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研究生: 劉耀文
Bruce Yao-Wen Liu
論文名稱: 以金奈米棒之光熱效應破壞癌細胞之活體 研究
In vitro and in vivo photothermal destruction of tumors by gold nanorods and laser irradiation
指導教授: 陳洋元
Chen, Yang-Yuan
賈至達
Chih-TaChia
學位類別: 碩士
Master
系所名稱: 物理學系
Department of Physics
論文出版年: 2012
畢業學年度: 100
語文別: 英文
論文頁數: 75
中文關鍵詞: 光熱治療表面電漿子共振金奈米棒
英文關鍵詞: Photothermal therapy, Surface plasmon resonance, gold nano rods
論文種類: 學術論文
相關次數: 點閱:145下載:6
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  • 因為金奈米棒可吸收近紅外光以及表面電漿共振之特性,讓金奈米棒具有很棒的光熱轉換效率,此高效率產生之熱量可以有效的殺死癌細胞。藉由一步合成法,可以有效率的合成出吸收峰值落於 800 nm 的金奈米棒,藉由紫外光-可見光光譜儀、穿透式電子顯微鏡、Zeta Potential以及ICP-MS測量儀之檢測,可以清楚了解金奈米棒的特性以及江奈米棒定量。在細胞以及活體實驗上均可觀察到利用金奈米棒所進行之光熱作用在治療上的效果;在細胞實驗上,藉由染劑的觀察,可以有效判斷出癌細胞的存活狀態,實驗結果大致可得到以下結論,金奈米棒被細胞吞噬之數量以及雷射之能量為影響細胞存活的最大變因;在活體實驗中,由於選用老鼠耳朵當做實驗對象,因此癌細胞增長所導致的血管新生現象也可以很容易就被觀察到,活體實驗大致流程為下:將癌細胞植入老鼠左右耳上,約到第十天左右,腫瘤體積約為30 mm3 ,此時將 ~10 μg金奈米棒注射於腫瘤處,最後進行雷射之光熱治療。從組織切片之結果以及腫瘤體積量測之生長曲線結果判斷,可以觀測到金奈米棒受雷射激發後所產生之光熱效應,可以嚴重的破壞癌細胞的增長甚至是將腫瘤的大部分患部移除。這些結果可以看出光熱治療此機制在未來臨床應用上具有很大的潛力。

    Gold nanorods (AuNRs) have tunable plasmon-resonant absorption in near-infrared (NIR) region and excellent photothermal transduction, making them attractive for the next generation of cancer therapy. In this study, AuNRs with a longitudinal absorption peak at 800 nm were prepared by one-pot synthesis method, and are further characterized by absorption spectra, electron microscopy, zeta potential and inductively coupled plasma mass spectroscopy. Both in vitro and in vivo approaches were performed to evaluate AuNRs’ therapeutic efficacy. According to dual color fluorescent staining imaging, cell destruction greatly depends on influence of energy and the amount of nanorods taken up. Laser-induced heating was applied to investigate the therapeutic effect of AuNRs, where the alteration in tumor size and vascular development was examined in the mouse ear. In the experiments, murine breast tumor cells were first introduced into the ears of Balb/c female mice. As the tumors grew to about 30 mm3 in size at the 10th day, AuNRs (~10 μg) were intratumorally injected into the ears directly, followed by employment of laser irradiation. The results of histological section and size measurements indicated that the photothermal effects of AuNRs are significantly destructive to the tumor development and may lead to final removal of the solid tumor. These results suggested that AuNRs have great potential in future clinical phototherapeutic applications.

    Abstract 中文摘要 Acknowledgements Table of contents i List of figures iii Chapter 1. Introduction 1 1.1 Properties of AuNRs 2 1.1.1 Surface plasmon resonance 3 1.1.2 Quantum confinement effect 4 1.2 Introduction of cancer 6 1.2.1 Differences between cancer and normal cells 7 1.2.2 Metastasis of cancer cells 9 1.3 Cancer therapies 11 Chapter 2. Experimental details 18 2.1 Chemicals 18 2.2 Instruments 20 2.3 Preparation of AuNRs 21 2.3.1 Synthesis of AuNRs 21 2.3.2 Surface modification with polystyrenesulfonate 24 2.3.3 Characterizations of AuNRs 26 2.3.3.1 UV-visible spectroscopy 26 2.3.3.2 Transmission electron microscope 28 2.3.3.3 Zeta potential measurement 29 2.3.3.4 Inductively coupled plasma mass spectroscopy 32 2.4 In vitro experiments 33 2.4.1 Cell line 33 2.4.2 Cell culture 34 2.4.3 Cell viability assay (MTT Assay) 35 2.4.4 In vitro photothermal therapy 37 2.4.4.1 Introduction to fluorescent Dyes 38 2.4.4.2 Confocal microscope and two-photon luminescence imaging 40 2.5 In vivo experiments 44 2.5.1 Animal model 44 2.5.2 Anesthesia 46 2.5.3 In vivo photothermal therapy 46 2.5.3.1 Experimental processes 46 2.5.3.2 Microscope system 48 2.5.3.3 Estimation of tumor size 48 2.5.3.4 Histological sections 49 Chapter 3. Results and discussions 50 3.1 Characterizations of gold nanorods 50 3.2 In vitro cancer cell photothermolysis mediated by AuNRs 56 3.3 In vivo tumor photothermal therapy 63 3.3.1 Power density test with the ears model 63 3.3.2 Tumor growth curve 64 3.3.3 Histological study 69 Chapter 4. Conclusions 71 References 73

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