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研究生:許名孜
研究生(外文):Hsu, Ming-Tzu
論文名稱:可穿透之脂質體修飾硫化銅/金銀三角奈米板載體透過對流增強遞送系統應用於腦瘤治療
論文名稱(外文):Penetrated Delivery of Lipid-Coated Triangular CuS-tagged Ag@Au Core/Shell Nanoplates to Brain Tumor through Convection-Enhanced Delivery
指導教授:胡尚秀
指導教授(外文):Hu, Shang-Hsiu
口試委員:姜文軒陳冠宇彭志剛
口試委員(外文):Chiang, Wen-HsuanChen, Guan-Yu
口試日期:2020-08-28
學位類別:碩士
校院名稱:國立清華大學
系所名稱:生醫工程與環境科學系
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:2020
畢業學年度:109
語文別:英文
論文頁數:84
中文關鍵詞:金銀合金奈米粒子硫化銅脂質體免疫佐劑磁電效應對流增強遞送系統腦瘤治療
外文關鍵詞:Ag@Au nanoplatesCuS nanocrystalsliposomeImmunoadjuvantmagnetoelectric effectsconvection-enhanced deliverybrain cancer therapy
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雖然醫療逐漸進步,但比起以往人們罹患癌症的機率也提高。近年來,癌症也躍升為國人十大死因之首,如何積極面對癌症治療並研發出高效率的治療方式已增加病人癒後的生活品質已經是不容忽視的問題。
其中因為腦區獨特的組織—血腦障壁的存在,雖然保護外來物質不易侵入腦區維持其正常的運作,卻也為藥物遞送帶來治療上的困難。現今臨床上的腦癌治療方式,包括:手術切除、放射治療與化學藥物治療…等方式,術後病人常有嘔吐、疲倦無力、腹瀉…等副作用,受到血腦障壁的阻隔,許多方式受到限制,比起其他癌症,治療效果也有限。憑藉著世界各地的專家們的努力,藥物遞送系統如火如荼得發展,當今炙手可熱的小分子藥物自然是各國關注鑽研的對象。許多研究學者們紛紛開發奈米藥物載體,透過標靶藥物治療結合藥物控制釋放的特性,提高藥物在腫瘤區的累積已達到治療效果。即便如此,仍有絕大部分得分子被血腦障壁擋住,預期成效因此大打折扣。因此唯有同時增加小分子藥物療效及找到穿透血腦障壁的方法,雙管齊下,才能達到治療效果,也藉此提高病人癒後的生活機能,希望降低其身體負擔。
因此,在本研究中,我們合成出具有「可穿透之脂質體修飾硫化銅/金銀三角奈米板載體」平台,包覆脂質體以提高藥物水溶性及生物相容性,並裝載含有含有免疫調節功能的免疫佐劑 Resiquimod (R-848) 的次級載體-樹狀聚合物。載體的部分首先利用Ag作為犧牲模板,接著加入聚乙烯吡咯烷酮(PVP ) 作為用於調節晶體生長尺寸的封端劑,並以四氯合金酸(HAuCl4)作為Au源,經由TEM的分析,確定成功合成出大小約90-120nm且具穩定性的Ag@Au奈米板。接著以熱裂解法同樣以TEM鑑定,確定合成出大小約20 nm的硫化銅,將兩者結合不但增加奈米粒子的接觸面積同時也降低硫化銅的生物毒性並解決其尺寸太小容易被生物降解的問題。
另外,選擇對流增強遞送系統 ( Convection-enhanced delivery, CED)作為輸送藥物的方式,如此一來可以直接穿越血腦障壁並準確注射藥物載體進入腫瘤區,同時提高載體在腫瘤區的累積量及藥物濃度,提高治療效果。在動物實驗上,透過CED的方式,使得奈米粒子在腫瘤區有良好的分布,且因為流速控制得當,並未產生逆流的現象。對小鼠施以高週波治療後,藉由磁電效應誘發免疫反應,在腫瘤區T細胞有明顯增加的情形,達到預期結果。
總結「可穿透之脂質體修飾硫化銅/金銀三角奈米板載體」為一個具有前瞻性的治療平台,除了本身具有熱療效外,裝載R-848免疫佐劑可以增加其免疫反應,有效達到腫瘤抑制的目標,為往後腦瘤治療開闢新的機會。


關鍵字:金銀合金奈米粒子、硫化銅、脂質體、免疫佐劑、磁電效應
、對流增強遞送系統、腦瘤治療
With the progress of nanotechnology, there are more and more treatment approaches in the recent years. However, the brain tumor is still regarded as a troublesome disease to treat because of its characterization of high mortality and high recurrence. In this study, we successfully combine Ag@Au nanoplates with copper sulfide (CuS) nanocrystals as carriers, and modify with liposome to increase the solubility of them. Moreover, we consider dendrimers as secondary drug carrier which can deliver the immune modulator (R848) in order to enhance the immune response, and increase penetration of nanoparticles into the target sites. Injecting nanoparticles through convection-enhance delivery (CED) system can both across directly to the blood-brain barrier (BBB) and increase the accumulation of carriers in the brain tumor. The outcome suggests that the nanoparticles we design present low toxicity and high cellular uptake efficiency, and can be triggered by the high-frequency magnetic field (HFMF). By doing so, we finally stop brain cancer cells from growing fast and extend the survival days of mice. It is a promising and potential nanoparticle in the drug delivery system.

