Influence of cytoskeletal structure and mechanics on epithelial cell injury during cyclic airway reopening.

Although patients with acute respiratory distress syndrome require mechanical ventilation, these ventilators often exacerbate the existing lung injury. For example, the cyclic closure and reopening of fluid-filled airways during ventilation can cause epithelial cell (EpC) necrosis and barrier disruption. Although much work has focused on minimizing the injurious mechanical forces generated during ventilation, an alternative approach is to make the EpC less susceptible to injury by altering the cell's intrinsic biomechanical/biostructural properties. In this study, we hypothesized that alterations in cytoskeletal structure and mechanics can be used to reduce the cell's susceptibility to injury during airway reopening. EpC were treated with jasplakinolide to stabilize actin filaments or latrunculin A to depolymerize actin and then exposed to cyclic airway reopening conditions at room temperature using a previously developed in vitro cell culture model. Actin stabilization did not affect cell viability but significantly improved cell adhesion primarily due to the development of more numerous focal adhesions. Surprisingly, actin depolymerization significantly improved both cell viability and cell adhesion but weakened focal adhesions. Optical tweezer based measurements of the EpC's micromechanical properties indicate that although latrunculin-treated cells are softer, they also have increased viscous damping properties. To further investigate the effect of "fluidization" on cell injury, experiments were also conducted at 37 degrees C. Although cells held at 37 degrees C exhibited no changes in cytoskeletal structure, they did exhibit increased viscous damping properties and improved cell viability. We conclude that fluidization of the actin cytoskeleton makes the EpC less susceptible to the injurious mechanical forces generated during cyclic airway reopening.

Viscoelasticity of aqueous telechelic poly(ethylene oxide) solutions: relaxation and structure.

We present a rheology study of associating polymers. The associating polymers are telechelic, composed of a water-soluble backbone (polyethylene oxide) terminated by hydrophobic moieties (C16H33). In aqueous solutions, these polymers self-assemble to form micellar structures. Above a critical concentration, approximately 1 wt % of polymer, bridging between the micelles forms a transient network. Traditionally, the viscoelastic response of these polymeric solutions has been described using the Maxwell model. In this work we measure the viscoelastic properties over an extended frequency range (0.01-6000 Hz) using microrheology, and show that at high frequencies the rheology behaves as the square root of the oscillation frequency. To fit the data, we use a combination of the Maxwell model and the Rouse model. The Maxwell model accounts for the hydrophobic associations between the polymeric micelles, and the Rouse model accounts for the microscopic dynamics of the individual micelles.

Correlated motions of two hydrodynamically coupled particles confined in separate quadratic potential wells.

We report a study of the correlated motions of two hydrodynamically coupled colloidal particles, each of which is trapped in a quadratic potential well defined by optical tweezers (optical traps). By setting one of the trapped particles into forced oscillation using oscillating optical tweezers, we measure the displacement and phase shift of each of the particles over a wide frequency range. From the in-phase and out-of-phase motions of both of the particles in the traps, we determine the correlated motions of the coupled mechanical system as a function of frequency. A theoretical model is developed to calculate the response tensor of the coupled mechanical system. The experimental results are in agreement with the prediction of the theoretical model. This method may be extended to more general applications, such as the investigation of the micromechanical properties of viscoelastic and/or heterogeneous media.