本文關(guān)鍵詞： 納米載體 診斷治療 葉酸受體 MRI造影劑 化療 靶向傳輸　出處：《中國科學技術(shù)大學》2017年博士論文　論文類型：學位論文

【摘要】：從全世界范圍看,腫瘤尤其是惡性腫瘤極大地威脅著人類的健康,如何有效的治療腫瘤、提高患者的生存質(zhì)量是腫瘤研究領(lǐng)域亟待解決的問題。腫瘤的早期診斷和治療對提高腫瘤治愈率非常關(guān)鍵。傳統(tǒng)的腫瘤診斷和治療是兩個分開的獨立過程,早期腫瘤很難被發(fā)現(xiàn),往往需要借助影像學技術(shù)輔助診斷。核磁共振成像MRI是一項先進的醫(yī)學影像技術(shù),被廣泛用于腫瘤的早期診斷,臨床50%以上的磁共振成像需要造影劑,但是臨床常用的釓基造影劑存在靶向性差、肝腎毒副作用大、體內(nèi)存留時間短等缺陷,嚴重制約了其在臨床的應用。腫瘤的治療方法包括手術(shù)治療、放射治療、化學藥物治療等,其中化學藥物治療被認為是目前臨床上最有效的治療手段。然而傳統(tǒng)的化療藥物存在組織選擇性差、藥物毒副作用大、腫瘤多耐藥性等缺點,嚴重影響了化療的效果。近年來,腫瘤"診療一體化"概念被提出,旨在將診斷和治療相結(jié)合,在提高藥物效果,減小藥物副作用的同時,監(jiān)控病變發(fā)展與治療效果,及時調(diào)整治療方案,避免對疾病的治療不足或過度治療。隨著納米技術(shù)的飛躍發(fā)展,與生物醫(yī)學相結(jié)合的納米醫(yī)藥成為研究的熱點,納米材料由于具有特殊的物理和化學性質(zhì),容易進行表面修飾和功能化設(shè)計,被用作藥物載體。本論文以腫瘤的診治為出發(fā)點,旨在構(gòu)建以納米材料為載體的腫瘤診療一體化平臺。利用納米材料尺寸和結(jié)構(gòu)的特殊性,對納米載體進行修飾改性,將抗腫瘤藥物和MR成像造影劑接載到一個納米載體中,同時完成藥物輸送和MR成像,實現(xiàn)多功能的整合。人正常組織是pH為7.4左右的中性環(huán)境,腫瘤組織為弱酸性微環(huán)境,利用這種生理差異可以設(shè)計pH敏感的納米藥物傳輸系統(tǒng),pH可控釋放藥物。腫瘤細胞表面往往特異性表達一些腫瘤標志物如葉酸受體(Folate Receptor,FR),葉酸受體是靶向治療最常用的靶點,納米給藥體系通過偶聯(lián)葉酸(Folic Acid,FA),可以特異性識別腫瘤細胞,與葉酸受體特異性結(jié)合并由受體介導內(nèi)化,完成靶向腫瘤診治。在本論文中,我們選擇了磁性氧化鐵納米材料作為藥物載體,構(gòu)建葉酸受體介導的載藥磁性納米pH敏感靶向給藥系統(tǒng)。磁性氧化鐵納米材料由于自身的超順磁性和良好的生物相容性,既可用作藥物載體又是良好的MR成像造影劑,被廣泛用于腫瘤的診斷和治療研究中。本論文內(nèi)容具體包括以下幾個部分:1.回顧腫瘤研究的文獻資料,核磁共振成像技術(shù)在腫瘤的早期診斷治療中有重要的作用,釓元素和鐵元素分別是T1、T2造影劑的理想材料選擇,已有研究證實超順磁性氧化鐵納米顆粒作為藥物載體和MR成像造影劑有很好的應用前景。在本論文的第二章中,我們對氧化鐵納米顆粒Fe304進行修飾改性,通過溶劑熱降解反應制備出釓摻雜氧化鐵納米材料GFON,由于釓離子的引入,能夠有效提高磁共振成像的T1和T2信號,是良好的T1-T2雙模式造影劑。阿霉素(Doxorubicin,DOX)是常用的廣譜抗腫瘤藥物,化療效果顯著,選擇釓鐵氧化物GFON做藥物載體,通過化學鍵在該納米載體表面接載葉酸配體FA和化療藥物阿霉DOX,構(gòu)建出兼具磁共振成像和抗腫瘤治療的多功能納米醫(yī)藥FA-GFON-DOX。材料性能檢測顯示FA-GFON-DOX納米顆粒具有良好的MR成像性能和pH敏感的藥物釋放性能;通過MTT法、流式細胞術(shù)、共聚焦實驗等細胞實驗驗證了該納米探針可以導致HeLa細胞、SCC3細胞的的凋亡或壞死,對正常的牙周膜干細胞PDLSC細胞毒性小;可以特異性識別并靶向進入HeLa細胞、SCC3細胞內(nèi)累積。2.體內(nèi)環(huán)境的復雜性,是阻礙許多納米藥物用于腫瘤治療的限制因素。為了驗證FA-GFON-DOX多功能納米給藥系統(tǒng)對在體腫瘤的成像增強和抑制效果,我們利用BALB/c免疫缺陷裸鼠分別建立了人宮頸癌HeLa細胞系、口腔鱗癌SCC3細胞系的腫瘤異種移植模型,進行了體內(nèi)實驗研究。通過荷瘤裸鼠的MR成像實驗證實FA-GFON-DOX納米探針可以顯著增強在體腫瘤的MR成像效果;荷瘤裸鼠體內(nèi)藥代動力學分析的的結(jié)果顯示,FA-GFON-DOX納米藥物可以很好的抑制宮頸癌、口腔鱗癌異種移植腫瘤的浸潤生長,減少化療藥物DOX對機體的毒副作用,提高了化療效果。3.口腔鱗癌的發(fā)病率在常見惡性腫瘤中排名第六位,由于腫瘤所在位置的特殊性很難做到早期診斷治療。化療是口腔鱗癌的主要治療手段之一,順鉑(Cisplatin,CDDP)是口腔鱗癌化療中使用最多的藥物,但是順鉑具有明顯的肝腎毒性、骨髓抑制等副作用。已有研究表明葉酸受體在鱗狀細胞癌中高度表達,因此可以選擇葉酸受體作為口腔鱗癌靶向治療的靶點。在本論文的第四章,我們致力于尋找一種新型的多功能納米探針用于口腔鱗癌的靶向診斷和治療。在第二章中,已經(jīng)通過實驗證實釓鐵氧化物納米材料是良好的藥物載體和MR成像造影劑,在本論文的第四章,通過對納米材料的進一步修飾改性制備出多孔核殼結(jié)構(gòu)的釓鐵氧化物納米顆粒CSGFN用作藥物載體,多孔核殼結(jié)構(gòu)由于比表面積高、孔隙率高可裝載更多的藥物。通過化學反應接載葉酸配體FA和抗腫瘤藥物順鉑CDDP到該納米載體上,得到葉酸受體介導的的載順鉑磁性納米pH敏感靶向給藥系統(tǒng)FA-CSGFN-CDDP用于口腔鱗癌的診斷和治療。體外實驗結(jié)果顯示,順鉑藥物CDDP的釋放是嗜酸性、pH敏感依賴型;細胞毒理實驗的結(jié)果驗證了它對口腔鱗癌SCC3細胞毒性大,流式細胞術(shù)、共聚焦實驗驗證了該納米探針可以特異性識別并進入SCC3細胞累積;我們使用BALB/c免疫缺陷裸鼠建立了口腔鱗癌SCC3細胞系的腫瘤模型,荷瘤裸鼠的MR成像實驗表明該納米探針可以顯著增強在體腫瘤的成像效果;藥物代謝實驗的結(jié)果說明FA-CSGFN-CDDP可以有效抑制SCC3細胞的異常增殖,控制口腔鱗癌的體內(nèi)浸潤發(fā)展,同時降低順鉑藥物的副作用。納米載體的使用,診療一體化概念的推出,使納米醫(yī)藥朝著多功能診斷治療的方向發(fā)展�？梢灶A測葉酸受體介導的磁性納米靶向給藥系統(tǒng)在腫瘤的MRI診斷和靶向治療方面具有巨大的應用前景,使腫瘤的可視化診治、個性化治療成為可能。
