Overexpression of eNOS decrease tissue factor (TF) level in CD34+ cells exhibit increased antithrombogenic property in small caliber vascular graft.
BACKGROUND
Endothelial progenitor cells (EPCs) have reduced expression of eNOS, this may decrease their antithrombogenic property when used as seeding cells for small caliber vascular graft. The aim of this study is to investigate whether overexpression of eNOS in EPCs can increase its antithrombogenic property and regulate tissue factor (TF) level.


METHODS
CD34+ cells were isolated from canine bone marrow. Differentiation of CD34 cells into endothelial cells was inducted by VEGF. Overexpression of eNOS in CD34+ cells were obtained by transfection with eNOS plasmid. TF expression was examined by western blot after TNFα stimulation. Platelets adhesion assay was performed to determine antiplatelet adhesion property of the cells in vitro. The cells were seeded onto the lumimal surface of small caliber vascular graft and implanted in vivo. The thrombopoiesis in vivo were examined by SEM.


RESULTS
Transfection with eNOS gene decreased the level of TF in CD34+ cells. The expression of TF increased after stimulation with TNFα in time dependent manner, this effect was abrogated by eNOS gene transfection. Overexpression of eNOS significantly inhibited the platelet adhesion on EPCs in vitro. Over expression of eNOS in CD34+ cells also decreased thrombopoiesis and fibrin adhesion onto the lumimal surface of small caliber vascular graft in vivo.


CONCLUSIONS
Overexpression of eNOS decrease TF level in CD34+ cells, and increase antithrombogenic property of small caliber vascular graft.
Department of Cardiothoracic Surgery, Shanghai Ninth People' s Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, China.
Feng
F
Wang
Department of Cardiothoracic Surgery, Affiliated Hospital of Jiansu University, Jiansu, China.
Gang
G
Li
Department of Cardiothoracic Surgery, Shanghai Ninth People' s Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, China.
Xin
X
Guan
Department of Respiratory Medicine, Shanghai Ninth People's Hospital Affiliated to Shanghai Jiao Tong, University School of Medicine, Shanghai, China.
Zhaoqing
Z
Han
Department of Orthopedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China.
Xiaowei
X
Yu
Department of Cardiothoracic Surgery, Affiliated Hospital of Jiangnan University, Wuxi, China - wxsytsg@126.com.
Qingjun
Q
You
D000818:Animals; D018952:Antigens, CD34; D001672:Biocompatible Materials; D001807:Blood Vessel Prosthesis; D002478:Cells, Cultured; D004285:Dogs; D066026:Endothelial Progenitor Cells / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D013925:Thromboplastin / Q000378:metabolism*; D034061:Thrombopoiesis / Q000502:physiology*; D013927:Thrombosis; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D015854:Up-Regulation; D042442:Vascular Endothelial Growth Factors
D018952:Antigens, CD34; D001672:Biocompatible Materials; D014409:Tumor Necrosis Factor-alpha; D042442:Vascular Endothelial Growth Factors; D013925:Thromboplastin; D052250:Nitric Oxide Synthase Type III