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  本文選題：自組裝多肽 + 血管生成��； 參考：《清華大學(xué)》2012年碩士論文

【摘要】：功能化自組裝多肽納米纖維支架材料因其獨(dú)特的設(shè)計以及良好的生物相容性和可降解性已成為了新興的組織工程支架材料。三維支架材料里微血管系統(tǒng)的長入和建立是組織工程移植材料長期存活的重要保證，也是一直以來組織工程產(chǎn)品臨床應(yīng)用中所面臨的重要瓶頸之一。 本文設(shè)計和制備了一系列具有血管生成活性的自組裝多肽納米纖維水凝膠支架材料。從材料學(xué)角度分析自組裝多肽的顯微結(jié)構(gòu)，從細(xì)胞學(xué)角度篩選出能夠促進(jìn)血管生成的多肽水凝膠。為了進(jìn)一步驗(yàn)證多肽具有促血管生成作用，嘗試了內(nèi)皮細(xì)胞和平滑肌細(xì)胞的二維共培養(yǎng)，觀察細(xì)胞在多肽水凝膠上的生長情況。 從促進(jìn)內(nèi)皮細(xì)胞貼附增殖以及抑制其凋亡的蛋白質(zhì)及生長因子中，設(shè)計了六種氨基酸序列，通過多肽固相合成法將其接枝在自組裝多肽RADA16-I的末端。將1%功能化多肽與RADA16-I按1:1的體積比混合形成功能化自組裝多肽。通過圓二色譜分析其二級結(jié)構(gòu)，證實(shí)形成了β折疊結(jié)構(gòu)；在原子力顯微鏡（AFM）下可以看到均勻的納米纖維結(jié)構(gòu)；在掃描電子顯微鏡（SEM）下，可以看到其多孔狀結(jié)構(gòu)與天然的細(xì)胞外基質(zhì)結(jié)構(gòu)十分相似，是十分理想的三維組織工程支架材料。 將人臍靜脈內(nèi)皮細(xì)胞（HUVEC）種植在功能化自組裝多肽水凝膠上，通過觀察細(xì)胞的貼附、增殖、遷移及形貌分化，篩選出兩種促進(jìn)內(nèi)皮細(xì)胞增殖和形貌分化的多肽片段——KLT（KLTWQELYQLKYKGI）和PRG（GPRGDSGYRGDS），具有促血管生成的潛能。 為了進(jìn)一步證實(shí)這兩種多肽的促血管生成作用，嘗試了內(nèi)皮細(xì)胞和平滑肌細(xì)胞在二維表面上的共培養(yǎng)，，可以看到這兩種多肽具有明顯的促細(xì)胞增殖和形貌分化作用。
[Abstract]:Functional self-assembled polypeptide nanofiber scaffolds have become new scaffolds for tissue engineering because of their unique design, good biocompatibility and biodegradability.The growth and establishment of microvascular system in three-dimensional scaffolds is an important guarantee for the long-term survival of tissue engineering transplantation materials, and it is also one of the important bottlenecks in the clinical application of tissue engineering products.A series of self-assembled polypeptide nanofiber hydrogel scaffolds with angiogenic activity were designed and prepared.The microstructures of self-assembled polypeptides were analyzed from the point of view of materials, and the polypeptide hydrogels which could promote angiogenesis were screened from the perspective of cytology.In order to further verify the effect of polypeptide on angiogenesis, two dimensional co-culture of endothelial cells and smooth muscle cells was attempted to observe the growth of the cells on polypeptide hydrogel.Six amino acid sequences were designed from the proteins and growth factors that promoted the proliferation of endothelial cells and inhibited their apoptosis. They were grafted onto the end of self-assembled polypeptide RADA16-I by solid phase peptide synthesis.1% functionalized polypeptide was mixed with RADA16-I at 1:1 volume ratio to form functionalized self-assembled polypeptide.The secondary structure was analyzed by circular dichroism, and it was confirmed that the 尾 -fold structure was formed; the uniform nanofiber structure could be seen under atomic force microscope (AFM); and under scanning electron microscope (SEM),It can be seen that the porous structure is very similar to the natural extracellular matrix and is an ideal three-dimensional scaffold for tissue engineering.Human umbilical vein endothelial cells (HUVECs) were implanted on functionalized self-assembled polypeptide hydrogels. The adhesion, proliferation, migration and morphological differentiation of the cells were observed.Two kinds of polypeptide fragments, KLTWELYQLYKGI and PRGN GPRGDSGYR GDSs, were screened to promote the proliferation and morphological differentiation of endothelial cells, which have the potential of promoting angiogenesis.In order to further confirm the angiogenic effect of these two peptides, the co-culture of endothelial cells and smooth muscle cells on the two-dimensional surface was attempted. It can be seen that the two peptides have obvious effects on promoting cell proliferation and morphologic differentiation.
【學(xué)位授予單位】：清華大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R318.08

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 韓香,顧軍;多肽的固相合成[J];天津藥學(xué);2002年01期

本文編號：1751088