High-affinity augmentation of endothelial cell attachment: Long-term effects on focal contact and actin filament formation
✍ Scribed by Mathur, Anshu B. ;Chan, Bernard P. ;Truskey, George A. ;Reichert, William M.
- Book ID
- 102293433
- Publisher
- John Wiley and Sons
- Year
- 2003
- Tongue
- English
- Weight
- 357 KB
- Volume
- 66A
- Category
- Article
- ISSN
- 0021-9304
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✦ Synopsis
Abstract
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. © 2003 Wiley Periodicals, Inc. J Biomed Mater Res 66A: 729–737, 2003