董成淵臺灣大學：物理研究所駱文Lo, WenWenLo2007-11-262018-06-282007-11-262018-06-282004http://ntur.lib.ntu.edu.tw//handle/246246/54480雙光子顯微術 (Two-photon microscopy)具有非侵入性檢測、重構三維影像能力並且可以深入生物組織的特點，近年來已廣泛應用於各種生醫研究中，在我的研究中，我利用雙光子顯微術的這種特點研究透過皮膚藥物傳遞的路徑與機制。 透過皮膚遞送藥物 (Transdermal drug delivery) 有著許多優點，這種非侵入性的藥物傳遞方式有著數種優於口服或靜脈注射之處，諸如：避免藥物在經過消化道遭受破壞、維持血液中藥物濃度穩定、較易應用於較長期的療程、減低藥物用量，甚至還有增加病患配合度之效。關於透過皮膚的藥物傳遞，雖然已有許多不同的機制模型與研究，關於傳遞效率增益的研究更是不計其數，縱使有許多實驗間接證實利用一些物理性或化學性的手段，增加了藥物傳遞的效率，然而我們對這些增益法是否或是如何其傳遞之路徑與機制的了解仍然有限，關於藥物分子大小在藥物傳遞中所占地位亦一無所知。透過雙光子顯微術，我們將有機會直接監測透過皮膚傳遞所選定的不同大小分子的傳遞路徑，並且研究利用油酸分子增加藥物穿透皮膚效率的機制。 研究中用於模擬藥物的類分子－量子點 (Quantum dots)－ 是由半導體材料所構成的奈米粒子，而這種大小尺度與許多重要的生物巨分子十分相似。近年來所研發出的藥物有很大一部分是蛋白質分子或胜肽分子，透過皮膚將這些藥物送入病患體內可能是較佳的作法，利用量子點與這些巨分子大小的相似性，同時我們可以修飾量子點的表面化學使其可與特定的組織或細胞結合，因此運用量子點作為模擬透過皮膚所傳遞的藥物，將不失為一種研究藥物傳遞之路徑與機制的好方法。此外，量子點在光學性質上具有數種優於傳統螢光染劑的特點，如：發射光譜較窄、可調變發射光譜、較穩定的光化學性質、較長的半生期。若能透過皮膚將之傳送入生物體內，將可大幅增進生物醫學影像的品質。 基於上述的原因，我的論文內容將比較水溶性量子點與水溶性螢光分子在皮膚中傳遞的路徑差異，以及油酸在傳遞過程中影響量子點或螢光分子的方式。Luminescent quantum dots (QDs) are attractive fluorescent probes in bioimaging application. Broad excitation spectrum, narrow and tunable emission spectrum, high photochemical stability and chemical modifiability are among the desirable properties of QDs. In this work, we investigate the feasibility of administering QDs (7 nm in size) through transdermal delivery pathway using multiphoton fluorescence imaging. Our results show that QDs of this size range can be effectively delivered through intercellular pathways using the chemical enhancer oleic acid. This work has implications for understanding the transport of important biological macromolecules. Our experiments infer two important results. First QDs can hardly transport into skin without the chemical enhancer oleic acid. Furthermore, QDs deliver primarily through intercellular pathways. In contrast, fluoresein molecules deliver through both intercellular and intracellular region. Our results imply that the size of nanoparticles may dominate the nature of transdermal transport mechanism and in the future, we plan to investigate the mechanism by manipulating the size of nanoparticles and their surface chemistry.UTILIZING MULTIPHOTON MICROSCOPY TO MONITOR CHEMICALLY ENHANCED TRANSDERMAL DELIVERY PATHWAYS OF LUMINESCENT QUANTUM DOTS 1 CHAPTER 1 INTRODUCTION 4 1-1 Advantages of transdermal drug delivery and enhanced methodologies 4 1-2 Transdermal delivery of quantum dots 5 1-3 Introduction to multiphoton microscopy 7 CHAPTER 2 SKIN AND TRANSDERMAL DELIVERY 9 2-1 The structure of skin19 9 2-2 Several possible delivery pathway20-22 12 CHAPTER 3 QUANTUM DOTS AS BIOLOGICAL LABELS 15 3-1 Basic physics of quantum dots 15 3-2 Photostability and surface chemistry 17 CHAPTER 4 MULTIPHOTON MICROSCOPY 19 4-1 The fundamentals of two-photon excitation 20 4-2 The characteristics and the advantages of multiphoton microscopy 25 CHAPTER 5 EXPERIMENTAL SETUP 28 5-1 The setup of multiphoton microscope 28 5-2 Fluorescent dyes and quantum dots 30 5-3 Delivery methods 33 CHAPTER 6 RESULTS AND DATA ANALYSIS 35 CHAPTER 7 CONCLUSION 41 CHAPTER 8 ACKNOWLEDGEMENT 42 CHAPTER 9 REFERENCE 441475478 bytesapplication/pdfen-US量子點雙光子皮膚脈衝雷射顯微術油酸藥物傳遞multiphotonskinchemically enhancedquantum dotsmicroscopytwo-photonoleic acidtransdermal deliveryti-sapphire laserthesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/54480/1/ntu-93-R92222031-1.pdf