臺灣博碩士論文加值系統

開發一個簡單、便宜，對於癌症只需單次且小劑量之非侵入式檢
測及治療方法極具挑戰性。為使此治療診斷方式有效，具有雙模態成
像功能的單光誘導光熱及光動力試劑受到高度期待。癌症一直被認為
是對世界人口的巨大威脅，導致全球數百萬的威脅。一般來說，癌症
會弱化體內的免疫系統，使其更容易被細菌攻擊及繼發感染。細菌性
病原體一般來說已在我們的生活中造成更嚴重的損害，且缺乏有效的
常規抗生素使我們面臨重大的疫情威脅。由於對於病原體具有非傳統
的殺傷及抑制方式，基於奈米材料的現代治療診斷技術提供了各種解
決方案。對於癌症來說，儘管受到應用系統和遞送過程的限制，且通
常藥物被設計成對於其目標疾病具有專一性，最近關於藥物遞送的研
究已經顯示出進展。
請注意設計多功能的奈米材料是一項非傳統且具有挑戰性的任務。
我們主要關注於採用奈米材料在生物醫學與治療領域的潛在突破。現
代的治療診斷技術需要專門的運送系統，而其餘的因不溶於水而受到
限制。
被認為是奇蹟材料的石墨烯，因其優越的電性、熱穩定性、表面
積，光學特性、機械性質以及良好的導電性，已經成為一種有前景的
材料。除了這些有趣的特性之外，石墨烯也是一種具有生物相容性的
材料，不像其他如富勒烯和奈米碳管等材料（CNTs）。
在最近設計的奈米材料，有機污染物，微生物感染等盛行全球且
進一步的風險中，將染料傾倒在地下水中，導致污染水源是最明顯的。
未經處理且含有染料的工業廢水在我們的土地和水體上釋放，這需要
更具策略性和更有效的水處理方法。對於這樣的問題，我們設計了一
種基於石墨烯的奈米複合材料，用於在保持生物相容性的情況下，有
效地破壞染料及有機污染物，如羅丹明B 和細菌。我們設計的材料
不僅能成功的殺死癌細胞，還能殺死細菌性病原體。
因此，發現如RGOPAA，DHA @ MGPA-ICT 和GV-PEI 等材料，
我們提供了多種具有生醫診斷和成像、光動力和光熱研究、磁導藥物
遞送，及對於被有機廢棄物污染之廢水之處理，一個有效且廣泛的潛
在應用材料選擇。

Developing of a simple and cost-effective strategy to diagnose and
treat cancer with single and minimal dosage through noninvasive methods
are highly challenging. To make such theranostic strategy effective, single
light induced photothermal and photodynamic reagent with dual modal
imaging capability is highly desired. Cancer has long been considered a
huge threat to the world population, leading to millions of threat worldwide.
Often cancer leads to weaker immune system in the body, making it easier
for bacterial attacks and secondary infections. Bacterial pathogens have
caused much more havoc in our lives in general and the lack of effective
conventional antibiotics exposes us to a major epidemic threat. Modern
therapeutics based on nanomaterials has provided various solutions due to
their unconventional approach towards pathogen killing and inhibition.
For cancer, recent studies on drug delivery have shown progress although
limited by the system of application and delivery process and often, drugs
are designed very specific to their target disease.
Designing of a multifunctional nanomaterials, keeping in mind their
unconventional applications is a challenging task. Our major concern must
be to employ nanomaterials for a potential breakthrough in the field of
biomedicine and therapy. Modern therapeutics demand for specialized
delivery system while rest are limited due to their insolubility in water.
Graphene, considered a wonder material has emerged as a promising
material owing to its fascinating properties in superior electronics, thermal
stability, specific surface areas, optical and mechanical, as well as good
conductivity. Apart from the other intriguing properties, graphene is also a
biocompatible material, unlike its other counter parts like fullerene and
carbon nanotubes (CNTs).
Among the globally prevalent and further emanating risks of recently
engineered nanomaterials, organic pollutants, microbial infections, etc.,
dumping of dyes in ground water thereby contaminating the water supply
is the most notable. Untreated Industrial wastes containing dyes are
released on our grounds and water bodies, which demands for a more
strategic and efficient water treatment methods. For such concerning
problems, we have designed a graphene based nanocomposite for
effectively destroying dyes and organic pollutants, like Rhodamine B and
bacterial population while still maintaining its biocompatibility. Our
designed materials have not only been successful in killing cancer cells,
but also bacterial pathogens.
Therefore, the discovery of materials like RGOPAA, DHA@MGPAICT
and G-V-PEI, we provide a diverse choice of materials with effective
and vast potential applications in biomedical diagnostics and imaging,
photodynamic and photothermal studies, magnetically guided drug
delivery, as well as effective treatment for organic water pollutants.

