Department of Biomedical Engineering, Duke University, Durham, North Carolina 27708-0281, USA.
Tissue Eng Part A. 2011 Aug;17(15-16):2091-100. doi: 10.1089/ten.TEA.2010.0631. Epub 2011 May 25.
This study investigated the augmentation of endothelial progenitor cell (EPC) thromboresistance by using gene therapy to overexpress thrombomodulin (TM), an endothelial cell membrane glycoprotein that has potent anti-coagulant properties. Late outgrowth EPCs were isolated from peripheral blood of patients with documented coronary artery disease and transfected with an adenoviral vector containing human TM. EPC transfection conditions for maximizing TM expression, transfection efficiency, and cell viability were employed. TM-overexpressing EPCs had a fivefold increase in the rate of activated protein C production over native EPCs and EPCs transfected with an adenoviral control vector expressing β-galactosidase (p<0.05). TM upregulation caused a significant threefold reduction in platelet adhesion compared to native EPCs, and a 12-fold reduction compared to collagen I-coated wells. Additionally, the clotting time of TM-transfected EPCs incubated with whole blood was significantly extended by 19% over native cells (p<0.05). These data indicate that TM-overexpression has the potential to improve the antithrombotic performance of patient-derived EPCs for endothelialization applications.
本研究通过基因治疗过表达血栓调节蛋白（TM）来增强内皮祖细胞（EPC）的抗血栓形成能力，TM 是一种内皮细胞膜糖蛋白，具有很强的抗凝血特性。从已确诊冠心病患者的外周血中分离出晚期生长 EPC，并转染含有人 TM 的腺病毒载体。采用了最大限度提高 TM 表达、转染效率和细胞活力的 EPC 转染条件。与天然 EPC 和转染表达β-半乳糖苷酶的腺病毒对照载体的 EPC 相比，TM 过表达的 EPC 激活蛋白 C 的产生速度增加了五倍（p<0.05）。与天然 EPC 相比，TM 的上调使血小板黏附减少了三分之二，与胶原 I 涂层孔相比减少了 12 倍。此外，与天然细胞相比，与全血孵育的 TM 转染的 EPC 的凝血时间延长了 19%（p<0.05）。这些数据表明，TM 过表达有可能改善患者来源的 EPC 的抗血栓形成性能，用于内皮化应用。
