actin-gelsolin networks in the presence of filamin. , 1997, European journal of biochemistry.

[58]  D E Ingber,et al.  Control of cytoskeletal mechanics by extracellular matrix, cell shape, and mechanical tension. , 1994, Biophysical journal.

[59]  A. Palmer,et al.  Diffusing wave spectroscopy microrheology of actin filament networks. , 1999, Biophysical journal.