本文選題：磁性納米顆粒　切入點(diǎn)：胰腺癌　出處：《蘇州大學(xué)》2014年碩士論文

【摘要】：胰腺癌早期癥狀不顯著，早期診斷缺乏特異性，因此死亡率高。近年來，分子影像學(xué)發(fā)展迅速，其以分子生物學(xué)為基礎(chǔ)，在活體狀態(tài)下從分子、基因水平對(duì)胰腺癌進(jìn)行更早期的診斷。磁共振成像（magnetic resonance imaging，，MRI）具有空間分辨率高、組織對(duì)比度好等優(yōu)點(diǎn)，因此在分子影像中應(yīng)用廣泛，但敏感性較低，因此限制了其在腫瘤早期診斷的作用。近紅外熒光成像（Near-infrared fluorescence imaging, NIRF）具有靈敏度高、無(wú)射線輻射、對(duì)人體無(wú)害、可重復(fù)曝光等優(yōu)點(diǎn)，但是空間分辨率較差，背景噪聲高。同時(shí)利用MRI和熒光成像對(duì)同一物體進(jìn)行成像以獲得補(bǔ)充信息，即MRI/NIRF雙模態(tài)成像，可以實(shí)現(xiàn)優(yōu)勢(shì)互補(bǔ)，其研究的重點(diǎn)是雙模態(tài)成像探針的制備。故研制新型的磁性熒光納米顆粒，并探討其性能特征及其在胰腺癌雙模態(tài)成像中的應(yīng)用，是本文的主要工作。 本論文主要內(nèi)容分為以下兩個(gè)部分：（1）制備雙模態(tài)探針的前體-磁性納米顆粒（2）制備雙模態(tài)探針并探討其生物安全性、體內(nèi)生物分布及其在胰腺癌的靶向成像效果。 第一部分主要是制備磁性納米顆粒。首先是油性的超順磁性氧化鐵納米顆粒（superparamagnetic iron oxide nanoparticles, SPIONs）的制備，實(shí)驗(yàn)以Fe(acac)3為前驅(qū)體，以油酸、油胺、1,2-十六烷以及二芐醚的混合物為表面活性劑，在有機(jī)溶液中，用高溫?zé)峤夥ㄖ苽溆腿苄缘腟PIONs。并用TEM、XRD、VSM等方法對(duì)其進(jìn)行表征，結(jié)果表明所制備的油溶性SPIONs具有良好的形貌、均一的尺寸，并具有超順磁性，無(wú)剩磁。然后用兩種不同的方法對(duì)所合成的油溶性SPIONs進(jìn)行表面改性。第一種方法基于雙親性改性，先用反相旋蒸法使親水性大分子PMAO取代油溶性SPIONs表面的疏水性配體，然后通過2,2'-(乙烯二氧)雙(乙胺)和PMAO的開環(huán)聚合反應(yīng)，得到足夠的氨基、羧基用于后期的熒光染料及抗體的偶聯(lián)。第二種方法是配體交換，先利用NOBF4將油溶性SPIONs轉(zhuǎn)換為易溶于DMF等的親水性顆粒，然后用BSA通過配體交換將親水性納米顆粒轉(zhuǎn)換為水溶性的BSA·SPIONs。TEM、膠體穩(wěn)定性及弛豫率測(cè)試表明這兩種方法修飾合成的納米顆粒均具有良好的水溶性、膠體穩(wěn)定性及較高的弛豫率。 第二部分通過MTT實(shí)驗(yàn)和組織HE染色考察BSA·SPIONs的生物安全性，結(jié)果表明該納米粒子對(duì)人肝正常上皮L02細(xì)胞的毒性較低，具有良好的生物相容性。納米粒子在體內(nèi)的生物分布，是通過小鼠尾靜脈注射BSA·SPIONs后，在不同時(shí)間點(diǎn)取出相應(yīng)的組織器官，測(cè)定其內(nèi)Fe的含量來進(jìn)行的。研究結(jié)果顯示該納米顆粒進(jìn)入體內(nèi)后主要分布于肝、脾、大腸，于注射后6-12h達(dá)到高峰，其后逐漸下降至正常水平。BSA·SPIONs偶聯(lián)近紅外熒光染料Cy5及靶向胰腺癌的抗Plec-1抗體后，即靶向的雙模態(tài)探針dyeBSA·SPION-mAb，用其進(jìn)行細(xì)胞水平的MRI、近紅外熒光成像及在體MRI，驗(yàn)證了該探針較低的非特異性吸附、良好的靶向性及用于MRI/NIRF雙模態(tài)成像的可行性。 綜上，本文用兩種方法制成了具有良好的生物相容性及穩(wěn)定性的磁性納米顆粒，選用BSA修飾的磁性顆粒偶聯(lián)熒光染料后，初步探究了該探針的生物安全性及體內(nèi)生物分布。將該探針偶聯(lián)靶向抗體后，可用作腫瘤早期探測(cè)，并通過體外、體內(nèi)靶向雙模態(tài)成像的研究，證明了該新型磁性熒光雙模態(tài)探針的生物醫(yī)學(xué)應(yīng)用價(jià)值，說明其廣闊的應(yīng)用前景。
[Abstract]:The early symptoms of pancreatic cancer is not significant, early diagnosis and lack of specificity, therefore the mortality rate is high. In recent years, the rapid development of molecular imaging, which is based on molecular biology, in vivo from molecular gene level for diagnosis of pancreatic cancer earlier. Magnetic resonance imaging (magnetic resonance, imaging, MRI) with spatial resolution the advantages of high tissue contrast and so on, so the application of molecular imaging in wide range, but the sensitivity is low, thus limiting its role in early diagnosis of cancer. Near infrared fluorescence imaging (Near-infrared fluorescence, imaging, NIRF) has the advantages of high sensitivity, no radiation, harmless to the human body, can be repeated exposure and other advantages, but the space low resolution, high background noise. At the same time using MRI and fluorescence imaging imaging of the same object in order to obtain additional information, namely MRI/NIRF dual mode imaging, can achieve complementary advantages, its research The emphasis is focused on the preparation of dual mode imaging probes. Therefore, the development of new magnetic fluorescent nanoparticles and their performance characteristics and their application in dual mode imaging of pancreatic cancer are the main works of this paper.
