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研究生: 張翊菁
Chang, Yi-Ching
論文名稱: 以芬頓反應驅動高分子囊泡進行超音波顯影與治療
A Fenton Reaction Activable Polymersome in Ultrasound-mediated Imaging and Therapy
指導教授: 葉晨聖
Yeh, Chen-Sheng
學位類別: 碩士
Master
系所名稱: 理學院 - 化學系
Department of Chemistry
論文出版年: 2016
畢業學年度: 104
語文別: 中文
論文頁數: 97
中文關鍵詞: 高分子囊泡超音波活性氧化物質 (ROS)
外文關鍵詞: Polymersomes, Ultrasound, Reactive oxygen species (ROS)
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  • 超音波是一種非侵入性的診斷與治療工具,具有方便使用、低成本、與即時性觀測等優點,近年來許多科學家積極發展多功能超音波顯影劑,提升超音波於生物醫學的應用價值。本研究提出一種同時具有超音波顯影與治療功能的材料,此材料是以雙重乳化的方式合成,外殼為聚乳酸-羥基乙酸共聚物 (poly(lactic-co-glycolic acid), PLGA) 鑲嵌四氧化三鐵奈米粒子,中心包覆雙氧水 (H2O2),作為氣體的來源,增強超音波影像的對比。材料在超音波環境下會破裂,釋放包覆於中心的雙氧水,雙氧水與鑲嵌於聚乳酸-羥基乙酸共聚物殼層中的四氧化三鐵奈米粒子接觸,發生芬頓反應 (Fenton reaction) 產生活性氧化物質 (Reactive oxygen species, ROS) ,利用活性氧化物質對細胞的毒殺效果達到癌症治療的目的。因為鑲嵌在聚乳酸-羥基乙酸共聚物殼層中的四氧化三鐵奈米粒子具有磁響應之特性與磁振造影的功能,因此可藉由磁導引的方式將材料導引至腫瘤部位產生磁振影像,再給予超音波刺激誘導材料破裂進行活性氧化物質介導的癌症治療,由實驗結果證實本材料於生物影像之觀測與惡性腫瘤的治療都有很好的應用價值。

    We demonstrate a successful synthesis of a theranostic polymersomes and provide a series of imaging and therapy platform in this article. We synthesis the theranostic polymersomes by double emulsion (water/oil/water) method. The core area of the polymersomes is hydrophilic, and the space between the core and outer shell is hydrophobic. The hydrophobic ligand iron oxide nanoparticles can be embedded in the hydrophobic space. Due to the component of Fe3O4 nanoparticles which provides ferromagnetic property, it can be applied as T2-negative contrast agents of magnetic resonance imaging (MRI). We can use magnet targeting the tumor site to guide the polymersomes to tumor. Combining with ultrasound irradiate, this material can get ultrasound imaging and induced ROS mediated therapy. The in vivo test provide a strong evidence for cancer treatment.

    摘要 I 英文延伸摘要 (Extended Abstract) II 誌謝 X 目錄 XII 表目錄 XVII 圖目錄 XVIII 第一章 緒論 1 1-1 超音波 (Ultrasound) 1 1-1-1 超音波的簡介 1 1-1-2 超音波的產生 2 1-1-3 音波的運動 4 1-1-4 超音波成像原理 5 1-1-5 超音波治療原理 7 1-1-6 超音波顯影劑 8 1-1-7 超音波結合顯影劑於疾病治療上之應用 11 1-2 磁振造影 (Magnetic Resonance Imaging, MRI) 16 1-2-1 磁振造影的原理 16 1-2-2 磁振造影顯影劑 18 1-3 高分子囊泡 (polymersomes) 20 1-3-1 高分子囊泡的介紹 20 1-3-2 高分子囊泡的合成方法 21 1-4芬頓反應 (Fenton reaction) 24 1-4-1 芬頓反應的簡介 24 1-4-2 芬頓反應於生物醫學之應用 25 1-5 活性氧化物質 (Reactive oxygen species, ROS) 27 1-5-1 活性氧化物質的簡介 27 1-5-2 活性氧化物質於生物醫學上的應用 29 第二章 實驗藥品與儀器設備 31 2-1 實驗藥品 31 2-1-1 合成高分子囊泡之化學藥品 31 2-1-2 細胞實驗所須之化學藥品 32 2-1-3 實驗細胞株 33 2-1-4 實驗動物 33 2-2 儀器設備 34 第三章 以芬頓反應驅動高分子囊泡進行超音波顯影與治療 37 3-1 研究動機與目的 37 3-2 材料設計與合成 39 3-2-1 油相四氧化三鐵 (Fe3O4) 奈米粒子之製備 40 3-2-2 水相四氧化三鐵奈米粒子 (Fe3O4@PLGA) 之製備 41 3-2-3 Fe3O4-PLGA高分子囊泡之合成 42 3-2-4 H2O2/PLGA高分子囊泡之合成 43 3-2-5 H2O2/Fe3O4-PLGA高分子囊泡之合成 44 3-2-6 高分子囊泡之表面修飾 46 3-3 材料分析 47 3-3-1 以碘化鉀 (KI) 觀察雙氧水的包覆 47 3-3-2 以蛋白質分析套組定量修飾在高分子囊泡表面的牛血清白蛋白 48 3-3-3 以Aminophenyl fluorescein (APF) 偵測氫氧自由基 50 3-3-4 以無機化合物Ru(dpp)3Cl2偵測氧氣 52 3-3-5 超音波實驗 53 3-3-6 高分子囊泡穩定性測試 53 3-3-7 超音波破壞高分子囊泡之機制探討 53 3-3-8 高分子囊泡之毒性分析 54 3-4 材料應用 57 3-4-1 以H2O2/Fe3O4-PLGA高分子囊泡作為顯影劑之動物超音波影像 57 3-4-2 以H2O2/Fe3O4-PLGA高分子囊泡作為顯影劑之動物磁振造影 59 3-4-3 以超音波驅動H2O2/Fe3O4-PLGA高分子囊泡產生活性氧自由基治療惡性腫瘤 60 3-4-4高分子囊泡在生物體內的分布、組織切片與血液分析 61 第四章 實驗結果與討論 62 4-1 材料鑑定 62 4-1-1 四氧化三鐵 (Fe3O4) 奈米粒子 62 4-1-2 高分子囊泡 (polymersomes) 64 4-1-3 高分子囊泡之中空結構 67 4-1-4 高分子囊泡之表面修飾牛血清白蛋白 68 4-1-5 以碘化鉀 (KI) 觀察雙氧水的包覆 69 4-1-6以Aminophenyl fluorescein (APF) 偵測氫氧自由基 71 4-1-7以無機化合物Ru(dpp)3Cl2偵測氧氣 75 4-1-8 體外超音波影像實驗 76 4-1-9高分子囊泡穩定性測試 77 4-1-10生物相容性測試 79 4-2 機制探討 80 4-2-1超音波破壞高分子囊泡之機制 80 4-3 H2O2/Fe3O4-PLGA高分子囊泡作為顯影劑與腫瘤治療 81 4-3-1 超音波驅動之細胞毒殺實驗 81 4-3-2 體內超音波影像實驗 82 4-3-3 體內磁振造影 85 4-3-4以超音波驅動產生活性氧自由基治療惡性腫瘤 87 4-3-5 高分子囊泡在生物體內的分布、組織切片與血液分析 89 第五章 結論 92 參考文獻 93

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