title: eNOS transfection of adipose-derived stem cells yields bioactive nitric oxide production and improved results in vascular tissue engineering.
abstract: This study evaluates the durability of a novel tissue engineered blood vessel (TEBV) created by seeding a natural vascular tissue scaffold (decellularized human saphenous vein allograft) with autologous adipose-derived stem cells (ASC) differentiated into endothelial-like cells. Previous work with this model revealed the graft to be thrombogenic, likely due to inadequate endothelial differentiation as evidenced by minimal production of nitric oxide (NO). To evaluate the importance of NO expression by the seeded cells, we created TEBV using autologous ASC transfected with the endothelial nitric oxide synthase (eNOS) gene to produce NO. We found that transfected ASC produced NO at levels similar to endothelial cell (EC) controls in vitro which was capable of causing vasorelaxation of aortic specimens ex vivo. TEBV (n = 5) created with NO-producing ASC and implanted as interposition grafts within the aorta of rabbits remained patent for two months and demonstrated a non-thrombogenic surface compared to unseeded controls (n = 5). Despite the xenograft nature of the scaffold, the TEBV structure remained well preserved in seeded grafts. In sum, this study demonstrates that upregulation of NO expression within adult stem cells differentiated towards an endothelial-like lineage imparts a non-thrombogenic phenotype and highlights the importance of NO production by cells to be used as endothelial cell substitutes in vascular tissue engineering applications.
authors: Department of Surgery, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Stephen
authors: S
authors: McIlhenny
authors: Department of Surgery, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Ping
authors: P
authors: Zhang
authors: Department of Surgery, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Thomas
authors: T
authors: Tulenko
authors: Department of Surgery, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Jason
authors: J
authors: Comeau
authors: Department of Surgery, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Sarah
authors: S
authors: Fernandez
authors: Department of Surgery, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Aleksandra
authors: A
authors: Policha
authors: Department of Surgery, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Matthew
authors: M
authors: Ferroni
authors: Department of Orthopaedic Research, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Elizabeth
authors: E
authors: Faul
authors: Department of Surgery, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Gabor
authors: G
authors: Bagameri
authors: Department of Orthopaedic Research, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Irving
authors: I
authors: Shapiro
authors: Department of Surgery, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Paul
authors: P
authors: DiMuzio
mesh_terms: D017667:Adipocytes / Q000166:cytology*; D000273:Adipose Tissue / Q000166:cytology; D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology*; D000818:Animals; D001011:Aorta / Q000473:pathology; D001012:Aorta, Abdominal / Q000473:pathology; D001681:Biological Assay; D002454:Cell Differentiation; D042783:Endothelial Cells / Q000166:cytology; D005822:Genetic Vectors; D064593:Heterografts; D006801:Humans; D018613:Microscopy, Confocal; D009569:Nitric Oxide / Q000737:chemistry*; D052250:Nitric Oxide Synthase Type III / Q000737:chemistry*; D010641:Phenotype; D011817:Rabbits; D051381:Rats; D013927:Thrombosis; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds; D015854:Up-Regulation
chemical_list: D009569:Nitric Oxide; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III
keywords: adult stem cells; blood vessel prosthesis; differentiation; eNOS transfection; endothelium; nitric oxide synthase