[Abstract]:From a worldwide perspective, tumor, especially malignant tumors which threaten human health, how to effectively treat tumor, improve the quality of life of patients is an urgent problem in the field of cancer research. Early diagnosis and treatment of tumors to improve the cure rate of cancer is very important. The diagnosis and treatment of tumor is the traditional two independent process separate, early cancer was found to be difficult, often need the help of imaging diagnosis. Magnetic resonance imaging MRI is an advanced medical imaging technique is widely used in the early diagnosis of cancer, clinical magnetic resonance imaging more than 50% to contrast agent, but the clinical commonly used gadolinium based contrast agent has poor targeting side effects, liver and kidney, defects such as short retention time in vivo, seriously restricts its clinical application. The treatment of cancer includes surgery, radiotherapy, chemotherapy and other treatment, The chemical medicine is considered to be the most effective treatment in clinic at present. However, the traditional chemotherapy drugs are selective tissue, drug toxicity, tumor multidrug resistance and other shortcomings, has seriously affected the effect of chemotherapy. In recent years, the tumor "theranostics" concept was proposed, aimed at diagnosis and treatment combination and in improving the drug effect, reduce the side effects of drugs at the same time, to monitor the lesion development and treatment effect, adjust treatment, avoid the treatment of disease is insufficient or excessive treatment. With the rapid development of nanotechnology, combined with biomedical nano medicine has become a hot research topic, nano materials with special physical and chemical properties easy, surface modification and functional design, was used as a drug carrier. In this paper, the diagnosis and treatment of cancer as a starting point, to construct nano materials as the carrier of the swollen The diagnosis and treatment of tumors. Using special integrated platform of nano size and structure, the modification of nano carrier, the antitumor drugs and MR imaging contrast agent to pick up a nano carrier, while the completion of drug delivery and MR imaging, to achieve the integration of multi functions. The normal group is neutral environment pH about 7.4 of the tumor tissue is weak acidic microenvironment, the physiological differences can design a pH sensitive nano drug delivery system, pH controlled release drug. Tumor cells often specific expression of some tumor markers such as folate receptor (Folate Receptor, FR), folate receptor targeted therapy is the most commonly used. Nano drug delivery system by coupling (Folic Acid, FA) of folic acid, can specifically recognize tumor cells, combined with folate receptor specificity and by receptor-mediated internalization, complete targeting tumor diagnosis and treatment. In this paper, we choose Magnetic iron