Acknowledgement…………………………………………………………………..i
Abstract in Chinese………………………………………………………………..iv
Abstract in English………………………………………………………………..vi
List of Schemes…………………………………………………………………..xiii
List of Figures……………………………………………………………………xiv
List of Tables……………………………………………………………………xviii
List of Publications……………………………………………………………....xix
Chapter 1. Background and Introduction
1.1 Introduction………………………………………………………………...1
1.2 Gold Nanomaterials………………………………………………………..4
1.3 Graphene Nanomaterials………………………………………………….5
1.4 Quantum dots………………………………………………………………8
1.5 Carbon based and polymer materials…………………………………….9
1.6 Magnetic Nanomaterials……………………………………………….....10
1.7 References…………………………………………………………………11

Chapter 2. Exploring the photothermal hot spots of graphene in the first and second biological window to inactivate cancer cells and pathogens
2.1 Introduction…………………………………………………………………..21

2.2 Experimental Section
2.2.1 Synthesis of Graphene Oxide……………………………………………….23
2.2.2 Preparation of reduced graphene oxide (RGO)……………………………..23
2.2.3 Preparation of RGOPAA…………………………………………………....24
2.2.4 Cell culture procedures…………………………………………………….....25
2.2.5 Cellular uptake of RGOPAA using confocal laser scanning optical microscopy (CLSM)…………………………………………………………………………….25
2.2.6 Cell viability assay…………………………………………………………..25
2.2.7 Heat shock protein (HSP 70) expression analysis by flow cytometry………26
2.2.8 Bacterial culture……………………………………………………………..27
2.2.9 Photothermal tests for bacteria………………………………………………28
2.3 Results and discussion
2.3.1 Synthesis, characterization, and photothermal properties of RGOPAA…….29
2.3.2 Cellular uptake and photothermal therapeutic experiments of RGOPAA with HeLa cells………………………………………………………………………….32
2.3.3 Photothermal therapeutic experiments of RGOPAA with S. aureus bacteria..37
2.4 Conclusion………………………………………………………………….....40
2.5 References……………………………………………………………………..42

Chapter 3. Synergistic effect of Dihydroartemisinin Loaded Magnetic Graphene for PDT and PTT activity against B16 and HeLa cancer cells, and bacterial pathogens.
3.1 Introduction…………………………………………………………………...45
3.2. Experimental Section 3.2.1 Synthesis of GO…………………………………………………………….50
3.2.2 Synthesis of MRGO………………………………………………………...50
3.2.3 Preparation of MRGOPAA…………………………………………………51
3.2.4 Preparation of MGPA-ICT………………………………………………….52
3.2.5 Preparation of DHA@MGPA-ICT.................................................................52
3.2.6 Cell culture procedures (HeLa Cell Line)………………...............................53
3.2.7 Cell culture procedures (B16F1 Cell Line)…………………………………53
3.2.8 Cellular uptake of RGOPAA using confocal laser scanning optical microscopy (CLSM)………………………………………………………………………….…53
3.2.9 ROS generation study………………………………………………………..54
3.2.10 In vitro cell culture and cytotoxicity measurement………………………...55 3.2.11 In vitro PDT/PTT treatment………………………………………………..55
3.2.12 Bacterial culture…………………………………………………………....56
3.2.13 Photothermal tests for bacteria……………………………………………..57
3.2.14 In vivo toxicity tests using zebrafish embryo model……………………….57 3.2.15 In vivo embryonic toxicity test for zebrafish ……………………………....58
3. 3 Results and discussion
3.3.1 Synthesis, characterization, and photothermal properties of DHA@MGPA-ICT............................................................................................................................58
3.3.2 MRI Imaging to assess the Elemental composition…………………………61 3.3.2 Cellular uptake and PDT-PTT experiments of DHA@MRGOPAA-ICT with HeLa and B16F1 cells……………………………………………………………...64
3.3.3 Demonstration of ROS generation in HeLa cancer cells……………………68 3.3.4 Photothermal therapeutic experiments of DHA@MGPA-ICT with S. aureus bacteria and E.Coli………………………………………………………………...70
3.3.5 In vivo toxicity in zebrafish embryo using MRGOPAA and DHA@MGPA-ICT............................................................................................................................72
3. 4 Conclusion……………………………………………………………………75
3. 5 References…………………………………………………………………....76

Chapter 4. ROS Generating Vanadium Pentoxide Nanostructures wrapped in PEI functionalized Graphene for Photocatalytic and Theranostic Activity.
4.1 Introduction…………………………………………………………….……..81
4.2 Experimental Section
4.2.1 Synthesis of Vanadium Pentoxide…………………………………………...85
4.2.2 Synthesis of Vanadium Pentoxide nanostructures wrapped in PEI functionalized Graphene (G-V-PEI)……………………………………………… 86
4.2.3 Bacterial culture…………………………………………………………….. 86
4.2.4 1,3-Diphenyl isobenzofuran (DPBF) experiments………………………. 87
4.2.5 Photothermal tests for bacteria…………………………………………….... 88
4.2.6 Photocatalytic Activity of the G-V-PEI Nanocomposite………………….... 88 4.2.7 In vivo toxicity tests using zebrafish embryo model………………………...89 4.2.8 In vivo embryonic toxicity test for zebrafish……………………………….. 89
4. 3 Results and Discussion
4.3.1 Synthesis, Characterization and phototherapeutic properties of G-V-PEI nanocomposite………………………………………………………………….….90
4.3.2 Phototherapeutic experiments of G-V-PEI with S.aureus and E.coli……….95
4.3.3 Photocatalytic Degradation of Rhodamine B……………………………….97
4.3.4 In vivo toxicity in zebrafish embryo using pure V2O5 and G-V-PEI nanocomposite……………………………………………………….…………….99
4.4 Conclusion……………………………………………………………………101
4.5 References………………………………………………………………………..102

Chapter 5.
Summary…………………………………………………………………………106
References for All Chapters…………………………………………………….110
Appendix…………………………………………………………………..……..xxi

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Chapter 2
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