Improving endothelial cell retention for single stage seeding of prosthetic grafts: use of polymer sequences of arginine-glycine-aspartate.
OBJECTIVE
single stage seeding within the timeframe of a typical vascular operation has not been successful. One reason for this is poor cell adherence to the graft lumen once exposed to pulsatile blood flow. In this study we have carried out investigations with the use of two different fibronectin-based peptides, fibronectin-like engineered protein polymer (FEPP) which contains multiple copies of arginine-glycine-aspartate (RGD) and fibronectin adhesion promoting peptide (FAPP) to improve cell adherence.


METHODS
FAPP and FEPP were coated onto native polyurethane and heparinised polyurethane grafts. The grafts were then seeded for either 1 or 2h with human umbilical vein endothelial cells (HUVEC). After the incubation period the cells were washed off and cell retention was calculated. Cell metabolism was measured using Alamar Blue, and confirmed with scanning electron microscopy (SEM).


RESULTS
heparinised grafts coated with FEPP showed the best cell retention after both 1 and 2h seeding (80+/-4% vs 81+/-3%). This graft had no significant difference in cell retention after both times whilst all the other grafts had better cell retention after a 2h seeding. The Alamar blue and SEM results confirmed cell viability and function for all graft types.


CONCLUSIONS
heparinised graft coated with FEPP allows significant cell retention after only 1h of seeding and shows promise for single stage seeding.
Tiwari
A
A
Tissue Engineering Centre, University Department of Surgery, Royal Free and University College Medical School, University College London and The Royal Free Hospital, London, UK.
Kidane
A
A
Salacinski
H
H
Punshon
G
G
Hamilton
G
G
Seifalian
A M
AM
D001807:Blood Vessel Prosthesis; D002448:Cell Adhesion / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000473:pathology* / Q000648:ultrastructure; D005353:Fibronectins / Q000494:pharmacology*; D006801:Humans; D009842:Oligopeptides / Q000494:pharmacology*; D011140:Polyurethanes / Q000494:pharmacology*; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*; D014471:Umbilical Veins / Q000187:drug effects* / Q000473:pathology* / Q000648:ultrastructure
D005353:Fibronectins; D009842:Oligopeptides; D011140:Polyurethanes; C047981:arginyl-glycyl-aspartic acid