Synergistic effect of high-affinity binding and flow preconditioning on endothelial cell adhesion.
The current study examined whether the combined introduction of high-affinity avidin-biotin bonds and fibronectin-integrin bonds (i.e., dual ligand treatment) would further augment the adhesion of flow-preconditioned endothelial cells to model substrates via contributions to the actin cytoskeleton and the formation of focal contacts. Human umbilical vein endothelial cells (HUVEC) were grown under static conditions or exposed to a flow-preconditioning regimen for 24 h. Cell retention was determined by exposure to 75 dynes/cm(2). The combination of flow preconditioning and the dual ligand treatment yielded higher cell retention under flow compared to the cells adherent via fibronectin-integrin bonds only. This increase in adhesion strength correlated with a greater focal contact area. Elongation of the HUVEC occurred after exposure to flow preconditioning; however, orientation of dual ligand adherent cells was restricted due to the presence of the high-affinity ligand. Flow-preconditioned cells showed increased stress fiber formation compared to nonconditioned cells although the stress fibers per cell for flow-preconditioned cells were the same on both the ligand systems employed. The results indicate that enhanced adhesion strength is due to a combination of increased focal contact area, stress fiber formation, and cell alignment.
Mathur
Anshu B
AB
Department of Biomedical Engineering, Duke University, Durham, North Carolina 27708-0281, USA.
Truskey
George A
GA
Reichert
William M
WM
D001360:Avidin; D001710:Biotin; D002448:Cell Adhesion; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology*; D005353:Fibronectins / Q000648:ultrastructure; D006801:Humans; D008024:Ligands; D018625:Microscopy, Atomic Force; D011485:Protein Binding
D005353:Fibronectins; D008024:Ligands; D001360:Avidin; D001710:Biotin