本文選題：血液相容性　切入點(diǎn)：內(nèi)皮化　出處：《西南交通大學(xué)》2012年博士論文

【摘要】：內(nèi)皮祖細(xì)胞(Endothelial progenitor cell, EPC)對(duì)于血管支架植入后損傷血管內(nèi)皮細(xì)胞層的重建具有重要作用。目前,新開發(fā)的內(nèi)皮祖細(xì)胞捕獲血管支架相對(duì)于藥物洗脫支架和裸支架具有更好的治療效果,但仍存在一些問題：1)由于未考慮抗凝血修飾,血栓仍可能導(dǎo)致治療失敗；2)血液中內(nèi)皮祖細(xì)胞數(shù)量稀少,且在血流剪切力作用下,難以實(shí)現(xiàn)在支架植入部位的快速粘附,阻礙了內(nèi)皮細(xì)胞層快速重建；3)目前血管支架材質(zhì)為不可降解金屬,其長(zhǎng)期存在于血管中擾亂了血管舒縮等正常生理功能,為晚期血栓和晚期支架內(nèi)再狹窄埋下隱患。 EPC對(duì)心肌梗死患者的壞死心肌細(xì)胞再生和心肌功能恢復(fù)具有重要作用,但由于其在病變部位粘附困難,難以實(shí)現(xiàn)對(duì)病變部位的有效治療,所以將EPC定向運(yùn)輸?shù)街付ú课坏姆椒▽W(xué)研究具有重要的學(xué)術(shù)意義和潛在的應(yīng)用價(jià)值。 針對(duì)以上問題,本文采用：1)在硅烷修飾的純鈦表面接枝雙羧基聚乙二醇后,再進(jìn)一步共價(jià)固定CD34抗體,構(gòu)建出具備優(yōu)異血液相容性和特異性捕獲EPC的多功能生物活性表面；2)制備了具有可以特異性結(jié)合EPC的MNP,用于將結(jié)合MNP的EPC,在外加磁場(chǎng)輔助下,富集于磁靶向部位(支架植入處或是心肌梗死區(qū)域)并加速EPC粘附,實(shí)現(xiàn)病變部位快速EPC覆蓋；3)研究了可生物降解的鐵支架在MNP和外磁場(chǎng)輔助下表面快速粘附EPC的可能性。具體內(nèi)容如下： 1.純鈦依次經(jīng)堿熱處理和硅烷修飾,在表面獲得大量的高反應(yīng)活性的氨基,并通過氨基-羧基縮合將雙羧基的聚乙二醇接枝在純鈦表面,通過反應(yīng)條件的控制,獲得不同鏈長(zhǎng)和不同鏈密度PEG接枝表面,通過多種檢測(cè)手段證明表面構(gòu)建成功；詳細(xì)研究了PEG鏈長(zhǎng)和鏈密度與表面生物相容性的關(guān)系,結(jié)果顯示具有高鏈密度(3chains/nm2)和長(zhǎng)鏈(PEG4000)的PEG接枝表面可以更有效抑制纖維蛋白原、血小板、平滑肌細(xì)胞和巨噬細(xì)胞的粘附。 2.在對(duì)PEG接枝表面優(yōu)選后,通過PEG上的羧基與CD34抗體上的氨基縮合,實(shí)現(xiàn)CD34抗體在表面的固定,多種檢測(cè)手段證明表面構(gòu)建成功,對(duì)構(gòu)建表面的生物相容性的研究結(jié)果顯示,在PEG長(zhǎng)鏈接枝的鈦表面連接抗體后,不但保持了優(yōu)異的血液相容性和捕獲EPC的能力,而且表現(xiàn)出對(duì)平滑肌細(xì)胞粘附的抑制作用和更低的炎癥反應(yīng)。 3.通過水熱共沉淀法合成出表面包裹雙羧基聚乙二醇的Fe304,并進(jìn)一步共價(jià)連接CD34抗體,制備出具有良好磁響應(yīng)性和對(duì)EPC特異性識(shí)別的磁性納米粒子(MNP),結(jié)果顯示該MNP以100μg/mL的使用濃度為宜,該濃度下EPC在磁場(chǎng)施加部位粘附數(shù)量成倍提升而平滑肌細(xì)胞粘附被抑制,且未引發(fā)凝血和溶血,動(dòng)物體內(nèi)實(shí)驗(yàn)證明其在體內(nèi)亦具有對(duì)EPC的特異性結(jié)合能力和對(duì)磁場(chǎng)的響應(yīng)能力,并不會(huì)被免疫系統(tǒng)快速清除。 4.可降解鐵支架在體內(nèi)出現(xiàn)了嚴(yán)重的再狹窄,但其降解特性嚴(yán)重阻礙了表面生物化修飾,故難以解決再狹窄問題,通過MNP和外加磁場(chǎng)輔助,保證其降解行為不受影響的前提下有效實(shí)現(xiàn)了EPC在表面的快速粘附,結(jié)果顯示高磁場(chǎng)強(qiáng)度,長(zhǎng)磁場(chǎng)作用時(shí)間,高EPC濃度可以增加EPC在鐵表面粘附量。對(duì)于鐵支架的應(yīng)用具有重要參考價(jià)值。
[Abstract]:Endothelial progenitor cells ( EPC ) play an important role in the reconstruction of vascular endothelial cells after vascular stent implantation . Currently , newly developed endothelial progenitor cells capture vascular stents with better therapeutic effects relative to drug eluting stents and bare stents , but there are still some problems : 1 ) Thrombus may still lead to treatment failure due to the absence of anticoagulant modification ;
2 ) the number of endothelial progenitor cells in the blood is sparse , and under the action of blood flow shearing force , it is difficult to realize the rapid adhesion of endothelial progenitor cells at the implantation site of the stent , thereby inhibiting the rapid reconstruction of the endothelial cell layer ;
3 ) At present , the vascular stent material is non - degradable metal , and the long - term presence of the vascular stent material is disturbed by the normal physiological function of the vasomotor and the like in the blood vessel , and the hidden trouble is hidden in the late thrombosis and the late stent restenosis .

