In vitro adherence and kinetics studies of adult human endothelial cell seeded polytetrafluoroethylene and gelatin impregnated Dacron grafts.
Lining the luminal surface of small diameter vascular prostheses with living endothelial cells reduces thrombogenicity, decreases infection and improves patency. In vitro adherence and kinetics studies of adult human endothelial cell seeded Polytetrafluoroethylene (ePTFE) and Gelatin impregnated Dacron (Gelseal) were performed. Endothelial cell adherence on ePTFE and Gelseal coated with collagen IV, fibronectin and preclot matrices was compared. Untreated ePTFE and Gelseal were also used. Ten graft segments in each group coated with these matrices were incubated with radio-labelled adult human endothelial cells for 30, 60 and 90 min. Labelled endothelial cells seeded in supra-confluent densities on fibronectin coated ePTFE and Gelseal grafts were used for kinetic studies. Resultant endothelial cell monolayers were then exposed to varying shear stress at flow rates of 200 and 300 ml/min in an artificial flow circuit. Endothelial cell attachment to untreated grafts was poor and a suitable matrix significantly improved adherence with fibronectin and preclot but less so with collagen. A 30 min incubation was sufficient for optimum cell attachment. Cell adherence to ePTFE was significantly better than Gelseal. Scanning electron micrographs (SEM) of ePTFE showed preferential attachment to the nodes whilst on Gelseal, cells conformed to Dacron fibres at different levels and directions. Rapidly formed endothelial cell monolayers on ePTFE and Gelseal grafts resisted shear stress of flow with significant cell retention at 2 h. There was patchy coverage of both grafts with evidence of bridging of gaps between individual fibres in Gelseal.
Vohra
R
R
Department of Vascular Surgery, Manchester Royal Infirmary, U.K.
Thomson
G J
GJ
Carr
H M
HM
Sharma
H
H
Welch
M
M
Walker
M G
MG
D001807:Blood Vessel Prosthesis; D002448:Cell Adhesion; D003094:Collagen; D004730:Endothelium, Vascular / Q000166:cytology*; D005353:Fibronectins; D005780:Gelatin; D006083:Graft Occlusion, Vascular / Q000517:prevention & control*; D006801:Humans; D066298:In Vitro Techniques; D008855:Microscopy, Electron, Scanning; D011093:Polyethylene Terephthalates; D011138:Polytetrafluoroethylene
D005353:Fibronectins; D011093:Polyethylene Terephthalates; D005780:Gelatin; D011138:Polytetrafluoroethylene; D003094:Collagen