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

被引:19
作者
Mathur, AB
Chan, BP
Truskey, GA
Reichert, WM
机构
[1] Duke Univ, Ctr Cellular & Biosurface Engn, Durham, NC 27708 USA
[2] Duke Univ, Dept Biomed Engn, Durham, NC 27708 USA
来源
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A | 2003年 / 66A卷 / 04期
关键词
cell adhesion; focal contacts; cytoskeleton; stress fiber; TIRFM; fibronectin; integrins; avidin; biotin;
D O I
10.1002/jbm.a.10581
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
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. (C) 2003 Wiley Periodicals, Inc.
引用
收藏
页码:729 / 737
页数:9
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