Time-course for in vitro development of basement membrane, gap junctions, and repair by adult endothelial cells seeded on precoated ePTFE.
OBJECTIVE
To establish some functional characteristics after in vitro endothelialisation of precoated ePTFE grafts.


METHODS
Saphenous vein endothelial cells were confluently seeded on human serum-precoated ePTFE. Collagen I-precoated ePTFE served as control. Time-courses for development of laminin, collagen IV and connexin43 was followed-up with immunofluorescence. Endothelial migration and proliferation was studied with cresyl violet staining and tritiated thymidine-labelling.


RESULTS
Six hours post-seeding, basement membrane components were visualised subendothelially as thin fibrils. After 24 h, there was a distinct fibril network, which did not seem to be further enhanced. Connexin43 was detected 6 h post-seeding, and the number of intercellular connections appeared to be similar up to day 7. The time-courses were similar on serum- and collagen I-precoated ePTFE. Endothelialisation of a 4 mm wide gap showed migrating fronts largely composed of proliferating cells. The process was completed within 10 days on collagen I-precoated ePTFE, and 15 days on serum-precoated ePTFE.


CONCLUSIONS
These findings suggest human endothelial cells, seeded on precoated ePTFE, maintain some differentiated functions seen in vivo. This is of importance for cell retention, functionality and endothelial repair of in vitro endothelialised ePTFE-grafts after implantation in man.
Jansson
K
K
Karolinska Institutet, Department of Surgical Sciences, Karolinska Hospital, Stockholm, Sweden.
Bengtsson
L
L
Haegerstrand
A
A
D001485:Basement Membrane / Q000254:growth & development; D001807:Blood Vessel Prosthesis; D002455:Cell Division; D002465:Cell Movement; D020099:Coated Materials, Biocompatible; D017630:Connexins / Q000096:biosynthesis; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology* / Q000637:transplantation; D017629:Gap Junctions / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D011138:Polytetrafluoroethylene; D012501:Saphenous Vein / Q000166:cytology; D013997:Time Factors
D020099:Coated Materials, Biocompatible; D017630:Connexins; D011138:Polytetrafluoroethylene