非病毒基因載體生物相容性研究
發(fā)布時(shí)間：2018-10-26 17:31
【摘要】：在過去二十多年間,基于基因遞送的臨床實(shí)驗(yàn)已經(jīng)被廣泛研究并用于治療或者防御各種疾病。但是,載體在細(xì)胞內(nèi)外的遞送障礙使得臨床實(shí)驗(yàn)的成功率很低。納米粒子和蛋白質(zhì)的相互作用是基因遞送的首要障礙。藥物遞送系統(tǒng)和血液蛋白的結(jié)合被認(rèn)為是載體遞送至關(guān)重要的步驟,能夠決定大部分的納米粒子的生物分布、效率和毒性。本研究我們首先利用膠束/前驅(qū)體共模板組裝策略制備了分子水平有機(jī)-無機(jī)雜交的硫醚橋接的介孔有機(jī)硅納米粒子(TB-MONs),得到的納米粒子孔徑較大且粒徑在30 nm左右,適宜于直接靶向細(xì)胞核。3-巰基丙基三甲氧基硅烷(MPTES)通過后接枝的方法引入到TB-MONs表面以便后續(xù)的功能化。含巰基的TB-MONs作為引發(fā)劑,在溶液中通過巰基-二硫交換的聚合在粒子表面引入細(xì)胞滲透的聚硫醚(CPD)。研究了改性前后的硫醚橋接的介孔有機(jī)硅納米粒子的溶血效應(yīng)和蛋白吸附行為。結(jié)果表明制備的TB-MONs相對(duì)于傳統(tǒng)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對(duì)粒子的蛋白吸附和溶血效應(yīng)都影響不大。為了闡明結(jié)構(gòu)對(duì)和蛋白相互作用的影響,我們研究了兩種具有相同的分子量但是結(jié)構(gòu)不同的聚陽離子及其復(fù)合體系和BSA的相互作用,這兩種聚陽離子是接枝聚合物——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)。我們利用熒光光譜來確定聚陽離子和BSA的靜電結(jié)合常數(shù)和結(jié)合位點(diǎn)。Pg P和BSA相互作用的結(jié)合常數(shù)為8.3×10~4 M~(-1),這個(gè)值要比Pb P(4.1×10~4 M~(-1))的大,說明Pg P和BSA具有較強(qiáng)的相互作用。這個(gè)結(jié)論同時(shí)可以由Pb P和Pg P的結(jié)合位點(diǎn)值得到支持((0.3 vs 1.1)。利用表面等離子體共振傳感器(SPR)來研究聚陽離子及其復(fù)合體系和BSA的非特異性相互作用,根據(jù)SPR實(shí)驗(yàn)的結(jié)果,兩種聚陽離子和BSA的相互作用都隨著濃度的增加而增加,且其復(fù)合體系和BSA的相互作用隨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.
【學(xué)位授予單位】：天津大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R450
本文編號(hào)：2296463
[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.
【學(xué)位授予單位】：天津大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R450
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 Ning Zhang;Chong-Xi Wang;Jin-Hao Liu;Jin-Feng Xing;An-Jie Dong;;A kind of modified bovine serum albumin with great potential for applying in gene delivery[J];Chinese Chemical Letters;2013年07期
，本文編號(hào)：2296463
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