CD133抗体涂层血管支架捕获造血干细胞特异性的研究

发布时间：2018-01-01 16:15

  本文关键词：CD133抗体涂层血管支架捕获造血干细胞特异性的研究　出处：《大连理工大学》2011年硕士论文　论文类型：学位论文

  更多相关文章： CD133抗体 血管支架 造血干细胞 壳聚糖 透明质酸

【摘要】：心脑血管疾病是目前危害人类生命和健康的最大杀手,血管支架术是最迅速有效的治疗手段,挽救了无数心脑血管疾病病人的生命。但第一代裸金属血管支架易产生排斥反应,导致血管再狭窄高达25%以上。第二代药物涂层支架可有效地抑制排斥反应,但也会导致远期再狭窄和其他的不良反应,如过敏反应和内膜增生等。第三代生物工程支架是通过支架携带的抗体涂层捕获外周血中修复细胞到支架表面,加速血管自然修复。 血管支架涂层载体和抗体对捕获造血干细胞(HSC)是否具有选择性是第三代生物工程血管支架的关键问题。本文建立了CD133抗体涂层血管支架抗体载量的定量分析方法,并对其进行了验证；对静电自组装制备CD133抗体涂层金属血管支架制备工艺的影响因素进行了考察；采用荧光免疫法和扫描电镜法,对壳聚糖(CH)/透明质酸(HA)基础涂层的生物相容性、CD133抗体涂层血管支架在外周血中捕获HSC的特异性以及捕获干细胞的分化结果进行评价。 结果显示：316L不锈钢冠脉血管支架和Ni-Ti合金肾动脉血管支架抗体载量分别为标示量50±15ng/支和150±20ng/支。CD133抗体涂层血管支架4℃放置半年后抗体生物活性不变。CH/HA基础涂层在血液中孵化1h后表面仍然致密,未粘附有血小板和其他有型细胞。CD133抗体涂层在外周血中仅选择性捕获鸟巢状细胞,对Hoechst33342荧光染料拒染,不表达CD20、CD7、CD61抗原,在小分子透明质酸的诱导下可特异性分化为血管内皮祖细胞(EPCs)。CD34抗体涂层在外周血中捕获的细胞有鸟巢状、纺锤状和毛球状,能被Hoechst33342荧光染料染色,并表达CD20、CD7、CD61抗原。VEGFR-2抗体涂层捕获的细胞有鸟巢状、纺锤状和有伪足卵石状,能被Hoechst33342荧光染料染色,并表达CD20、CD7、CD61抗原。CD271抗体涂层捕获的细胞有桑葚状、叠鞘状和星状,对Hoechst33342荧光染料拒染,不表达CD20、CD7、CD61抗原。 结果表明：CD133抗体涂层长期稳定性和CH/HA基础涂层生物相容性均良好。CD133抗体涂层仅捕获HSC,并且在涂层中小分子量HA的诱导下可定向分化为血管EPCs,修复损伤血管。CD34抗体涂层和VEGFR-2抗体涂层既可捕获HSC,又可捕获造血祖细胞。CD271抗体涂层仅捕获间充质干细胞,不捕获造血干/祖细胞。 综上所述,CH/HA基础涂层静电自组装负载CD133抗体涂层血管支架是一种选择性HSC捕获血管支架,优于市售的CD34抗体涂层血管支架,为更加有效地治疗心脑血管疾病提供新的手段。
[Abstract]:Cardio-cerebrovascular disease is the biggest killer of human life and health at present, vascular stenting is the most rapid and effective treatment. It has saved the lives of countless patients with cardiovascular and cerebrovascular diseases, but the first generation of bare metal vascular stents are prone to rejection. Second generation drug-coated stents can effectively inhibit rejection, but also lead to long-term restenosis and other adverse reactions. For example, allergic reaction and intimal hyperplasia. The third generation bioengineering scaffold is to capture the repair cells from peripheral blood to the surface of the stent through the antibody coating carried by the scaffold, and accelerate the natural repair of blood vessels. HSC-coated Carrier and Antibody against capture of Hematopoietic Stem cells. Selectivity is a key issue in the third generation bioengineered vascular stents. A quantitative analysis method for the antibody load of CD133 coated vascular stents was established in this paper. And it is verified; The factors influencing the preparation of metal vascular stent coated with CD133 antibody prepared by electrostatic self-assembly were investigated. The biocompatibility of chitosan / hyaluronic acid (HA) basic coating was studied by fluorescence immunoassay and scanning electron microscopy. The specificity of CD133 antibody coated stents to capture HSC in peripheral blood and the differentiation of stem cells were evaluated. The results show that:. The antibody load of 316L stainless steel coronary stent and Ni-Ti alloy renal artery stent were 50 卤15ng / branch and 150 卤20ng / branch, respectively. After placed at 4 鈩，

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