EPC plays an important role in the regeneration of necrotic myocardial cells and the recovery of myocardial function in patients with myocardial infarction , but it is difficult to achieve effective treatment of the lesion site due to the difficulty of adhering to the lesion site , so the methodological research for transporting the EPC to the designated site has important academic significance and potential application value .

In view of the above problems , 1 ) After grafting the double carboxyl polyethylene glycol on the silane - modified pure titanium surface , we further covalently immobilize the CD34 antibody to construct a multifunctional bioactive surface with excellent blood compatibility and specific capture EPC .
2 ) preparing MNP which can specifically bind to the EPC , and is used for enriching the EPC of the MNP , and enriching the EPC with the magnetic targeting site ( the stent implantation or the myocardial infarction area ) under the assistance of the external magnetic field and accelerating the EPC adhesion to realize the rapid EPC coverage of the lesion site ;
3 ) The possibility of rapid adhesion of the biodegradable iron stent to the surface of MNP and external magnetic field is studied . The specific contents are as follows :

1 . the pure titanium is sequentially subjected to alkali heat treatment and silane modification , a large amount of high - reactive amino groups are obtained on the surface , and the polyethylene glycol of the bicarboxyl group is grafted on the surface of the pure titanium by amino - carboxyl condensation , the PEG grafting surfaces with different chain lengths and different chain density are obtained through the control of the reaction conditions , and the surface construction success is proved by various detection means ;
The relationship between PEG chain length and chain density and surface biocompatibility was studied in detail . The results showed that PEG grafted surface with high chain density ( 3chains / nm2 ) and long chain ( PEG4000 ) could inhibit the adhesion of fibrinogen , platelets , smooth muscle cells and macrophages more effectively .

2 . After the PEG grafting surface is preferred , the immobilization of the CD34 antibody on the surface is realized by the amino condensation on the carboxyl group on the PEG and the amino group on the CD34 antibody , and various detection means prove that the surface construction is successful , and the research result of the biocompatibility of the construction surface shows that the excellent blood compatibility and the ability of capturing the EPC are maintained after the antibody is attached to the titanium surface of the PEG long linking branch , and the inhibition effect on the adhesion of the smooth muscle cell and the lower inflammatory reaction are shown .

3 . The magnetic nanoparticles ( MNP ) with good magnetic responsiveness and EPC - specific recognition were synthesized by hydrothermal coprecipitation method . The results showed that the concentration of EPC was better than that of 100 渭g / mL . The results showed that the adhesion of EPC to 100 渭g / mL was higher than that of EPC under the concentration of 100 渭g / mL .

4 . The degradable iron stent has serious restenosis in the body , but its degradation characteristics seriously hinder the surface biological modification . Therefore , it is difficult to solve the problem of restenosis . By means of MNP and external magnetic field , the rapid adhesion of the EPC to the surface is effectively realized . The results show that the high magnetic field strength , the long magnetic field action time and the high EPC concentration can increase the adhesion amount of the EPC to the iron surface .

【學(xué)位授予單位】：西南交通大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R318.0

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