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開發功能化微脂體平台於癌症熱治療

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Table of contentsCONTENTS PAGERecommendation letter from thesis advisor……………………..………Thesis/Dissertation oral defense committee certificate……………Acknowledgement iiiChinese abstract viEnglish abstract viiiT

able of contents xiList of figures xviiList of tables xxAbbreviations xxiChapter 1: Overview of Caner Thermal Therapies 11 Introduction 12 Background of thermal therapies 63 Objective 7Chapter 2: Applications of Magnetic Liposomes in Cancer Therapies 91 Introd

uction 91.1. MNPs and liposomes in cancer treatment 101.1.1. Significance of MNPs in cancer therapy 101.1.2. Significance of liposomes in cancer therapy 141.2. Preparation methods of MNPs, liposomes and magnetic liposomes 161.2.1. Preparation methods of MNPs 161.2.1.

1. Physical method 161.2.1.2. Biological method 171.2.1.3. Chemical method 171.2.2. Preparation methods of liposomes 181.2.2.1. Thin film hydration or Bangham method 191.2.2.2. Extrusion method 201.2.2.3. Reverse phase evaporation method 211.2.2.4. Superc

ritical reverse phase evaporation method 211.2.2.5. Detergent depletion method 221.2.2.6. Injection method 231.2.2.7. Microfluidic channel method 241.2.3. Preparation of magnetic liposomes (MLs) 252 Magnetic liposomes in cancer therapies 262.1. MLs for drug delive

ry and thermo-chemotherapy 262.2. MLs for gene delivery and combined gene therapies 312.2.1. MLs for gene delivery 322.2.2. MLs for combined gene therapies 332.3. MLs in photothermal/photodynamic therapy or magneto-phototherapy 342.3.1. Advantages of MLs for targeted ph

otothermal/photodynamic therapy …………………………………………………………………………362.3.2. Use of MLs in photothermal-AMF combined method (magneto-phototherapy) 372.4. Application of MNPs and MLs for cancer imaging and therapy 413 Conclusion 44Chapter 3: Optimization of the Preparation of Magnetic Li

posomes for the Combination Use of Magnetic Hyperthermia and Photothermia in Dual Magneto-Photothermal Cancer Therapy 471 Introduction 472 Materials and Methods 512.1. Materials 512.2. Synthesis of Citric Acid-Coated Iron Oxide Magnetic Nanoparticles (CMNPs) 522.3.

Preparation of Magnetic Liposomes (MLs) 522.4. Experimental Design 532.5. Charactrization of Physico-Chemical Properties 552.6. Heating Efficiency Induced by AMF and/or NIR Laser 562.7. Intracellular Uptake of MLs by Cancer Cells 572.8. In-vitro Biocompatibility of ML

s 592.9. In-vitro Cancer Cell Killing by AMF and/or NIR Laser 592.10. Flow Cytometry Analysis for Apoptosis/Necrosis 602.11. Statistical Analysis 613 Results and Discussion 613.1. Model Development and Optimization 613.2. Characterization of Physico-Chemical Prope

rties 683.3. Heating Efficiency Induced by AMF and/or NIR Laser 783.4. Intracellular Uptake of MLs 803.5. Thermally Induced Cancer Cell Killing In-vitro 824 Conclusion 86Chapter 4: Dual Targeted Magnetic Photosensitive Liposomes for Photothermal/Photodynamic Tumor Therapy

871 Introduction 872 Materials and Methods 902.1. Materials 902.2. Synthesis of citric-acid coated iron-oxide magnetic nanoparticles 912.3. Synthesis of HA-PEG 922.4. Preparation of liposomes 922.5. Determination of encapsulation efficiency of CMNPs and ICG

932.6. Characterization of HA-PEG-MPLs 942.7. Temperature elevation induced by NIR laser irradiation 952.8. In-vitro cell culture experiments 952.9. In-vivo antitumor efficacy 962.10. In-vivo IVIS imaging 982.11. Statistical Analyses 993 Results and Discuss

ion 993.1. Characterization of HA-PEG-MPLs 993.2. In-vitro photothermal effects of HA-PEG-MPLs 1053.3. In-vitro cytotoxicity of HA-PEG-MPLs 1063.4. In-vivo effects of HA-PEG-MPLs 1083.5. In-vivo antitumor and tumor targeting effects from IVIS imaging 1114 Conclusi

on 115Chapter 5: Concurrent Photothermal and Photodynamic Therapy of Intracranial Brain Tumor Xenografts with Convection Enhanced Delivery of Liposomal IR-780 1161 Introduction 1162 Materials and methods 1202.1. Materials 1202.2. Preparation of IR-780 loaded liposomes 1