oxide nano materials as drug carrier, construction of drug loaded magnetic nanoparticles of pH sensitive target folate receptor mediated drug delivery system. The magnetic iron oxide nano materials because of their superparamagnetic magnetic compatibility and good biocompatibility, which can be used as a drug carrier is good MR imaging contrast agent, is widely used in research and diagnosis the treatment of tumor. The content of this thesis includes the following parts: 1. review of tumor research literature, magnetic resonance imaging at the early stage of the tumor plays an important role in diagnosis and treatment of GD element and iron element are T1, ideal material selection T2 contrast agent, studies have confirmed superparamagnetic iron oxide nanoparticles as a drug carrier and MR imaging contrast agent has very good application prospects. In the second chapter of this thesis, we modified the iron oxide nanoparticles Fe304 by solvent thermal degradation Prepared by the reaction of GD doped iron oxide nano material GFON, due to the introduction of gadolinium ions, can effectively improve the magnetic resonance imaging of the T1 and T2 signals, is a good T1-T2 dual mode contrast agent. Adriamycin (Doxorubicin, DOX) is a commonly used broad-spectrum anti-tumor drugs, chemotherapy effect, selection of gadolinium iron oxide GFON as drug carrier and by chemical bonds in the nano surface of the carrier carrying folate ligand FA and chemotherapy drugs adriamycin DOX, constructed with magnetic resonance imaging and multifunctional nano medicine FA-GFON-DOX. materials performance testing of anti-tumor therapy showed that FA-GFON-DOX nanoparticles have good imaging performance of MR and pH sensitive drug release properties; by MTT method, flow cytometry, confocal experiment cell experimental results show that the nano probe can lead to HeLa cell, SCC3 cell apoptosis or necrosis, stem cell PDLSC cell toxicity of normal periodontal ligament; Specific recognition and targeted into HeLa cells, SCC3 cells,.2. cumulative complexity of internal environment, hinder many nano drug for tumor therapy and limitation. In order to verify the FA-GFON-DOX multifunctional nano drug delivery system for in vivo imaging enhancement and inhibition effect, we human cervical cancer HeLa cells were established by using BALB/c immunodeficient nude mice, tumor xenograft model of oral squamous cell carcinoma SCC3 cell line, was studied by MR. In vivo imaging experiment in nude mice demonstrated that FA-GFON-DOX nanoparticles can significantly enhance the MR imaging effect in tumor bearing nude mice; pharmacokinetics analysis results showed that FA-GFON-DOX nanoparticles can be very good the inhibition of cervical carcinoma, oral squamous cell carcinoma xenograft tumor invasion, reduce toxicity and side effect of DOX chemotherapy, improve the chemotherapy effect of.3. The incidence rate of oral squamous cell