Proliferation of endothelial cells on surface-immobilized albumin-heparin conjugate loaded with basic fibroblast growth factor.
Seeding of endothelial cells (ECs) on the luminal surface of small-diameter vascular grafts is a promising method to avoid occlusion of these prostheses. Immobilization of basic fibroblast growth factor (bFGF) to substrates used to coat or fill porous prostheses may enhance the formation of a confluent monolayer of ECs. Human umbilical vein endothelial cells (HUVECs) were grown on bFGF-loaded albumin-heparin conjugate bound to CO2 gas-plasma-treated polystyrene. In the order of 2-3 ng/cm2 bFGF had to be immobilized to form a confluent monolayer of HUVECs. The most prominent effect of surface-immobilized bFGF was stimulation of the proliferation shortly after seeding, resulting within 3 days in confluent cell monolayers with high density. In contrast, in cultures with 0.3 ng/mL bFGF in the medium instead of bFGF bound to the surface, it took almost a week before the cell layers reached confluency. Binding of bFGF to heparin and the biological activity of bFGF towards ECs were not influenced by the (radio-)labeling of bFGF with iodine. However, only a minor part of the bFGF used in this study displayed heparin affinity. Furthermore, degradation and multimerization of labeled bFGF in time occurred when the growth factor was stored at 20 degrees -37 degrees C. This limits the use of labeled bFGF to short-term (hours) experiments. In conclusion, bFGF loading of vascular graft surfaces through complexation of bFGF with a heparin-containing matrix probably will lead to more rapid formation of a confluent monolayer of ECs on graft surfaces upon seeding of the cells.
Bos
G W
GW
Faculty of Chemical Technology, Section of Polymer Chemistry and Biomaterials and Institute for Biomedical Technology, University of Twente, Enschede, The Netherlands.
Scharenborg
N M
NM
Poot
A A
AA
Engbers
G H
GH
Beugeling
T
T
van Aken
W G
WG
Feijen
J
J
D001807:Blood Vessel Prosthesis; D002245:Carbon Dioxide; D018929:Cell Culture Techniques / Q000295:instrumentation; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005353:Fibronectins / Q000737:chemistry / Q000302:isolation & purification; D006493:Heparin / Q000737:chemistry*; D006801:Humans; D007457:Iodine Radioisotopes / Q000032:analysis; D008863:Microspheres; D009842:Oligopeptides / Q000502:physiology; D011137:Polystyrenes; D017469:Receptors, Fibronectin / Q000502:physiology; D019030:Receptors, Vitronectin / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012685:Sepharose; D012709:Serum Albumin / Q000737:chemistry*; D013242:Sterilization; D013462:Sulfur Radioisotopes / Q000032:analysis
D003470:Culture Media; D005353:Fibronectins; D007457:Iodine Radioisotopes; D009842:Oligopeptides; D011137:Polystyrenes; D017469:Receptors, Fibronectin; D019030:Receptors, Vitronectin; D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; D013462:Sulfur Radioisotopes; D016222:Fibroblast Growth Factor 2; D002245:Carbon Dioxide; C047981:arginyl-glycyl-aspartic acid; D006493:Heparin; D012685:Sepharose