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Am J Physiol Cell Physiol 291: C718-C725, 2006. First published August 23, 2006; doi:10.1152/ajpcell.00127.2005
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PROTEIN AND VESICLE TRAFFICKING, CYTOSKELETON

Chondrocyte intracellular calcium, cytoskeletal organization, and gene expression responses to dynamic osmotic loading

Pen-hsiu Grace Chao,1,2 Alan C. West,2 and Clark T. Hung1

1Department of Biomedical Engineering and 2Department of Chemical Engineering, Columbia University, New York, New York

Submitted 17 March 2005 ; accepted in final form 24 April 2006

While chondrocytes in articular cartilage experience dynamic stimuli from joint loading activities, few studies have examined the effects of dynamic osmotic loading on their signaling and biosynthetic activities. We hypothesize that dynamic osmotic loading modulates chondrocyte signaling and gene expression differently than static osmotic loading. With the use of a novel microfluidic device developed in our laboratory, dynamic hypotonic loading (–200 mosM) was applied up to 0.1 Hz and chondrocyte calcium signaling, cytoskeleton organization, and gene expression responses were examined. Chondrocytes exhibited decreasing volume and calcium responses with increasing loading frequency. Phalloidin staining showed osmotic loading-induced changes to the actin cytoskeleton in chondrocytes. Real-time PCR analysis revealed a stimulatory effect of dynamic osmotic loading compared with static osmotic loading. These studies illustrate the utility of the microfluidic device in cell signaling investigations, and their potential role in helping to elucidate mechanisms that mediate chondrocyte mechanotransduction to dynamic stimuli.

cartilage; calcium signaling; actin cytoskeleton; aggrecan


Address for reprint requests and other correspondence: C. T. Hung, Dept. of Biomedical Engineering, 351 Engineering Terrace, 1210 Amsterdam Ave., MC 8904, New York, NY 10027 (e-mail: cth6{at}columbia.edu)






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