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研究生： 葉桓溥
論文名稱： 開發玫瑰紅裝載之複合高分子材料作為光激發藥物控制釋放之奈米平台
Development of rose bengal-loaded multipolymeric nanoplateform as photo-responsive drug carrier for cancer therapy
指導教授： 黃郁棻
口試委員: 陳仁焜
張建文
學位類別： 碩士
Master
系所名稱： 原子科學院 - 生醫工程與環境科學系
Department of Biomedical Engineering and Environmental Sciences
論文出版年： 2014
畢業學年度： 103
語文別： 中文
論文頁數： 90
中文關鍵詞： 玫瑰紅 、四氧化三鐵 、光動力治療 、藥物控制釋放
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    • 玫瑰紅(Rose Bengal)具有優良的光敏化效率，能產生大量的單態氧，於光動力治療領域深具應用潛力。過去研究可透過直接修飾如醋酸化、或藉由奈米載體攜帶，協助藥物抵達並累積於腫瘤微環境，並提升其穿越細胞膜之能力，達到有效傳遞藥物之目的。本研究主要利用甲殼素(chitosan)、聚乙烯醇(Polyvinyl alcohol)、混合短鏈聚乙烯亞胺(Polyethylenimine)，以油水乳化法將疏水性氧化鐵奈米粒子包覆於奈米團簇中，作為光動力藥物載體。此複合體除具有良好的水溶分散性，聚乙烯亞胺之陽離子聚電解質特性，亦可藉由靜電吸引力，減少負電性玫瑰紅分子於中性環境下之滲漏問題；一旦施予光照，玫瑰紅的光敏化作用會產生大量活性氧分子，進而氧化聚乙烯亞胺之三級胺集團，造成奈米團簇之表面電位下降，使玫瑰紅分子從載體中釋放，達到光動力控制藥物釋放之目的。磁性鐵奈米粒子能在本體系中進一步扮演磁共振造影對比劑的顯影功能，以及提供磁力導引靶向式藥物傳遞，利用磁性導引以及光動力誘導藥物釋放可以達到高準確性的藥物傳遞及癌症治療。
    綜合上述，本研究提出簡單而快速的合成方法，不須額外的化學性修飾以及共聚合高分子的製程，利用單純的高分子及藥物摻雜，並以靜電作用力以及親疏水作用力進行合成，完成一對氧化壓力敏感之新穎複合高分子材料，利用聚乙烯亞胺之陽離子特性，在光敏化反應中的可調控性，可控制負電性藥物或核糖核酸的吸附與釋放，開發出有別於傳統光動力治療，一智慧型之癌症治療奈米平台。

    Rose bengal (RB) which exhibits high singlet oxygen (1O2) yield is a clinical promising photosensitizer in anticancer therapy. Several RB hydrophobic derivatives (e.g., acetate) have been developed to facilitate intracellular accumulation. Nanoparticles also serve as effective carriers to deliver these compounds to the tumor microenvironment, or to cross biological barriers such as cell membranes. In this study, RB molecules were encapsulated in a mixture of chitosan, polyvinyl alcohol and branched polyethylenimine (bPEI) with hydrophobic iron oxide nanoparticles through an oil-in-water emulsion method. The as-prepared multipolymeric magnetic nanoclusters displayed high water dispersibility and the cationic groups of bPEI were effective for RB loading through electrostatic interaction. In addition, triggered release of the loaded drugs also occurred simultaneously during the photodynamic reaction. The improvement of photodynamic-stimulated triggered release holds great promise in precise control of drug delivery.

    摘要 II Abstract III 致謝 IV 第一章　緒論 1 1.1奈米科技與生醫應用 1 1.1.1奈米材料簡介 1 1.1.2奈米藥物載體 2 1.1.3觸發-應答式藥物載體 4 1.2癌症以及藥物治療 10 1.2.1癌症簡介 10 1.2.2癌症的藥物治療 11 1.2.3光動力治療 12 1.3超順磁奈米氧化鐵鐵於癌症治療應用 17 1.3.1超順磁奈米氧化鐵 17 1.3.2超順磁奈米氧化鐵的表面修飾材料 18 1.3.3超順磁奈米氧化鐵的粒子構型與物理特性 21 1.3.4超順磁奈米氧化鐵於藥物載體開發 22 1.4研究動機與目的 24 第二章　實驗材料與方法 27 2.1 實驗藥品與儀器 27 2.1.1 實驗藥品 27 2.1.2 緩衝溶液配置 29 2.1.3 細胞培養與操作 30 2.1.4 儀器 30 2.2 四氧化三鐵奈米粒子的合成與特性鑑定 32 2.2.1四氧化三鐵奈米粒子合成 32 2.2.2四氧化三鐵奈米粒子的鑑定 32 2.3藥物載體之合成與鑑定 33 2.3.1高分子四氧化三鐵奈米團簇合成 33 2.3.2玫瑰紅裝載高分子四氧化三鐵奈米團簇合成 34 2.3.3 藥物載體之載附效率定量 35 2.3.4 藥物載體之特性鑑定 36 2.4藥物載體溶液條件的特性探討 36 2.4.1溶液條件的自由基產率分析 36 2.4.2溶液條件的藥物洩漏分析 37 2.4.3溶液條件的藥物釋放分析 37 2.4.4藥物載體於不同環境之穩定性探討 37 2.4.5光照對藥物載體的特性影響 37 2.4.6活性氧自由基對藥物載體的特性影響 38 2.4.7活性氧自由基對高分子氧化的影響 38 2.4.8 RB對bPEI的光氧化行為分析 38 2.5藥物載體與目標細胞 MCF-7 的作用 39 2.5.1流式細胞儀分析載體吞噬效率 39 2.5.2流式細胞儀分析載體自由基產生之效率 39 2.5.3普魯士藍染色分析載體於細胞吞噬效率 40 2.5.4共軛交螢光顯微鏡分析細胞內藥物釋放 41 2.5.5細胞存活率分析 41 2.5.6外加磁場對載體以及細胞作用的影響 42 第三章　實驗結果與討論 44 3.1 四氧化三鐵奈米粒子的合成與特性鑑定 44 3.1.1四氧化三鐵奈米粒子合成與鑑定 44 3.2藥物載體之合成與鑑定 44 3.2.1高分子四氧化三鐵奈米團簇合成與鑑定 44 3.2.2藥物載體於不同環境之穩定性探討 46 3.2.3 藥物載體之載附效率定量 46 3.3藥物載體溶液條件的特性探討 47 3.3.1溶液條件的自由基產率分析 47 3.3.2溶液條件的藥物洩漏分析 48 3.3.3溶液條件的藥物釋放分析 48 3.3.4藥物載體之氧化壓力應答性討論 49 3.4藥物載體與目標細胞 MCF-7 的作用 51 3.4.1流式細胞儀分析載體吞噬效率及自由基產生之效率 51 3.4.2探討藥物載體於細胞內的自由基生成效率 52 3.4.3顯微鏡分析載體於細胞吞噬效率 52 3.4.4共軛交螢光顯微鏡分析細胞內藥物釋放 53 3.4.5細胞存活率分析 53 3.5奈米藥物載體的磁物理特性探討 55 3.5.1飽和磁化量分析 55 3.5.2核磁共振T2造影對比能力分析 55 3.5.3高分子藥物載體的熱重分析 55 第四章　結論 57 第五章 未來展望 59 圖表說明 60 參考文獻 84

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