Effect of retroviral transduction on human endothelial cell phenotype and adhesion to Dacron vascular grafts.
OBJECTIVE
Retroviral transduction for genetic enhancement of endothelial cell (EC) anti-thrombotic phenotype offers potential for improving the clinical success of vascular graft seeding; however, application of this technique may bring concomitant alteration in cell functionality.


METHODS
Human microvascular ECs were transduced with a retroviral vector encoding for the marker gene beta-galactosidase. Transduced endothelial cells (rtECs) and nontransduced endothelial cells (ntECs) were evaluated by flow cytometry for expression of intercellular adhesion molecule (ICAM)-1 and tissue factor (TF) on both smooth (coverslips) and graft (Dacron, 6 mm inside diameter) surfaces under static and shear exposed conditions. Graft EC retention was measured after 6-hour pulsatile perfusions. Platelet and neutrophil adherence was measured on perfused coverslips.


RESULTS
Lower levels of ICAM-1 were expressed by rtECs on coverslips under both static (p < 0.01 vs static ntECs) and shear exposed conditions (p < 0.01 vs static and shear ntECs). Accordingly, fewer polymorphonuclear leukocytes adhered to rtEC monolayers (p < 0.01 vs ntECs). No difference in ICAM-1 and TF expression by static graft seeded rtECs and ntECs was observed. However, graft-seeded rtECs that were exposed to wall shear stress displayed less TF than sheared ntECs (p < 0.05). Transduction did not affect EC retention to the sheared graft surface.


CONCLUSIONS
These data suggest that retroviral transduction does not elicit a prothrombotic/proinflammatory phenotype, rather indices of these states appear in some conditions to be reduced. Further, transduction does not adversely affect EC adherence to Dacron graft surfaces under arterial hemodynamics.
Jankowski
R J
RJ
Bioengineering Program, University of Pittsburgh, Pa., USA.
Severyn
D A
DA
Vorp
D A
DA
Wagner
W R
WR
D001792:Blood Platelets / Q000502:physiology; D001807:Blood Vessel Prosthesis; D002448:Cell Adhesion; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism*; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D009504:Neutrophils / Q000502:physiology; D010641:Phenotype; D011093:Polyethylene Terephthalates; D012190:Retroviridae; D013925:Thromboplastin / Q000378:metabolism; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000235:genetics
D011093:Polyethylene Terephthalates; D018799:Intercellular Adhesion Molecule-1; D013925:Thromboplastin; D001616:beta-Galactosidase