The main contents of this paper are divided into two parts: (1) preparation of dual modal probe precursor magnetic nanoparticles (2) to prepare bimodal probes and explore their biological safety, in vivo biodistribution and targeted imaging effect in pancreatic cancer.
The first part is the preparation of magnetic nano particles. The first is oily superparamagnetic iron oxide nanoparticles (superparamagnetic iron oxide nanoparticles, SPIONs) were prepared based on Fe (ACAC) 3 as precursor, oleic acid, oil amine, 1,2- sixteen alkyl benzyl ether mixture and two as surfactant in the organic solution, using high temperature pyrolysis preparation of oil soluble SPIONs. and TEM, XRD, VSM and other methods were characterized. Results show that the preparation of oil soluble SPIONs has good appearance, uniform size, and superparamagnetism, no residual surface modification of the synthetic oil soluble SPIONs and then using two different methods. The first method is based on amphiphilic modified the hydrophilic macromolecular PMAO to replace the oil soluble hydrophobic ligands of SPIONs surface by rotary evaporation method, and then 2,2'- (two ethylene oxide) - (ethylamine) open loop and PMAO The polymerization reaction, get enough amino, carboxyl for coupling fluorescent dye and later antibody. The second method is ligand exchange, using NOBF4 oil soluble SPIONs converted into hydrophilic particles easily dissolve in DMF, and ligand exchange hydrophilic nanoparticles convert soluble BSA - SPIONs.TEM by water with BSA, colloid stability and relaxation rate test showed that the two kinds of methods to modify the synthesis of nanoparticles have good water solubility, colloid stability and higher relaxation rate.
The second part was the biological safety of SPIONs BSA was investigated by MTT experiment and HE, results showed that the toxicity of nanoparticles on human normal liver epithelial L02 cells is low, has good biocompatibility. Nanoparticles in vivo biodistribution, through tail vein injection of BSA SPIONs, remove the corresponding organization organ at different time points, the determination of Fe in the paper. The results of research showed that the nano particles into the body mainly distributed in liver, spleen, intestine, after injection of 6-12h reached the peak, then decreased gradually to the normal level of.BSA and SPIONs coupling near infrared fluorescent dye Cy5 and anti Plec-1 targeting pancreatic cancer the antibody, targeting the dual mode probe dyeBSA - SPION-mAb cell level with its MRI, near-infrared fluorescence imaging and in vivo MRI, verified the nonspecific adsorption of the probe is low, targeted and good The feasibility of MRI/NIRF dual mode imaging.
In summary, this paper use two methods made of magnetic nanoparticles has good biological compatibility and stability of magnetic particles modified by BSA coupling fluorescent dye, preliminary study of the probe biological safety and biodistribution. The probe coupling targeting antibodies, can be used for early tumor detection, and through in vitro, to study the dual mode imaging in vivo, proved that the new biomedical applications of magnetic fluorescent double mode probe value shows its broad application prospects.

【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R735.9;R445.2

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