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Am J Physiol Cell Physiol (February 22, 2006). doi:10.1152/ajpcell.00411.2005
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Submitted on August 12, 2005
Accepted on February 19, 2006

The role of boundary conditions in the experimental model of wound healing

Djordje L Nikolic1, Alistair N Boettiger2, Dafna Bar-Sagi3, Jeffrey D Carbeck1, and Stanislav Y Shvartsman4*

1 Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA
2 Physics, Princeton University, Princeton, NJ, USA
3 Chemical Engineering, Princeton University, Princeton, NJ, USA
4 Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA; Department of Molecular Genetics and Microbiology, SUNY Stony Brook, Stony Brook, NY, none

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

Coordinated cell movements in epithelial layers are essential for proper tissue morphogenesis and homeostasis, but our understanding of the mechanisms that coordinate the behavior of multiple cells in these processes is far from complete. Recent experiments with MDCK epithelial monolayers revealed a wave-like pattern of injury-induced MAPK activation and showed that it is essential for collective cell migration after wounding. To investigate the effects of the different aspects of wounding on cell sheet migration, we engineered a system that allowed us to dissect the classic wound healing assay. We studied MDCK sheet migration under 3 different conditions: (1) the classic wound healing assay, (2) empty space induction, where a confluent monolayer is grown adjacent to a slab of polydimethylsiloxane (PDMS) and the monolayer is not injured but allowed to migrate upon removal of the slab, and (3) injury via PDMS membrane peel-off, where an injured monolayer migrates on plain tissue culture surface as in the case of empty space induction allowing for direct comparison. By tracking the motion of individual cells within the sheet under these three conditions, we show how the dynamics of the individual cells' motion is responsible for the coordinated migration of the sheet and is coordinated with the activation of ERK 1/2 MAPK. In addition, we demonstrate that the propagation of the waves of MAPK activation depends on the generation of reactive oxygen species at the wound edge.




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