Key words:Ag@Au nanoplates, CuS nanocrystals, liposome, Immunoadjuvant, magnetoelectric effects, convection-enhanced delivery, brain cancer therapy
中文摘要 I
Abstract III
Table of contents IV
List of Scheme VII
List of Figure VIII
Chapter 1 Introduction 1
Chapter 2 Literature review and theory 3
2.1 Brain cancer 3
2.1.1 The blood-brain barrier (BBB) 4
2.2 Convection- enhanced delivery system 5
2.2.1 Introduction of Convection-enhanced delivery (CED) system 6
2.2.2 Convection-enhanced delivery system as the treatment of brain tumors 8
2.2.3 Cather design 10
2.3 Nanoparticles as drug delivery system 13
2.3.1 The particle size effect 16
2.4 Nanocomposites targeted tumor delivery system 18
2.5 Characteristics of Gold-silver alloy nanoparticles 19
2.5.1 Gold-silver alloy nanoparticles reduce the toxicity of silver 24
2.5.2 Application of gold-silver alloy nanoparticle in photothermal therapy 25
2.6 Characteristics and application of copper sulfide (CuS) nanoparticles 28
2.7 Cancer Immunotherapy 30
2.7.1 Immunotherapy of brain cancer 31
2.8 The magnetic nanoparticles property and cancer applications 34
2.9 Dendrimer 35
Chapter 3 Experimental section 37
3.1 Materials 37
3.2 Apparatus 39
3.3 Method 41
3.3.1 Synthesis of Ag nanoplates (seeds) 41
3.3.2 Preparation of Growth Solution of Au 41
3.3.3 Synthesis of Triangular Ag@Au Core/Shell Nanoplates 42
3.3.4 Synthesis of Copper Sulfide (CuS) nanoctystals 43
3.3.5 Synthesis of Ag@Au-CuS 44
3.3.6 Synthesis of Ag@Au-CuS@lipo.-Den-R848 nanoparticles 44
3.3.7 Characterization 46
3.3.8 Analysis of Ag@Au core/shell nanoplates 47
3.3.9 Cell culture 47
3.3.10 Cellular Uptake 47
3.3.11 Cell viability assay 49
3.3.12 Penetration of the nanoparticles in ALTS1C1 spheroids 49
3.3.13 In vivo experiments 51
Chapter 4 Results and Discussions 53
4.1 Synthesis and characterization of Ag nanoplates, Ag@Au triangular nanoplates, copper sulfide (CuS), Ag@Au-CuS@lipo 53
4.2 Cell uptake and cytotoxicity of Ag@Au-CuS@lipo-Den 62
4.3 Penetration of Ag@Au-CuS@lipo-Den nanoparticles into tumor spheroids 66
4.4 In vivo animal experiment 68
4.4.1 In vivo animal experiment for biodistribution 68
4.4.2 The influence of immune system triggered by high-frequency magnetic field (HFMF) 71
4.4.3 In vivo therapy 73
Chapter 5 Conclusions 75
Reference 76
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