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1 Physiology Program, Harvard School of Public Health, Boston, Massachusetts 02115; 2 School of Biomedical Engineering, Dalhousie University, Halifax, Nova Scotia, Canada B3H 3J5; and 3 Pulmonary and Critical Care Division, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104
We measured the time course and heterogeneity of responses to
contractile and relaxing agonists in individual human airway smooth
muscle (HASM) cells in culture. To this end, we developed a
microrheometer based on magnetic twisting cytometry adapted with a
novel optical detection system. Ferromagnetic beads (4.5 µm) coated
with Arg-Gly-Asp peptide were bound to integrins on the cell surface.
The beads were twisted in a sinusoidally varying magnetic field at 0.75 Hz. Oscillatory bead displacements were recorded using a
phase-synchronized video camera. The storage modulus (cell stiffness;
G'), loss modulus (friction; G"), and hysteresivity (
; ratio of G"
to G') could be determined with a time resolution of 1.3 s. Within
5 s after addition of histamine (100 µM), G' increased by
2.2-fold, G" increased by 3.0-fold, and increased transiently from
0.27 to 0.34. By 20 s, decreased to 0.25, whereas G' and G"
remained above baseline. Comparable results were obtained with
bradykinin (1 µM). These changes in G', G", and measured in cells
were similar to but smaller than those reported for intact muscle
strips. When we ablated baseline tone by adding the relaxing agonist
dibutyryl cAMP (1 mM), G' decreased within 5 min by 3.3-fold. With
relaxing and contracting agonists, G' could be manipulated through a
contractile range of 7.3-fold. Cell populations exhibited a log-normal
distribution of baseline stiffness (geometric SD = 2.8) and a
heterogeneous response to both contractile and relaxing agonists,
partly attributable to variability of baseline tone between cells. The
total contractile range of the cells (from maximally relaxed to
maximally stimulated), however, was independent of baseline stiffness.
We conclude that HASM cells in culture exhibit a clear, although
heterogeneous, response to contractile and relaxing agonists and
express the essential mechanical features characteristic of the
contractile response observed at the tissue level.
cell mechanics; muscle contraction; muscle relaxation; actin; myosin; bridge dynamics; cytoskeleton
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