title: Extracellular matrix preparation of expanded polytetrafluoroethylene grafts seeded with endothelial cells: influence on early platelet deposition, cellular growth, and luminal prostacyclin release.
abstract: The importance of blood and extracellular matrix precoating of expanded polytetrafluoroethylene (ePTFE) grafts on the effectiveness of endothelial cell (EC) seeding was assessed in a canine experimental model. Part I of the study documented ex vivo platelet deposition in 256 ePTFE grafts, 6 cm x 4 mm internal diameter, after implantation as femoral artery-femoral vein or carotid artery-jugular vein arteriovenous shunts. These conduits were precoated with blood, fibronectin, laminin, or collagen type IV with laminin, after which they were seeded with enzymatically derived and cultivated venous canine endothelium at a density of 30,000 to 40,000 EC/cm2 of graft surface. Luminal deposition of Indium 111-labeled platelets, expressed as 10(8) platelets/cm2, at 30 minutes (n = 176) and 24 hours (n = 80), respectively, was 2.29 and 0.30 for blood, 2.83 and 0.37 for fibronectin, 0.99 and 0.08 for laminin, and 0.98 and 0.11 for collagen type IV with laminin. Part II of the study documented in vivo luminal EC coverage at 14 days of 6 cm x 4 mm internal diameter ePTFE femoral or carotid arterial grafts (n = 8) prepared in the same manner as part I ex vivo shunt grafts. EC coverage with blood, fibronectin, laminin, and collagen type IV with laminin preparation was 42%, 49%, 44%, and 52%, respectively. The graft:carotid artery ratio of luminal 6-keto-PGF1 alpha release at 14 days with these same four preparations was 0.38, 0.31, 0.35, and 0.32, respectively. Precoatings of ePTFE prostheses with fibronectin, laminin, and collagen type IV are known to enhance the initial attachment of seeded EC. Fibronectin caused an insignificant increase in early platelet accumulation; laminin or laminin with collagen type IV preparations were associated with significantly less (p less than 0.005) deposition of platelets when compared to whole blood preparations. Most importantly, none of the four preparation techniques resulted in different in vivo rates of EC growth or luminal release of prostacyclin from conduits studied 14 days after implantation.
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor.
authors.forename: G B
authors.initials: GB
authors.lastname: Köveker
authors.forename: L M
authors.initials: LM
authors.lastname: Graham
authors.forename: W E
authors.initials: WE
authors.lastname: Burkel
authors.forename: R
authors.initials: R
authors.lastname: Sell
authors.forename: T W
authors.initials: TW
authors.lastname: Wakefield
authors.forename: K
authors.initials: K
authors.lastname: Dietrich
authors.forename: J C
authors.initials: JC
authors.lastname: Stanley
mesh_terms: D000818:Animals; D001166:Arteriovenous Shunt, Surgical; D001792:Blood Platelets / Q000502:physiology*; D002339:Carotid Arteries / Q000601:surgery*; D002455:Cell Division; D003094:Collagen; D004285:Dogs; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology / Q000637:transplantation*; D011464:Epoprostenol / Q000378:metabolism*; D005109:Extracellular Matrix / Q000502:physiology*; D005260:Female; D005263:Femoral Artery / Q000601:surgery*; D005353:Fibronectins; D006801:Humans; D007797:Laminin; D051379:Mice; D011138:Polytetrafluoroethylene; D014656:Vascular Surgical Procedures / Q000379:methods*
chemical_list: D005353:Fibronectins; D007797:Laminin; D011138:Polytetrafluoroethylene; D003094:Collagen; D011464:Epoprostenol