本文選題：表面修飾　切入點：聚己內(nèi)酯　出處：《華東理工大學(xué)》2012年碩士論文

【摘要】：材料的表面性質(zhì)不僅能影響細(xì)胞的粘附和增殖,并且會影響細(xì)胞的分化,最終可控制組織的生成。聚己內(nèi)酯(PCL)具有良好的生物相容性和生物降解性,以及優(yōu)良的機械性能和可加工性,因此,受到了廣泛關(guān)注。但是親水性差,缺乏可反應(yīng)性官能團和生物活性分子等表面性質(zhì),導(dǎo)致了材料與細(xì)胞間相互作用弱,使其在組織工程領(lǐng)域中的應(yīng)用受到了限制。因此,聚己內(nèi)酯的表面改性來提高其長期生物相容性己成為其在組織工程領(lǐng)域中應(yīng)用的熱點。 本論文旨在通過對聚己內(nèi)酯材料進(jìn)行表面改性,包括對其進(jìn)行氨基官能團修飾和引入特異性識別分子半乳糖,給細(xì)胞創(chuàng)造一個良好的人工ECM環(huán)境,模擬細(xì)胞外的基質(zhì)環(huán)境,并進(jìn)一步研究材料表面性質(zhì)的改變對細(xì)胞的調(diào)控行為。 本論文的主要工作內(nèi)容如下： 1.通過控制單體投料比合成了一系列不同含量氨基修飾的聚己內(nèi)酯材料(NPCL)。利用接觸角測試儀、X—射線光電子能譜儀和原子力顯微鏡重點考察了材料的表面親水性、表面元素組成和表面形貌。氨基的引入增加了材料的親水性并使得材料表面粗糙度增加。采用人間充質(zhì)干細(xì)胞(hMSC)為模型細(xì)胞來評估了NPCL支架材料的生物相容性,并研究了材料表面理化性能對細(xì)胞行為的影響。由于材料親水性的提高、表面拓?fù)浣Y(jié)構(gòu)的改善以及NPCL材料表面正電荷和帶負(fù)電干細(xì)胞之間存在靜電作用,NPCL材料對hMSC細(xì)胞的粘附、增殖和分化起到促進(jìn)作用,是非常具有潛力的新型組織工程支架材料。 2.以乳糖酸和NPCL聚合物為原料,在1-乙基-3(3-二甲氨丙基)碳二亞胺鹽酸鹽和N-羥基琥珀酰亞胺縮合劑的催化作用下合成了側(cè)鏈帶半乳糖的聚己內(nèi)酯(GPCL)。核磁、紅外光譜和凝膠滲透色譜法表征了聚合物的結(jié)構(gòu)和分子量并確證了乳糖酸分子偶聯(lián)成功,廣角X射線衍射和差示掃描量熱法表征了聚合物的結(jié)晶性和熱力學(xué)性能。 3.將合成的新型半乳糖修飾的GPCL材料鋪成膜,采用HepG2細(xì)胞株為模型細(xì)胞來評估半乳糖的引入對聚己內(nèi)酯材料帶來的影響。血球計數(shù)板計數(shù)法和肌動蛋白染色結(jié)合檢測接種后細(xì)胞粘附效率,CCK-8試劑盒繪制HepG2細(xì)胞增殖曲線,掃描電鏡以及Live-Dead熒光染色觀察細(xì)胞形態(tài)和死活狀況,白蛋白分泌量測試考察HepG2細(xì)胞在聚合物膜上肝功能表達(dá)狀況。GPCL上的半乳糖分子與分布在HepG2細(xì)胞表面的去唾液蛋白酶之間的特異性識別作用提高了細(xì)胞與材料間相互作用,引導(dǎo)了細(xì)胞粘附并促進(jìn)了細(xì)胞增殖及活性維持。
[Abstract]:The surface properties of the material not only affect cell adhesion and proliferation, but also affect cell differentiation, and ultimately control the formation of tissue. PCL has good biocompatibility and biodegradability. Because of its excellent mechanical properties and processability, it has attracted wide attention. However, due to its poor hydrophilicity and lack of surface properties such as reactive functional groups and bioactive molecules, the interaction between materials and cells is weak. Therefore, the surface modification of polycaprolactone to improve its long-term biocompatibility has become the focus of its application in tissue engineering. The purpose of this thesis is to create a good artificial ECM environment for cells by modifying the surface of polycaprolactone materials, including modification of amino functional groups and introduction of specific recognition molecule galactose, to simulate the extracellular matrix environment. Furthermore, the regulatory behavior of the surface properties of the materials on the cells was further studied. The main contents of this thesis are as follows:. 1. A series of polycaprolactone materials with different contents of amino modified polycaprolactone were synthesized by controlling the feed ratio of monomers. The surface hydrophilicity of the materials was investigated by means of contact angle tester, X-ray photoelectron spectroscopy (XPS) and atomic force microscope (AFM). The introduction of amino groups increased the hydrophilicity of the NPCL scaffolds and increased the surface roughness. The biocompatibility of NPCL scaffolds was evaluated by using human mesenchymal stem cells (hMSCs) as model cells. The effect of surface physical and chemical properties on cell behavior was studied. Due to the improvement of hydrophilicity, the improvement of surface topology and the existence of electrostatic interaction between positive charge and negatively charged stem cells on the surface of NPCL materials, the adhesion of NPCL materials to hMSC cells was studied. Proliferation and differentiation are potential new scaffolds for tissue engineering. 2. Polycaprolactone (GPCL) with galactose was synthesized from lactate and NPCL polymer under the catalysis of 1-ethyl-3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide condensation agent. The structure and molecular weight of the polymer were characterized by IR and gel permeation chromatography, and the molecular coupling of lactate was confirmed. The crystallization and thermodynamic properties of the polymer were characterized by wide-angle X-ray diffraction and differential scanning calorimetry. 3. The new galactose modified GPCL material was prepared. HepG2 cell line was used as model cell to evaluate the effect of galactose on the polycaprolactone material. The cell adhesion efficiency after inoculation was determined by blood cell counter counting and actin staining. The HepG2 cell proliferation curve was plotted by CCK-8 kit. Scanning electron microscopy (SEM) and Live-Dead fluorescence staining were used to observe the morphology and death of cells. The specific recognition of galactose molecules on HepG2 cells and sialytic proteases distributed on the surface of HepG2 cells enhanced the interaction between cells and materials. It leads cell adhesion and promotes cell proliferation and activity maintenance.
【學(xué)位授予單位】：華東理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R318.08

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本文編號：1700223