Journal of Applied Physiology AJP: Renal Physiology
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J Appl Physiol (June 5, 2008). doi:10.1152/japplphysiol.00958.2007
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Submitted on September 11, 2007
Accepted on May 29, 2008

Atomic force microscope elastography reveals phenotypic differences in alveolar cell stiffness

Evren U. Azeloglu1, Jahar Bhattacharya2, and Kevin D. Costa3*

1 Biomedical Engineering, Columbia University, New York, New York, United States; , United States
2 Medicine, St. Luke's-Roosevelt Hospital, New York, New York, United States
3 Biomedical Engineering, Columbia University, New York, New York, United States

* To whom correspondence should be addressed. E-mail: kdc17{at}columbia.edu.

To understand the connection between alveolar mechanics and key biochemical events such as surfactant secretion, one first needs to characterize the underlying mechanical properties of the lung parenchyma and its cellular constituents. In this study, the mechanics of three major cell types from the neonatal rat lung were studied; primary alveolar type I (AT1) and type II (AT2) epithelial cells, and lung fibroblasts were isolated using enzymatic digestion. Atomic force microscopy (AFM) indentation was used to map the 3-D distribution of apparent depth-dependent pointwise elastic modulus. Histograms of apparent modulus data from all three cell types indicated non-Gaussian distributions that were highly skewed, and appeared multi-modal for AT2 cells and fibroblasts. Nuclear stiffness in all three cell types were similar (2.5±1.0 kPa in AT1 vs. 3.1±1.5 kPa in AT2 vs. 3.3±0.8 kPa in fibroblasts; n = 10 each) whereas cytoplasmic moduli were significantly higher in fibroblasts and AT2 cells (6.0±2.3 kPa and 4.7±2.9 kPa vs. 2.5±1.2 kPa). In both epithelial cell types f-actin was arranged in sparse clusters, whereas prominent actin stress fibers were observed in lung fibroblasts. No systematic difference in actin or microtubule organization was noted between AT1 and AT2 cells. AFM elastography combined with live-cell fluorescence imaging revealed that the stiffer measurements in AT2 cells often colocalized with lamellar bodies. These findings partially explain reported heterogeneity of alveolar cell deformation during in situ lung inflation, and provide needed data for better understanding how mechanical stretch influences surfactant release.




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