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Overexpression of eNOS decrease tissue factor (TF) level in CD34+ cells exhibit increased antithrombogenic property
Feng Wang1, Gang Li2, Xin Guan1
1Department of Cardiothoracic Surgery, Shanghai Ninth People' s Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, 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.
Insights
Overexpressing endothelial nitric oxide synthase (eNOS) in endothelial progenitor cells (EPCs) reduces tissue factor (TF) and enhances antithrombogenic properties for vascular grafts.
Area of Science:
- Biomedical Engineering
- Vascular Biology
- Cell Therapy
Background:
- Endothelial progenitor cells (EPCs) exhibit reduced eNOS expression, potentially impairing their antithrombogenic capacity for vascular grafts.
- Investigating eNOS overexpression in EPCs is crucial for improving graft performance.
Purpose of the Study:
- To determine if eNOS overexpression in EPCs enhances antithrombogenic properties.
- To assess the effect of eNOS overexpression on tissue factor (TF) levels in EPCs.
Main Methods:
- Isolated CD34+ cells and differentiated them into EPCs using VEGF.
- Overexpressed eNOS via plasmid transfection in CD34+ cells.
- Analyzed TF expression, platelet adhesion in vitro, and graft thrombopoiesis/fibrin adhesion in vivo.
Main Results:
- eNOS gene transfection reduced TF levels in CD34+ cells, abrogating TNFα-induced TF expression.
- Overexpression of eNOS significantly inhibited platelet adhesion in vitro.
- In vivo studies showed decreased thrombopoiesis and fibrin adhesion on vascular grafts seeded with eNOS-overexpressing EPCs.
Conclusions:
- Overexpression of eNOS effectively decreases TF levels in EPCs.
- Enhanced eNOS expression improves the antithrombogenic properties of small-caliber vascular grafts.
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