本文關(guān)鍵詞： 聚乙烯醇 卡拉膠 生物活性 定向支架 組織工程　出處：《天津大學(xué)》2016年博士論文　論文類型：學(xué)位論文

【摘要】：組織工程支架的設(shè)計(jì)應(yīng)該按照仿生原理,最大限度地模擬目標(biāo)組織或器官的細(xì)胞外基質(zhì)的組成、結(jié)構(gòu)和功能,以達(dá)到修復(fù)創(chuàng)傷和重建功能的目的。天然關(guān)節(jié)軟骨組織具有獨(dú)特的結(jié)構(gòu),這與其功能性密切相關(guān),本文結(jié)合組織仿生學(xué)理論,優(yōu)化材料組成和性能,并對(duì)支架進(jìn)行仿生設(shè)計(jì)和研究。具體研究?jī)?nèi)容如下:1.聚乙烯醇(PVA)水凝膠具有與天然關(guān)節(jié)軟骨類似多孔結(jié)構(gòu)和力學(xué)性能,適合于關(guān)節(jié)軟骨修復(fù)的研究。但純PVA水凝膠不僅缺乏細(xì)胞粘附性,而且成型過(guò)程中容易出現(xiàn)形貌皺縮的現(xiàn)象,嚴(yán)重限制了其在組織工程領(lǐng)域中的應(yīng)用。本文通過(guò)加入天然多糖卡拉膠(CAR)組分,利用冰凍-融化循環(huán)技術(shù)得到了PVA與CAR的復(fù)合水凝膠(CP),結(jié)果表明所制得的CP水凝膠在冷凍干燥過(guò)程中具有抗皺縮性能,表現(xiàn)出良好的結(jié)構(gòu)穩(wěn)定性。同時(shí),由于CAR組分對(duì)PVA結(jié)晶性產(chǎn)生影響,得到的復(fù)合材料內(nèi)部結(jié)構(gòu)相對(duì)疏松,明顯提高了CP凝膠材料的孔徑;2.對(duì)CP復(fù)合材料進(jìn)行生物學(xué)評(píng)價(jià),體外細(xì)胞實(shí)驗(yàn)結(jié)果表明CAR的加入可以顯著提高PVA水凝膠的生物活性,促進(jìn)細(xì)胞在材料上的粘附和增殖,并且能夠維持細(xì)胞的良好表型;通過(guò)體外溶血實(shí)驗(yàn)以及巨噬細(xì)胞激活實(shí)驗(yàn)表明,CP復(fù)合材料不會(huì)引起溶血,保持很低的巨噬細(xì)胞激活率,具有潛在的體內(nèi)應(yīng)用前景。3.利用定向冰凍-融化循環(huán)技術(shù),得到具有取向結(jié)構(gòu)的PVA-CAR定向支架,仿生了天然軟骨的組織結(jié)構(gòu),具有優(yōu)異的理化性能。體外細(xì)胞實(shí)驗(yàn)表明,定向支架相對(duì)非定向支架,更有利于細(xì)胞的遷移和增殖,有效地提高了細(xì)胞在支架內(nèi)部的均勻分布,保證支架結(jié)構(gòu)的均勻穩(wěn)定。同時(shí),體內(nèi)組織相容性實(shí)驗(yàn)結(jié)果表明支架的取向結(jié)構(gòu)賦予其良好的滲透性,能夠引導(dǎo)宿主細(xì)胞的長(zhǎng)入和增殖,為新生組織提供適宜的生存環(huán)境,促進(jìn)組織與支架之間的相互作用;4.以PVA和CAR為基體,添加無(wú)機(jī)組分羥基磷灰石(HA),制備了HA-CP復(fù)合支架材料,在優(yōu)選支架材料理化性能基礎(chǔ)上對(duì)其進(jìn)行生物學(xué)性能評(píng)價(jià),結(jié)果表明HA-CP支架表現(xiàn)出良好的細(xì)胞與組織相容性;在此基礎(chǔ)上,采用滲透與定向冰凍-融化循環(huán)相結(jié)合的技術(shù),得到OCP/HA-CP雙層復(fù)合支架材料,其上層具有取向結(jié)構(gòu)用于構(gòu)架軟骨層,下層為多孔結(jié)構(gòu)用于構(gòu)建骨層,模擬了關(guān)節(jié)軟骨與軟骨下骨的特點(diǎn)。提高滲透時(shí)間可以提高雙層支架的界面結(jié)合強(qiáng)度,有效地防止上下層的分離,有希望解決以往因存在明顯的界面而導(dǎo)致軟骨與軟骨下骨難以結(jié)合的問(wèn)題。
[Abstract]:Tissue engineering scaffolds should be designed to maximize the composition, structure and function of the extracellular matrix of the target tissue or organ in accordance with the bionic principle. Natural articular cartilage has a unique structure, which is closely related to its function. In this paper, the material composition and properties are optimized based on tissue bionics theory. And the biomimetic design and study of the scaffold. The specific research contents are as follows: 1. Polyvinyl alcohol PVA) hydrogel has similar porous structure and mechanical properties to natural articular cartilage. It is suitable for the repair of articular cartilage, but the pure PVA hydrogel is not only lack of cell adhesion, but also prone to the appearance of shrinkage during the molding process. The application of Carrageenan in tissue engineering was severely restricted. In this paper, the natural polysaccharide carrageenan carrageenan (Carrageenan) component was added. The composite hydrogels of PVA and CAR were obtained by freezing and melting cycle technique. The results show that the prepared hydrogels have anti-shrinkage property during freeze-drying process. At the same time, due to the effect of CAR composition on the crystallinity of PVA, the internal structure of the composite is relatively loose, which obviously improves the pore size of CP gel material. 2. The biological evaluation of CP composite material in vitro showed that the addition of CAR could significantly improve the biological activity of PVA hydrogel and promote cell adhesion and proliferation on the material. And can maintain the good phenotype of cells; In vitro hemolysis test and macrophage activation test showed that CP composite did not cause hemolysis and maintained a very low rate of macrophage activation. The PVA-CAR scaffold with oriented structure was obtained by using the technique of directional freeze-thawing cycle, and the tissue structure of natural cartilage was simulated. 3. In vitro cell experiments show that the directional scaffold is more conducive to cell migration and proliferation than the non-directional scaffold and can effectively improve the uniform distribution of cells in the scaffold. At the same time, the results of in vivo histocompatibility experiment showed that the oriented structure of the scaffold has good permeability and can guide the growth and proliferation of host cells. To provide suitable living environment for newborn tissue and promote the interaction between tissue and scaffold. 4. HA-CP composite scaffolds were prepared by adding hydroxyapatite as inorganic component and PVA and CAR as matrix. The biological properties of HA-CP scaffolds were evaluated on the basis of physicochemical properties. The results showed that HA-CP scaffolds showed good cell and histocompatibility. On this basis, the OCP/HA-CP bilayer composite scaffold material was obtained by using the technique of osmosis and directional freeze-melting cycle. The upper layer of the composite scaffold has a oriented structure for the framework cartilage layer. The lower layer is a porous structure to construct the bone layer, which simulates the characteristics of articular cartilage and subchondral bone. Increasing the osmotic time can improve the interface bonding strength of the double-layer scaffold and effectively prevent the separation of the upper and lower layers. It is hopeful to solve the problem that cartilage and subchondral bone are difficult to combine because of obvious interface.
【學(xué)位授予單位】：天津大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R318.08

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