超声波辅助点击反应制备形貌可控的可双点击聚合物纳米粒子及其生物学应用的研究

发布时间：2018-07-16 15:56

【摘要】：聚合物纳米粒子(PNPs)在生物学领域有着广泛的应用研究价值,得益于其优良的生物相容性和易于功能化修饰的优点,多功能PNPs已被广泛应用于药物输送、细胞显影、生物分析和聚合物治疗等方面。而PNPs的形貌对其行为和性质有巨大的影响,因此其形貌控制一直是聚合物纳米材料领域的关键点之一。本研究通过超声波辅助点击聚合的方法不仅实现了对PNPs形貌的控制(即呈现纳米球或者纳米棒的形状),同时利用该方法制备的PNPs表面残留的点击基团为其在温和条件下进一步特别功能化提供了极大的方便;并据此制备了生物相容性良好的靶向糖纳米粒子和荧光糖纳米粒子,分别应用于糖-蛋白相互作用和细胞显影方面的研究。1、本研究利用超声波辅助无皂点击聚合方法合成表面具有双重可点击基团的纳米粒子(DCNPs),并通过系统研究单体结构、溶剂、催化剂种类、超声波频率以及功率对PNPs形貌的影响,实现对DCNPs形貌的控制。结果表明在超声波辐射下(40~100kHz),以Cu(PPh)3Br为催化剂、乙腈/氯仿(96:4,v/v)混合物为溶剂可以得到粒度分布较窄的纳米球;而在40 kHz条件下,以氯仿为溶剂得到形状规整的聚合物纳米棒。纳米棒形成机理的研究表明,催化剂Cu(PPh)3Br中铜离子和聚合物分子链中吡啶环上氮原子之间的络合作用,是聚合物形成规整的棒状结构的重要因素。对纳米球表面残留的炔基和叠氮基进行定量表征发现,1 g聚合物中含有38μmol的叠氮基和15μmol的炔基,因此特别便于其在温和条件下的进一步的功能化修饰。2、为了证明本方法合成的DCNPs的可修饰性及展示其潜在的生物应用,我们以纳米球为例,利用点击反应分别制备了糖纳米粒子(GNPs)和荧光糖纳米粒子(FGNPs):1)通过将叠氮基和炔基修饰的乳糖分子分别修饰到纳米粒子表面,形成生物相容性好的糖纳米粒子,然后利用石英晶体微天平技术研究糖-蛋白相互作用。结果显示无论是与修饰在芯片表面的半乳糖凝集素作用,还是直接与固定在芯片表面的Jurkat细胞作用,GNPs都显示出很高的特异结合活性。2)通过将炔基化的乳糖分子和叠氮基修饰的罗丹明分子修饰到纳米球表面形成荧光糖纳米粒子(FGNPs),并用于细胞成像的研究。激光共聚焦图像显示该荧光糖纳米粒子在与Jurkat细胞共培养30 min后就可以被细胞摄取到细胞内部。这种荧光糖纳米粒子探针的构建模式很适合用于构建凝集素靶向给药系统。可以预见,本研究制备的DCNPs在糖生物学、细胞成像和生物医药方面具有广泛的潜在应用价值。
[Abstract]:Polymer nanoparticles (PNPs) have been widely used in the field of biology. Due to their excellent biocompatibility and easy functional modification, multifunctional PNPs have been widely used in drug delivery and cell development. Biological analysis and polymer therapy. The morphology of PNPs has a great influence on its behavior and properties, so its morphology control has always been one of the key points in the field of polymer nanomaterials. In this study, ultrasonic assisted click polymerization not only controlled the morphology of PNPs (that is, the shape of nanospheres or nanorods), but also made use of the clickable groups on the surface of PNPs prepared by this method. Further special functionalization under the conditions provides great convenience; Based on this, targeted sugar nanoparticles and fluorescent sugar nanoparticles with good biocompatibility were prepared. In this study, ultrasonic assisted soap-free click polymerization method was used to synthesize double clickable nanoparticles (DCNPs) with double clickable groups, and the monomer structure and solvent were systematically studied. The influence of the type of catalyst, ultrasonic frequency and power on the morphology of PNPs can be realized by controlling the morphology of DCNPs. The results show that nanospheres with narrow particle size distribution can be obtained by using Cu (PPH) 3BR as catalyst and acetonitrile / chloroform (96: 4 v / v) as solvent under ultrasonic irradiation (40kHz), while polymer nanorods with regular shape can be obtained by using chloroform as solvent at 40kHz. The formation mechanism of nanorods shows that the complexation between copper ions in Cu (PPH) 3BR catalyst and nitrogen atoms on pyridine ring in polymer chains is an important factor in the formation of regular rod structure of polymers. The residual acetylene and azide groups on the surface of the nanospheres were quantitatively characterized. It was found that there were 38 渭 mol azide groups and 15 渭 mol acetylene groups in the 1 g polymer. In order to prove the modifiability of DCNPs synthesized by this method and to demonstrate its potential biological applications, we take nanospheres as an example. Sugar nanoparticles (GNPs) and fluorescent sugar nanoparticles (FGNPs) (FGNPs: 1) were prepared by click-through reaction to form biocompatible sugar nanoparticles by modifying lactose molecules modified by azide group and alkynyl group onto the surface of nanoparticles, respectively. Then the sugar-protein interaction was studied by quartz crystal microbalance. The results showed that the effect of galactose agglutinin modified on the surface of the chip, Or directly interacting with Jurkat cells immobilized on the surface of the chip, GNPs showed high specific binding activity (.2) by modifying acetylated lactose molecules and azide modified Rhodamine molecules onto the surface of the nanospheres to form fluorescein. Rice particles (FGNPs), and used in the study of cell imaging. Laser confocal images showed that the fluorescent sugar nanoparticles could be absorbed into Jurkat cells after co-culture for 30 min. The fluorescent sugar nanoparticles probe is suitable for the construction of lectin targeting drug delivery system. It can be predicted that the DCNPs prepared in this study have a wide range of potential applications in sugar biology, cell imaging and biomedicine.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TB383.1;O631.5

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