本文選題：肝臟 + 去細(xì)胞��； 參考：《瀘州醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的：文章通過肝素修飾的方法，對(duì)改善作為組織工程支架材料的去細(xì)胞化豬肝臟的生物相容性進(jìn)行了初步研究，，以期望能將此方法用于改善組織工程肝臟材料的生物相容性，以及為其他血液接觸的組織工程材料改善生物相容性提供一個(gè)參考辦法。方法：我們用不同的肝素修飾方法制作了不同種類的肝素化去細(xì)胞化豬肝臟支架材料。t-LBL （tissue immobilized with heparin by ionic binding vialayer-by-layer self-assembly technique）支架材料是通過去細(xì)胞化豬肝臟支架經(jīng)肝素層層自組裝離子結(jié)合的方式完成的。t-MPA(tissueimmobilized with heparin by covalent binding via multi-point attachment)支架材料是通過去細(xì)胞化豬肝臟支架經(jīng)肝素多點(diǎn)連接共價(jià)結(jié)合的方式完成的。t-EPA(tissue immobilized with heparin by covalent bindingvia end-point attachment)支架材料是通過去細(xì)胞化豬肝臟支架經(jīng)末端連接共價(jià)結(jié)合的方式完成的。文章通過甲苯胺藍(lán)染色（Toluidine bluestaining）檢測(cè)、甲苯胺藍(lán)比色法（toluidine-blue colorimetric method）檢測(cè)、H E染色（Hematoxylin Eosin staining）檢測(cè)、凝血時(shí)間檢測(cè)（coagulation time tests）、血小板粘附實(shí)驗(yàn)（Platelet adhesion）、血漿復(fù)鈣時(shí)間檢測(cè)（Plasma recalcification time）、材料上HepG2細(xì)胞培養(yǎng)實(shí)驗(yàn)（Cell culture of HepG2）、MTT分析、溶血度檢測(cè)實(shí)驗(yàn)（Haemolysisratio measurement）和掃描電鏡（scanning electron microscopy，SEM）檢測(cè)評(píng)估了肝素化去細(xì)胞化豬肝臟支架材料的肝素化效果、抗凝血性能、細(xì)胞相容性。結(jié)果：t-LBL, t-MPA和t-EPA肝素化去細(xì)胞豬肝臟支架材料擁有不同的肝素含量。肝素化的去細(xì)胞化豬肝臟支架材料比未肝素化的去細(xì)胞豬肝臟支架材料具有更好的抗凝血性能。和t-LBL, t-MPA肝素化去細(xì)胞支架材料比較，t-EPA肝素化去細(xì)胞支架材料擁有更多肝素、肝素釋放速度更慢而保留的肝素比例更多。和t-LBL, t-MPA肝素化去細(xì)胞豬肝臟支架材料比較，t-EPA肝素化去細(xì)胞豬肝臟支架材料的凝血酶時(shí)間（the thrombin time，TT），凝血酶原時(shí)間（prothrombin time，PT），活化部分凝血活酶時(shí)間（activatedpartial thromboplastin time，APTT）超出了設(shè)備測(cè)量范圍。而且，t-EPA肝素化去細(xì)胞豬肝臟支架材料的血漿復(fù)鈣時(shí)間是肝素修飾各組支架材料中最長(zhǎng)的。t-EPA肝素化去細(xì)胞豬肝臟支架材料比對(duì)照、t-LBL和t-MPA更能促進(jìn)HepG2細(xì)胞在材料上增殖、分布、聚集和伸展。結(jié)論：通過末端（end-point attachment，EPA）連接肝素共價(jià)結(jié)合修飾的去細(xì)胞化豬肝臟支架材料較層層自組裝（layer-by-layerself-assembly technique，LBL）和多點(diǎn)結(jié)合（multi-point attachment，MPA）肝素化去細(xì)胞化豬肝臟支架材料獲得了良好的抗凝性和細(xì)胞相容性。鑒于在溫和的水溶液條件下完成肝素修飾的去細(xì)胞化豬肝臟支架材料制備和其在肝臟組織工程中擁有良好的抗凝性能，本研究為制備其他組織工程用的具有良好抗凝性能的與血液接觸的支架材料提供了一種參考方法。
[Abstract]:Objective: to improve the biocompatibility of decellated porcine liver as tissue engineering scaffold material by heparin modification method in order to improve the biocompatibility of tissue engineering liver materials. And to provide a reference for other blood contact tissue engineering materials to improve biocompatibility. Methods: different kinds of heparinized porcine liver scaffolds. T-LBL tissue immobilized with heparin by ionic binding vialayer-by-layer self-assembly techniques were prepared by different heparin modification methods. The. T-MPAtissue with heparin by covalent binding via multi-point attachment material, completed by the method of assembling ion binding, is made by means of multipoint connection of heparin and covalent binding of porcine liver stents. T-EPA-tissue immobilized with heparin by covalent bindingvia end-point attachment materials are fabricated by means of the. T-EPA-tissue immobilized with heparin by covalent bindingvia end-point attachment material, which is made by means of decellulization of porcine liver stents by means of multipoint connection and covalent binding of heparin. The acellular porcine liver scaffold was covalently linked to the end of the liver. Toluidine bluestainingwas detected by toluidine blue staining, and Hematoxylin eosin stainingwas detected by toluidine blue colorimetry and toluidine blue colorimetric method. Coagulation time test, platelet adhesion test, plasma recalcification assay, cell culture of HepG2 cell culture assay were used. The hemolysis ratio measurement and scanning electron microscopy (SEM) were used to evaluate the heparin effect, anticoagulant performance and cytocompatibility of heparinized porcine liver scaffolds. Results different heparin contents were found in 10% t-LBL, t-MPA and t-EPA heparinized porcine liver scaffolds. Heparinized porcine liver scaffold has better anticoagulant performance than non-heparinized porcine liver scaffold. Compared with t-LBLand t-MPA heparinized scaffolds, the heparinized stents had more heparin, slower heparin release rate and more heparin retention. Compared with t-LBLand t-MPA heparinized porcine liver scaffolds, the thrombin time and prothrombin time of t-EPA heparinized porcine liver scaffold materials were higher than those of the thrombin thromboplastin time, and the activated partial thromboplastin time (APTT) exceeded the measuring range of the equipment. Moreover, the plasma recalcification time of heparinized porcine stents was the longest. T-EPA heparinized porcine liver scaffolds could promote the proliferation and distribution of HepG2 cells. Gather and stretch. Conclusion: the anticoagulant and cytocompatibility of heparin covalently modified porcine liver scaffolds by end-point attachment (EPA) is better than that of layer-by-layerself-assembly technique (LBLL) and multi-point attachment MPA-heparinized porcine liver scaffolds. In view of the preparation and anticoagulant properties of heparin modified acellular porcine liver scaffolds in mild aqueous solution, This study provides a reference method for the preparation of other scaffolds with good anticoagulant properties for tissue engineering.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R318.08;R657.3

