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HIGHLIGHTED TOPICS
Biomechanics and Mechanotransduction in Cells and Tissues

Thrombin and histamine induce stiffening of alveolar epithelial cells

Unitat de Biofísica i Bioenginyeria, Facultat de Medicina, Universitat de Barcelona-Institut d’Investigacions Biomèdiques August Pi i Sunyer, Barcelona, Spain

Submitted 26 August 2004 ; accepted in final form 11 November 2004

The mechanical properties of alveolar epithelial cells play a central role in maintaining the physical integrity of the alveolar epithelium. We studied the viscoelastic properties of alveolar epithelial cells (A549) in response to thrombin and histamine with optical magnetic twisting cytometry. Ferrimagnetic beads coated with Arg-Gly-Asp (RGD)-peptide or acetylated low-density lipoprotein were bound to cell surface receptors and subsequently twisted in an oscillatory magnetic field (0.1–100 Hz). The cell storage (G') and loss (G'') moduli were computed from twisting torque and bead displacement. In measurements with RGD-coated beads, thrombin (0.5 U/ml) induced a rapid and sustained threefold increase in G' and G'' at 100 s after challenge. Histamine (100 µM) induced a rapid but transient twofold increase in G' and G'' with maximum values 60 s after challenge. Posttreatment with cytochalasin D abolished thrombin-induced cell stiffening. G' increased with frequency following a power law with exponent 0.214. G'' increased proportionally to G' up to 10 Hz but showed a steeper rise at higher frequencies. Thrombin caused a fall in the power-law exponent (0.164). In measurements with acetylated low-density lipoprotein-coated beads, minor changes (<20%) were observed in G' and G'' after the addition of thrombin and histamine. F-actin staining revealed that thrombin and histamine induced a profound reorganization of the actin cytoskeleton at the cell periphery and formation of actin bundles. In the mechanically dynamic environment of the lung, cell stiffening induced by thrombin and histamine increases centripetal tension, which could contribute to alveolar barrier dysfunction.

cell mechanics; cell rheology; cytoskeleton mechanics; magnetic twisting cytometry; cytoskeleton reorganization

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