Submitted on November 16, 2007
Accepted on May 8, 2008
Localized elasticity measured in epithelial cells migrating at a wound edge using atomic force microscopy
Ajay A. Wagh1, Esra Roan2, Kenneth E. Chapman1, Leena P. Desai1, David A. Rendon3, Eugene C. Eckstein2, and Christopher M. Waters4*
1 Physiology, University of Tennessee Health Science Center, Memphis, Tennessee, United States
2 Biomedical Engineering, University of Memphis, Memphis, Tennessee, United States
3 Biomedical Engineering, University of Tennessee Health Science Center, Memphis, Tennessee, United States
4 Physiology, University of Tennessee Health Science Center, Memphis, Tennessee, United States; Biomedical Engineering, University of Tennessee Health Science Center, Memphis, Tennessee, United States
* To whom correspondence should be addressed. E-mail: cwaters2{at}utmem.edu.
Restoration of lung homeostasis following injury requires efficient wound healing by the epithelium. The mechanisms of lung epithelial wound healing include cell spreading and migration into the wounded area and later cell proliferation. We hypothesized that mechanical properties of cells vary near the wound edge and this may provide cues to direct cell migration. To investigate this hypothesis, we measured variations in the stiffness of migrating human bronchial epithelial cells (16HBE cells) approximately two hours after applying a scratch wound. We used atomic force microscopy (AFM) in contact mode to measure the cell stiffness in 1.5 um square regions at different locations relative to the wound edge. In regions far from the wound edge (>2.75 mm) there was substantial variation in the elastic modulus in specific cellular regions, but the median values measured from multiple fields were consistently lower than 5 kPa. At the wound edge cell stiffness was significantly lower within the first 5 um, but increased significantly between 10 and 15 um before decreasing again below the median values away from the wound edge. When cells were infected with an adenovirus expressing a dominant negative form of RhoA, cell stiffness was significantly decreased compared with cells infected with a control adenovirus. In addition expression of dominant negative RhoA abrogated the peak increase in stiffness near the wound edge. These results suggest that cells near the wound edge undergo localized changes in cellular stiffness that may provide signals for cell spreading and migration.