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 文鵬飛;宋克東;劉天慶;朱艷霞;;殼聚糖支架材料的制備、表面修飾及細(xì)胞粘附性能的研究[J];高�；瘜W(xué)工程學(xué)報(bào);2008年06期

2 郭利明;曾小飛;馬瑞東;尚觀勝;郝明;易定華;;去細(xì)胞豬主動(dòng)脈瓣RGD肽表面修飾促進(jìn)細(xì)胞黏附的實(shí)驗(yàn)研究[J];四川大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2010年06期

3 趙微加;魏麗;何江虹;楊宇民;;京尼平固定NGF殼聚糖膜的制備及緩釋NGF作用探討[J];交通醫(yī)學(xué);2010年03期

4 ;The hemocompatibility and the reabsorption function of TiO_2 nanotubes biomembranes[J];Chinese Science Bulletin;2012年16期

5 顧漢卿;組織工程研究中的若干問題[J];中華整形外科雜志;2001年01期

6 陳景華,林樅,顧其勝,徐政;制備透明質(zhì)酸衍生物交聯(lián)劑的應(yīng)用研究[J];中國(guó)修復(fù)重建外科雜志;2004年01期

7 周敏;劉長(zhǎng)建;衛(wèi)志慶;劉昭;蔣雪峰;喬彤;冉峰;;bFGF預(yù)載脫細(xì)胞支架構(gòu)建小口徑人工血管的研究[J];中國(guó)修復(fù)重建外科雜志;2008年03期

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