title: eNOS-overexpressing endothelial cells inhibit platelet aggregation and smooth muscle cell proliferation in vitro.
abstract: Endothelial cell seeding of synthetic small diameter vascular grafts (SSDVG) has been shown to diminish thrombosis and intimal hyperplasia, resulting in improved graft patency. However, endothelial cell retention on seeded grafts when exposed to physiological shearing conditions remains poor. We report that the genetic engineering of endothelial cells to overexpress endothelial nitric oxide synthase (eNOS), may create improved anti-thrombotic and anti-hyperplastic endothelial cell phenotypes for SSDVG seeding. eNOS-overexpressing endothelial cells may potentially overcome the biochemical loss due to shear induced reduction in endothelial cell coverage on SSDVG. Bovine aortic endothelial cells (BAEC) were transfected with the human eNOS gene, and co-incubated with either human whole blood or bovine aortic smooth muscle cells (BASMC) in vitro. eNOS-transfected BAEC significantly overexpressed eNOS compared to control beta-Gal-transfected and untransfected BAEC up to 120 h post transfection. In co-incubation and co-culture assays, human platelet aggregation decreased by 46% and BASMC proliferation decreased by 67.2% when compared to incubation with untransfected BAEC.
authors.affiliation: Biomaterials, Cell and Tissue Engineering Laboratory, Department of Biomedical Engineering, Case Western Reserve University, Cleveland, Ohio 44106-7207, USA.
authors.forename: K N
authors.initials: KN
authors.lastname: Kader
authors.forename: R
authors.initials: R
authors.lastname: Akella
authors.forename: N P
authors.initials: NP
authors.lastname: Ziats
authors.forename: L A
authors.initials: LA
authors.lastname: Lakey
authors.forename: H
authors.initials: H
authors.lastname: Harasaki
authors.forename: J P
authors.initials: JP
authors.lastname: Ranieri
authors.forename: R V
authors.initials: RV
authors.lastname: Bellamkonda
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000502:physiology*; D002417:Cattle; D002450:Cell Communication; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D005818:Genetic Engineering; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000502:physiology*; D019001:Nitric Oxide Synthase / Q000502:physiology*; D052250:Nitric Oxide Synthase Type III; D010974:Platelet Aggregation
chemical_list: C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III