High-affinity augmentation of endothelial cell attachment: long-term effects on focal contact and actin filament formation.

Coadsorption of high-affinity avidin with lower affinity cell adhesion protein fibronectin has been shown to significantly augment short-term (1 h) adhesion and spreading of endothelial cells; however, the longer term persistence of avidin binding and its effect on endothelial cell adhesion have not been addressed. In this study, the presence of avidin-biotin bonds 24 h after cell adhesion to the dual ligand surfaces was verified by laser confocal microscopy of a fluorescent avidin analog, streptavidin. Total internal reflection microscopy showed that the focal contact area, focal contact density, and cell spreading all increased significantly at 24 h compared to fibronectin-treated control surfaces. Focal contact area was identical when measured with cells that were labeled with either the fluorescent streptavidin or a carbocyanine dye incorporated in the cell membrane. Confocal images of stress fibers formed in cells adherent to dual ligand surfaces after 24 h were thicker and more numerous compared to cells adherent to fibronectin controls. The results indicate that 24 h after initial attachment avidin-biotin is localized to focal contacts on the basal surface and affects cell spreading, actin filament organization, and focal contact density.

Duke Scholars

Author George A. Truskey Biomedical Engineering

Author William M. Reichert Biomedical Engineering