非病毒基因载体生物相容性研究

发布时间：2018-10-26 17:31

【摘要】：在过去二十多年间,基于基因递送的临床实验已经被广泛研究并用于治疗或者防御各种疾病。但是,载体在细胞内外的递送障碍使得临床实验的成功率很低。纳米粒子和蛋白质的相互作用是基因递送的首要障碍。药物递送系统和血液蛋白的结合被认为是载体递送至关重要的步骤,能够决定大部分的纳米粒子的生物分布、效率和毒性。本研究我们首先利用胶束/前驱体共模板组装策略制备了分子水平有机-无机杂交的硫醚桥接的介孔有机硅纳米粒子(TB-MONs),得到的纳米粒子孔径较大且粒径在30 nm左右,适宜于直接靶向细胞核。3-巯基丙基三甲氧基硅烷(MPTES)通过后接枝的方法引入到TB-MONs表面以便后续的功能化。含巯基的TB-MONs作为引发剂,在溶液中通过巯基-二硫交换的聚合在粒子表面引入细胞渗透的聚硫醚(CPD)。研究了改性前后的硫醚桥接的介孔有机硅纳米粒子的溶血效应和蛋白吸附行为。结果表明制备的TB-MONs相对于传统的MSNs蛋白吸附和溶血百分比都有所降低,且粒子表面引入CPD对粒子的蛋白吸附和溶血效应都影响不大。为了阐明结构对和蛋白相互作用的影响,我们研究了两种具有相同的分子量但是结构不同的聚阳离子及其复合体系和BSA的相互作用,这两种聚阳离子是接枝聚合物——聚(N-乙烯吡咯烷酮)-g-聚(2-甲基丙烯酸二甲氨基乙酯)(PVP-g-PDMAEMA,也就是Pg P)和嵌段聚合物——聚(N-乙烯吡咯烷酮-b-聚(2-甲基丙烯酸二甲氨基乙酯)(PVP-g-PDMAEMA,也就是Pg P)。我们利用荧光光谱来确定聚阳离子和BSA的静电结合常数和结合位点。Pg P和BSA相互作用的结合常数为8.3×10~4 M~(-1),这个值要比Pb P(4.1×10~4 M~(-1))的大,说明Pg P和BSA具有较强的相互作用。这个结论同时可以由Pb P和Pg P的结合位点值得到支持((0.3 vs 1.1)。利用表面等离子体共振传感器(SPR)来研究聚阳离子及其复合体系和BSA的非特异性相互作用,根据SPR实验的结果,两种聚阳离子和BSA的相互作用都随着浓度的增加而增加,且其复合体系和BSA的相互作用随N/P的增加而增加。
[Abstract]:Over the past two decades, gene-based clinical trials have been extensively studied and used to treat or defend against various diseases. However, the failure of delivery of carriers in and out of cells makes the success rate of clinical trials very low. The interaction of nanoparticles and proteins is the primary obstacle to gene delivery. The combination of drug delivery systems and blood proteins is considered to be a crucial step in carrier delivery, which can determine the biological distribution, efficiency and toxicity of most nanoparticles. In this study, we first prepared mesoporous organosilicon nanoparticles (TB-MONs) bridged by organic and inorganic hybrid at molecular level using micelle / precursor co-template assembly strategy. The size of mesoporous organosilicon nanoparticles (TB-MONs) was larger and the size of nanoparticles was about 30 nm. 3-mercaptopropyltrimethoxysilane (MPTES) was introduced to the surface of TB-MONs by the method of post-grafting for subsequent functionalization. TB-MONs containing sulfhydryl as initiator, polyether (CPD). Introduced into the surface of particles by mercapto-disulfide exchange polymerization in solution The hemolysis effect and protein adsorption behavior of mesoporous silicone nanoparticles bridged with sulfides were studied. The results showed that compared with the traditional MSNs protein adsorption and hemolysis percentage of the prepared TB-MONs were decreased and the effect of CPD on the protein adsorption and hemolysis of the particles was not affected by the introduction of CPD on the particle surface. In order to elucidate the effect of structure on the interaction with proteins, we studied the interaction between two polycations with the same molecular weight but different structures and their complexes with BSA. The two polycations are grafted polymers-poly (N-vinylpyrrolidone) -g- (dimethylaminoethyl 2-methacrylate) (PVP-g-PDMAEMA,) That is, Pg P) and block polymer-poly (N-vinylpyrrolidone) -b-poly (dimethylaminoethyl 2-methacrylate) (PVP-g-PDMAEMA, or Pg P). The electrostatic binding constant of polycation and BSA and the binding constant of interaction between. Pg P and BSA are 8.3 脳 10 ~ (4) M ~ (-1), which is larger than that of Pb P (4.1 脳 10 ~ (4) M ~ (-1). It shows that Pg P and BSA have strong interaction. This conclusion can be supported by the binding sites of Pb P and Pg P (0.3 vs 1.1). The surface plasmon resonance sensor (SPR) was used to study the nonspecific interaction between polycation and its composite system and BSA. According to the results of SPR experiment, the interaction between the two kinds of polycation and BSA increased with the increase of concentration. The interaction between the composite system and BSA increases with the increase of N / P.
【学位授予单位】：天津大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R450

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