Endothelial cell seeding of (heparinized) collagen matrices: effects of bFGF pre-loading on proliferation (after low density seeding) and pro-coagulant factors.
Endothelial cell seeding to improve the performance of small-diameter vascular grafts requires a suitable substrate, such as crosslinked collagen. In addition to providing a suitable substrate for adhesion and growth of endothelial cells, proliferation of seeded endothelial cells can be enhanced by local, sustained release of basic fibroblast growth factor (bFGF, a heparin-binding growth factor for endothelial cells). We have previously shown that collagen crosslinked using N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS) supports adhesion and proliferation of human umbilical vein endothelial cells (HUVECs). In the present study, HUVECs were seeded on (heparinized) EDC/NHS-crosslinked collagen, pre-loaded with bFGF. Proliferation of HUVECs on (heparinized) crosslinked collagen increased with increasing amounts of pre-loaded bFGF. The minimal cell-seeding density required for proliferation proved to be very low after pre-loading the substrates with bFGF, and was 4-fold lower for heparinized crosslinked collagen compared to crosslinked collagen (250 versus 1000 cells/cm(2)). Pro-coagulant properties (von Willebrand factor secretion and tissue factor expression) of HUVECs seeded on (heparinized) crosslinked collagen, with or without pre-loading of bFGF, were comparable to those of HUVECs on TCPS. It is concluded that heparinized, EDC/NHS-crosslinked collagen pre-loaded with bFGF is a candidate matrix for in vivo endothelial cell seeding of synthetic vascular graft materials.
Wissink
M J
MJ
Institute for Biomedical Technology, Polymer Chemistry and Biomaterials Group, Department of Chemical Technology, University of Twente, P.O. Box 217, 7500 AE, Enschede, The Netherlands.
Beernink
R
R
Scharenborg
N M
NM
Poot
A A
AA
Engbers
G H
GH
Beugeling
T
T
van Aken
W G
WG
Feijen
J
J
D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003029:Coagulants / Q000494:pharmacology*; D003094:Collagen / Q000737:chemistry*; D003432:Cross-Linking Reagents; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005353:Fibronectins / Q000096:biosynthesis; D006493:Heparin / Q000737:chemistry*; D006801:Humans; D014471:Umbilical Veins / Q000166:cytology
D003029:Coagulants; D003432:Cross-Linking Reagents; D005353:Fibronectins; D016222:Fibroblast Growth Factor 2; D006493:Heparin; D003094:Collagen