202.3. Characteristic of IR-780 loaded liposomes (ILs) 1212.4. Photothermal and Photodynamic effects study 1222.5. In vitro cell culture experiments 1232.6. Tumor cell implantation in xenograft mice brain 1252.7. Convection enhanced delivery 1272.8. In vivo temperatu

re measurements during NIR irradiation 1282.9. In vivo anti-tumor efficacy 1292.10. MRI and PET/CT study 1292.11. Bio-distribution 1302.12. Histology studies of tumor tissue 1312.13. Statistical analysis 1323 Results and discussion 1323.1. Characterization o

f ILs 1323.2. In vitro photothermal and photodynamic study 1383.3. In vitro cells experiments 1453.4. In vivo biodistribution 1483.5. In vivo photothermal effects 1503.6. Anti-tumor efficiency 1523.7. MRI and PET-CT studies 1553.8. Immunohistochemical analys

is 1594 Conclusion 163Chapter 6: Conclusions and Outlooks 1641 Summary 1642 Future perspective 165REFERENCES 166List of figuresFigure 2.1 Schematic diagram of MNPs or MLs induced with AMF. 12Figure 2.2 Schematic representations of different kinds of surface modified lip

osomes. 15Figure 2.3 The drug release mechanism from TSMLs 28Figure 2.4 The hyperthermia modality in magneto-phototherapy with MLs induced by MHT with AMF treatment, laser treatment or dual MHT/laser treatments. 38Figure 3.1 The Pareto charts of EE and Size. 65Figure 3.2 Predicted v/s Ob

served value Plots. 66Figure 3.3 Response surface Contour 3D plots. 67Figure 3.4 Particle size and surface charge distribution from DLS and TEM images. 70Figure 3.5 Magnetic liposomes stability measurements with NTA. 72Figure 3.6 XRD, FTIR, SQUID and TGA analysis of CMNP and MLs. 75F

igure 3.7 In vitro heating efficiency of CMNPs and MLs as induced by magnetic hyperthermia (MH) and/or photothermia (PT). 76Figure 3.8 Particle uptake studies with U87 cancer cells. 82Figure 3.9 In-vitro cells biocompatibility and cytotoxicity measurements. 83Figure 3.10 Flowcytometry analy

sis of MLs with different treatments 85Figure 4.1 Schematic illustration of HA-PEG-MPLs for dual targeted photothermal or photodynamic cancer therapy. 90Figure 4.2 Liposomes size from DLS and Cryo-TEM 100Figure 4.3 Characterization of different samples by XRD and FTIR. 102Figure 4.5 The

ex vivo photothermal effects of different samples 105Figure 4.6. The in vitro cell cytotoxicity and live/dead cell assays. 107Figure 4.7 In vivo photothermal effects. 110Figure 4.8 Representative photographs of the tumor-bearing mice 110Figure 4.9 The tumor volume, body weight and surviv

al curve of different groups. 112Figure 4.10 H&E and immunohistochemical analysis in tumor site 112Figure 4.11 The in vivo bioluminescence and fluorescence imaging by IVIS 114Figure 5.1 CED infusion cannulas and their parts 126Figure 5.2 Demonstration mice receiving samples via CED metho

d 127Figure 5.3 schematic diagram of ILs and their characterization 133Figure 5.4 UV-visible and FTIR spectroscopy 134Figure 5.5 Photothermal stability of free IR-780 and ILs 135Figure 5.6 Stability of ILs in FBS measured from nanoparticle tracking analysis. 136Figure 5.7 In-vitro pho

tothermal changes with NIR laser irradiations 139Figure 5.8 Photothermal stability of ILs and Free IR-780 140Figure 5.9 ROS generation detected by UV-visible. 143Figure 5.10 Cell cytotoxicity measurements with MTT and from flow cytometry. 144Figure 5.11 Particle uptake studies with confo

cal laser scanning microscopy. 146Figure 5.12 The bio-distribution analysis of ILs via CED. 149Figure 5.13 in-vivo photothermal effects. 151Figure 5.14 The antitumor efficiency by IVIS, the body weight and survival. 153Figure 5.15 Magnetic resonance images and tumor volume 156Figure 5

.16 PET-CT molecular imaging analysis 158Figure 5.17 H&E and immunohistochemical staining 160Figure 5.18 H&E staining of different organs of mice of all three groups. 161List of tablesTable 2.1 Examples of preparation of magnetic liposomes 25Table 3.1 The central composite design showing

the independent variables and levels used in the experiments 54Table 3.2 Central composite design arrangement and observed responses. 62Table 3.3 Validation of the model with predicated experimental values 65Table 3.4 Size and zeta potentials values. 68Table 3.5 Specific absorption rate

s (SARs) of CMNPs and MLs at 0.6 mg/mL CMNP equivalent1. 73Table 3.6 Apoptotic and necrotic analysis form flow cytometry analysis. 79Table 4.1 Particle size and zeta potential of CMNPs, MPLs and HA-PEG-MPLs. 99Table 5.1 Size and zeta potential values of ILs 129Table 5.2 Survival times of

mice treated in different groups 155Table 5.3 The standardized uptake values (SUVmax) of Ga68-RGD and Ga68-FAPI 159Table 5.4 Hematological parameters and biochemistry analysis in different treatment groups. 162