本文選題：組織工程　切入點(diǎn)：多孔支架　出處：《華東理工大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：組織工程方法的基本要素包括種子細(xì)胞、支架材料和細(xì)胞因子。間充質(zhì)干細(xì)胞(Mesenchymal stem cells, MSCs)和成體細(xì)胞如軟骨細(xì)胞等都是組織工程研究中重要的種子細(xì)胞。三維多孔支架的孔徑大小以及不同化學(xué)基團(tuán)的表面修飾都可能對(duì)細(xì)胞行為有重要影響,但文獻(xiàn)報(bào)道主要集中在對(duì)材料單一性質(zhì)的研究,而將材料理化性質(zhì)相結(jié)合系統(tǒng)考察生物材料與細(xì)胞之間相互作用的研究相對(duì)匱乏。為此,本論文首先選用聚己內(nèi)酯(P olycaprolactone, PCL)材料,制備得到表面形貌光滑的PCL膜和不同孔徑(100-200μ、200-300 μm和300-450μm)的PCL三維多孔支架；然后考察不同表面化學(xué)修飾的PCL薄膜對(duì)人MSCs (human MSCs, hMSCs)貼附和生長的影響；進(jìn)一步重點(diǎn)考察了不同孔徑和表面化學(xué)修飾的PCL多孔支架材料對(duì)hMSCs的生長、遷移、增殖和分化的影響；最后,將不同密度的兔關(guān)節(jié)軟骨細(xì)胞(1abbit articular chondrocytes, rACs)接種在不同孔徑的PCL支架材料上,考察接種密度和孔徑對(duì)軟骨細(xì)胞表型維持的影響。結(jié)果發(fā)現(xiàn)：(1)通過選擇合適的化學(xué)修飾條件能夠維持PCL薄膜表面形貌,而接觸角均有降低；(2)水解處理的PCL薄膜促進(jìn)了hMSCs的貼附、鋪展和增殖,而過于劇烈的氨解處理則使PCL表面不利于hMSC s的生長；(3)化學(xué)修飾的PCL多孔支架接觸角降低,連通性良好,hMSCs可以貼著孔壁生長,也能跨孔生長,修飾后的支架更有利于細(xì)胞的貼附和增殖,而且200-300μm孔徑的支架較其它孔徑更有利于細(xì)胞貼附和增殖；(4)整體上,修飾后的支架有利于hMSCs成骨分化,PCL-H支架更能促進(jìn)hMSCs成軟骨分化,而且孔徑為200-450μm的支架更能促進(jìn)hMSCs成軟骨分化；(5)軟骨細(xì)胞在支架內(nèi)都出現(xiàn)了不同程度的去分化現(xiàn)象,接種密度為5×104 cells/scaffold比30×104 cells/scaffold的增殖速度更快,接種密度為30×104 cells/scaffold比5×104cells/scaffod和15×104 cells/scaffod對(duì)P1rACs表型的維持更有利；(6)300-450μm孔徑支架上的P1rACs的生長和糖胺聚糖(Glycosaminoglycans, GAG)的分泌較其它孔徑略大,而且細(xì)胞向支架內(nèi)部遷移較其它孔徑更深。綜上所述,多孔支架材料的表面化學(xué)性質(zhì)和孔徑尺寸都會(huì)對(duì)hMSCs的粘附、增殖和分化以及軟骨細(xì)胞表型的維持都有顯著影響,而且存在顯著的協(xié)同作用。該研究結(jié)果將可能為今后的組織構(gòu)建過程和軟骨組織工程提供科學(xué)的指導(dǎo)。
[Abstract]:The basic elements of tissue engineering methods including seed cells, scaffolds and cytokine. Mesenchymal stem cells (Mesenchymal stem cells, MSCs) and adult cells such as cartilage cells are important seed cells in the tissue engineering. Three dimensional porous scaffolds with different pore size and surface modification of chemical groups may have an important influence on the behavior of cells, but the literature reports mainly focus on research on single material properties, and the material physical and chemical properties of the system combining investigation study of the interaction between biomaterials and cells is relatively scarce. Therefore, this paper chooses polycaprolactone (P olycaprolactone, PCL) materials, preparation of PCL film surface the morphology of smooth and different aperture (100-200, 200-300 m and 300-450 m) PCL three-dimensional porous scaffold; and the effects of different surface chemical modification of PCL films on MSCs (human MSCs, HMSCs) the adhesion and growth effect; the further study on the growth of PCL porous scaffolds with different pore sizes and surface chemical modification on hMSCs migration, proliferation and differentiation; finally, the different density of rabbit articular chondrocytes (1abbit articular, chondrocytes, rACs) were seeded on PCL scaffolds with different pore sizes, influence effects of inoculum density and pore size on the phenotype of chondrocytes maintained. Results showed that: (1) by choosing appropriate chemical modification conditions to maintain the PCL film surface morphology, and the contact angle decreased; (2) PCL films attached to promote the hydrolysis of hMSCs, spreading and proliferation, and severe ammonia solution the PCL surface is not conducive to the growth of s hMSC; (3) the chemical modification of porous PCL contact angle decreased, good connectivity, hMSCs may be close to the hole wall growth, can also cross hole growth, support the modified more For cell adhesion and proliferation, but also support 200-300 m pore diameter is more conducive to cell adhesion and proliferation of other aperture; (4) on the whole, the modified scaffold for hMSCs osteogenic differentiation, PCL-H scaffolds can promote chondrogenic differentiation of hMSCs, and the aperture of 200-450 m stents can promote hMSCs chondrogenic differentiation of cartilage cells; (5) there are different degrees of dedifferentiation in stent, inoculation density of 5 * 104 cells/scaffold 30 * 104 faster than the cells/scaffold growth rate and inoculation density of 30 * 104 cells/scaffold 5 * 104cells/scaffod and 15 * 104 cells/scaffod to maintain the phenotype of P1rACs (more favorable; 6) 300-450 m aperture bracket on the growth of P1rACs and glycosaminoglycans (Glycosaminoglycans, GAG) secretion than the other slightly larger aperture, and the cell to support internal migration aperture deeper than the other. In summary, many The surface chemical properties and pore size of scaffold materials will adhere to hMSCs, have a significant impact on the proliferation and differentiation and maintain the phenotype of chondrocytes, and there is a significant synergistic effect. The results of this study may provide scientific guidance and cartilage tissue engineering for the future of the organization.

【學(xué)位授予單位】：華東理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R318.08 ;TQ317

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