ORIGINAL ARTICLE  CARDIAC SECTION 

The Journal of Cardiovascular Surgery 2019 February;60(1):136-43

DOI: 10.23736/S0021-9509.18.08951-6

Copyright © 2015 EDIZIONI MINERVA MEDICA

language: English

Overexpression of eNOS decrease tissue factor (TF) level in CD34+ cells exhibit increased antithrombogenic property in small caliber vascular graft

Feng WANG ¹, Gang LI ², Xin GUAN ¹, Zhaoqing HAN ³, Xiaowei YU ⁴, Qingjun YOU ⁵ ✉

¹ Department of Cardiothoracic Surgery, Shanghai Ninth People’ s Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, China; ² Department of Cardiothoracic Surgery, Affiliated Hospital of Jiansu University, Jiansu, China; ³ Department of Respiratory Medicine, Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiao Tong, University School of Medicine, Shanghai, China; ⁴ Department of Orthopedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai, China; ⁵ Department of Cardiothoracic Surgery, Affiliated Hospital of Jiangnan University, Wuxi, China

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.

KEY WORDS: Vascular grafting - Endothelial progenitor cells - Thromboplastin