carcinoma in common malignant tumors ranked sixth, due to the special location of the tumor is difficult to achieve the early diagnosis and therapy. Chemotherapy is the major treatment of oral squamous cell carcinoma, cisplatin (Cisplatin, CDDP) is the most used drug in the chemotherapy of oral squamous cell carcinoma, but cisplatin has obvious liver and kidney toxicity, bone marrow inhibition of other side effects. Studies have shown that high expression of folate receptor in squamous cell carcinoma, so we can choose the folate receptor as oral squamous cell carcinoma targeted therapy targets. In the fourth chapter, we are committed to finding a new multifunctional nano probe for oral squamous cell carcinoma targeted in the diagnosis and treatment. In the second chapter, has been confirmed by gadolinium iron oxide nano materials is a good drug carrier and MR imaging contrast agent, in the fourth chapter, based on the further modification of nano materials Preparation of gadolinium iron oxide nano particles of CSGFN porous core-shell structure used as a drug carrier was modified, the porous shell structure with high specific surface area, porosity and high loading more drugs. Through chemical reaction to pick up folate ligand FA and anticancer drugs cisplatin CDDP to the nanometer carrier, get folate receptor mediated cisplatin loaded magnetic nano pH sensitive targeting drug delivery system of FA-CSGFN-CDDP for the diagnosis and treatment of oral squamous cell carcinoma in vitro. The experimental results show that cisplatin CDDP release is eosinophilic, pH sensitive type; cell toxicity test results show its high toxicity on oral squamous cell carcinoma SCC3 cells, flow cytometry, confocal experiment the nano probe can specifically recognize and enter the SCC3 cell accumulation; we use the BALB/c tumor model in immunodeficient athymic mice established oral squamous cell carcinoma SCC3 cell line in nude mice, MR like. The results show that the nanoparticles can significantly enhance the imaging effect of in vivo drug metabolism; experimental results show that FA-CSGFN-CDDP can effectively inhibit the proliferation of SCC3 cells, control of oral squamous cell carcinoma infiltration in vivo development, while reducing the side effects of cisplatin. The use of nano carrier, and launched the concept of integration, the nano medicine towards multi the function of diagnosis and treatment direction. Magnetic nanoparticles can predict target folate receptor mediated drug delivery system in MRI diagnosis and targeted therapy of tumor has great application prospect, the visualization of the tumor diagnosis, individualized treatment has become possible.

【學位授予單位】：中國科學技術(shù)大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R73-3;TB383.1

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10 趙洪偉;植物激素對口腔鱗癌誘導分化作用的實驗研究[D];四川大學;2005年

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