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Am J Physiol Cell Physiol (August 14, 2002). doi:10.1152/ajpcell.00341.2002
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Articles in PresS, published online ahead of print August 14, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00341.2002
Submitted on July 23, 2002
Accepted on August 6, 2002

Measurement of stretch-induced loss of alveolar epithelial barrier integrity using a novel in vitro method

Kenneth J Cavanaugh, Jr.1 and Susan S Margulies1*

1 Bioengineering, University of Pennsylvania, Philadelphia, PA, USA

* To whom correspondence should be addressed. E-mail: margulie{at}seas.upenn.edu.

Mechanical ventilation with high tidal volumes has been shown to contribute to the formation or worsening of interstitial and alveolar edema. Previously we have shown that application of large biaxial deformations in vitro perturbs the concentration and distribution of functional tight junction proteins in alveolar epithelial cells. Using a novel method, we determined that applied epithelial strain increases paracellular permeability in a dose- and rate-dependent manner. Primary rat alveolar epithelial cells were subjected to a 12%, 25%, or 37% change in surface area ({Delta}SA) cyclic equibiaxial stretch for 1 hr. Cells were also stretched non-cyclically at 25% {Delta}SA for 1 hr. During the experimental period, a fluorescently-tagged ouabain derivative was added to the apical fluid. Evidence of binding indicated functional failure of the paracellular transport barrier. Percent of field area stained was quantified from microscopic images. There was no significant evidence of basolateral fluorescent staining at 12% {Delta}SA or at 25% {Delta}SA applied cyclically or statically. However, cyclic stretch at 37% {Delta}SA resulted in significantly more staining than in unstretched cells (p<0.0001), or those stretched at either 12% (p<0.0001) or 25% cyclic (p<0.0005) or static (p<0.05) {Delta}SA. These results suggest that large cyclic tidal volumes may increase paracellular permeability, potentially resulting in alveolar flooding.




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