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Biophys. J. BioFAST: First Published August 4, 2006. doi:10.1529/biophysj.106.084988
© 2006 by the Biophysical Society.

A more recent version of this article appeared on November 1, 2006.
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CELL BIOPHYSICS

Microrheology and ROCK signaling of human endothelial cells embedded in a 3D matrix

Porntula Panorchan 1, Jerry S.H. Lee 1, Thomas P. Kole 1, Yiider Tseng 1 and Denis Wirtz 1*

1 Johns Hopkins University

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

Submitted on March 10, 2006
Revised on May 8, 2006
Accepted on 18 July 2006


  Abstract
Cell function is profoundly affected by the geometry of the extracellular environment confining the cell. Whether and how cells plated on a two-dimensional matrix or embedded in a three-dimensional (3D) matrix mechanically sense the dimensionality of their environment is mostly unknown, partly because individual cells in an extended matrix are inaccessible to conventional cell-mechanics probes. Here we develop a functional assay based on multiple particle tracking microrheology coupled with ballistic injection of nanoparticles to measure the local intracellular micromechanical properties of individual cells embedded inside a matrix. As a proof of principle of this assay, we probe the mechanical properties of the cytoplasm of individual human umbilical vein endothelial cells (HUVECs) embedded in a 3D peptide hydrogel in the presence/absence of vascular endothelial growth factor (VEGF). We found that VEGF treatment, which enhances endothelial migration, increases the compliance and reduces the elasticity of the cytoplasm of HUVECs in a matrix. This VEGF-induced softening response of the cytoplasm is abrogated by specific ROCK inhibition. These results establish combined particle-tracking microrheology and ballistic injection as the first method able to probe the micromechanical properties and mechanical response to agonists and/or drug treatments of individual cells inside a matrix. These results suggest that ROCK plays an essential role in the regulation of the intracellular mechanical response to VEGF of endothelial cells in a 3D matrix.

Key Words: 3D matrix, Cell mechanics, Rho GTPases, endothelial cells




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Copyright © 2006 by the Biophysical Society.