本文關(guān)鍵詞：CD133抗體涂層血管支架捕獲造血干細(xì)胞特異性的研究　出處：《大連理工大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： CD133抗體 血管支架 造血干細(xì)胞 殼聚糖 透明質(zhì)酸

【摘要】：心腦血管疾病是目前危害人類生命和健康的最大殺手,血管支架術(shù)是最迅速有效的治療手段,挽救了無數(shù)心腦血管疾病病人的生命。但第一代裸金屬血管支架易產(chǎn)生排斥反應(yīng),導(dǎo)致血管再狹窄高達(dá)25%以上。第二代藥物涂層支架可有效地抑制排斥反應(yīng),但也會導(dǎo)致遠(yuǎn)期再狹窄和其他的不良反應(yīng),如過敏反應(yīng)和內(nèi)膜增生等。第三代生物工程支架是通過支架攜帶的抗體涂層捕獲外周血中修復(fù)細(xì)胞到支架表面,加速血管自然修復(fù)。 血管支架涂層載體和抗體對捕獲造血干細(xì)胞(HSC)是否具有選擇性是第三代生物工程血管支架的關(guān)鍵問題。本文建立了CD133抗體涂層血管支架抗體載量的定量分析方法,并對其進(jìn)行了驗證；對靜電自組裝制備CD133抗體涂層金屬血管支架制備工藝的影響因素進(jìn)行了考察；采用熒光免疫法和掃描電鏡法,對殼聚糖(CH)/透明質(zhì)酸(HA)基礎(chǔ)涂層的生物相容性、CD133抗體涂層血管支架在外周血中捕獲HSC的特異性以及捕獲干細(xì)胞的分化結(jié)果進(jìn)行評價。 結(jié)果顯示：316L不銹鋼冠脈血管支架和Ni-Ti合金腎動脈血管支架抗體載量分別為標(biāo)示量50±15ng/支和150±20ng/支。CD133抗體涂層血管支架4℃放置半年后抗體生物活性不變。CH/HA基礎(chǔ)涂層在血液中孵化1h后表面仍然致密,未粘附有血小板和其他有型細(xì)胞。CD133抗體涂層在外周血中僅選擇性捕獲鳥巢狀細(xì)胞,對Hoechst33342熒光染料拒染,不表達(dá)CD20、CD7、CD61抗原,在小分子透明質(zhì)酸的誘導(dǎo)下可特異性分化為血管內(nèi)皮祖細(xì)胞(EPCs)。CD34抗體涂層在外周血中捕獲的細(xì)胞有鳥巢狀、紡錘狀和毛球狀,能被Hoechst33342熒光染料染色,并表達(dá)CD20、CD7、CD61抗原。VEGFR-2抗體涂層捕獲的細(xì)胞有鳥巢狀、紡錘狀和有偽足卵石狀,能被Hoechst33342熒光染料染色,并表達(dá)CD20、CD7、CD61抗原。CD271抗體涂層捕獲的細(xì)胞有桑葚狀、疊鞘狀和星狀,對Hoechst33342熒光染料拒染,不表達(dá)CD20、CD7、CD61抗原。 結(jié)果表明：CD133抗體涂層長期穩(wěn)定性和CH/HA基礎(chǔ)涂層生物相容性均良好。CD133抗體涂層僅捕獲HSC,并且在涂層中小分子量HA的誘導(dǎo)下可定向分化為血管EPCs,修復(fù)損傷血管。CD34抗體涂層和VEGFR-2抗體涂層既可捕獲HSC,又可捕獲造血祖細(xì)胞。CD271抗體涂層僅捕獲間充質(zhì)干細(xì)胞,不捕獲造血干/祖細(xì)胞。 綜上所述,CH/HA基礎(chǔ)涂層靜電自組裝負(fù)載CD133抗體涂層血管支架是一種選擇性HSC捕獲血管支架,優(yōu)于市售的CD34抗體涂層血管支架,為更加有效地治療心腦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 鹵15ng / branch and 150 鹵20ng / branch, respectively. After placed at 4 鈩，

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