Multi-functional Graphene as a New Theranostic Agent in Biomedicine and an Effective Adsorbent for Safe Drinking Water

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研究生：	干尼 Ganesh Gollavelli
論文名稱：	Multi-functional Graphene as a New Theranostic Agent in Biomedicine and an Effective Adsorbent for Safe Drinking Water
指導教授：	凌永健 Ling, Yong-Chen
口試委員:	凌永健 Ling,Yong-Chien 黃賢達 Huang, Shang-Da 立袁林 Lin, Lih-Yuan 林針針 Lin, Pin-pin 麥富德 Mai, Fu-Der
學位類別：	博士 Doctor
系所名稱：	理學院 - 化學系 Department of Chemistry
論文出版年：	2012
畢業學年度：	100
語文別：	英文
論文頁數：	130
中文關鍵詞：	石墨，飲用水和生物醫學
外文關鍵詞：	Graphene, Drinking water and Biomedicine
相關次數：	點閱：1 下載：0
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Designing multi-functional nanomaterials with emerging properties is a challenging task. There is every concern to employ nanomaterials for the potential breakthrough in biomedical applications such as cellular imaging, diagnosis, and therapeutics. Modern therapeutics preferentially require some specialized delivery systems in order to maximize their therapeutic efficacy of water insoluble drugs, imaging them at cellular level and also to monitor or manipulate with an external magnetic force.
Graphene, a fascinating material has recently emerged with many intriguing properties including electrical, thermal, optical, sensing, high surface area and biocompatibility. The single or fewer layered structure of graphene provides richness for diversified surface chemistry on both sides of the sheet including edges. By considering the advantage of these properties, we synthesized multifunctional graphene (MFG) / smart magnetic graphene (SMG) with multi functions such as magnetic, water-dispersibility, and fluorescent properties using microwave-heated and sonication-assisted process within one minute. The MFG has been demonstrated as a potential fluorescent marker with lower cytotoxicity for both in vitro and in vivo imaging. Further, it was also non-covalently functionalized with near infrared light absorbing photo sensitizer (PS), SiNc4, to impart MFG-PS. The MFG-PS hybrids have shown excellent phototoxicities in killing the cancer cells through less invasive therapeutic treatments, such as, photodynamic and photothermal therapies. In the clinical diagnosis, MFG-PS is capable to locate the diseased area (fluorescence from MFG) as well as employed for photodiagnosis and molecular imaging, known as, photosensitizer fluorescence detection (PFD).
Among many globally pervasive and emerging risks such as, microbial infections, environmental impact of engineered nanomaterials (NM), and inorganic / organic contaminants, incessant supply of potable and safe drinking water is on the top notch. Many efforts have been driven to decontaminate the potable waters. The need for an efficient, cost-effective, robust, and handy technology for the decontamination of downstream water without endangering human health is still tremendous. We tackle these challenges by SMG in a single roof. The SMG possesses increased active adsorption sites with tunable superparamagnetic property, facilitating the adsorption and magnetic separation of aqueous Cr (VI), As (V) and Pb (II) with ~99% removal efficiencies down to 1 ppb level and decontaminant the organic pollutants. In addition to the efficient adsorption capability, SMG also exhibits better disinfecting action towards E. coli bacteria with 100% killing efficacy and low toxicity towards zebrafish without inducing any abnormalities.
Therefore, the discovery of MFG / SMG provides new opportunities in the biomedical field with diversified potential applications such as in biomedical diagnostics, magnetically guided drug / gene delivery, and photothermal / photodynamic therapies as well as in supplying the safe drinking water.

多功能納米材料的設計與新興物業，是一項艱鉅的任務。是每一個關注採用納米材料在醫學領域的應用，如細胞成像，診斷和治療的潛在突破。現代療法的優先需要一些專門的輸送系統，以最大限度地發揮其療效不溶於水的藥物，成像在細胞水平上，他們還監視或與外部的磁力操縱。
石墨，一個迷人的材料最近出現了許多有趣的性質，包括電學，熱學，光學，遙感，高比表面積和生物相容性。石墨的層狀結構的單一或較少提供多元化的表面化學在紙張的兩面，包括邊緣豐富。考慮到這些屬性的優勢，我們合成多功能石墨（MFG）/智能磁性石墨（SMG）的磁性，水分散性，熒光特性，如多在一分鐘之內使用微波加熱和超聲波輔助進程的功能。已被證明在體外和體內成像的細胞毒性較低作為一個潛在的熒光標記的工業。此外，它也是非共價與近紅外光吸收光敏劑（PS），SINC4，傳授MFG-聚苯乙烯功能。股份有限公司-PS的雜交已通過微創的治療方法，如光動力和光熱療法，在殺死癌細胞的優秀phototoxicities。在臨床診斷，MFG-PS是能夠找到病變區域（從工業熒光）以及就業photodiagnosis和分子成像，作為光敏劑，熒光檢測器（PFD）。
在眾多全球範圍內普及和新興的風險，如微生物感染，納米材料對環境的影響（新墨西哥州），無機/有機污染物，飲用水和安全的飲用水供應不斷，是頂尖的。帶動了許多努力來淨化飲水。下游水不危害人體健康的去污效率，成本效益，穩健，和方便的技術的需求仍然是巨大的。我們面對這些挑戰，通過在一個單一的屋頂衝鋒槍。 SMG的具有增加主動可調的超順磁性吸附位，促進水溶液中Cr（VI）的吸附和磁分離，（五），鉛（二）〜99％的去除率下降到1 ppb級和去污劑中的有機污染物。除了高效的吸附能力，SMG也表現出較好的消毒對大腸桿菌的行動，殺害100％，沒有引發任何異常的療效和毒性低，對斑馬魚。
因此，發現了MFG /衝鋒槍在生物醫學領域提供了新的機遇與多元化的應用潛力，如在生物醫學診斷，藥物/基因傳遞磁導，光熱/光動力療法，以及提供安全飲用水。

Table of Contents

Chapter-1] Background and Introduction
1 Background    1
2 Quantum dots    2
3 Gold Nanomaterials    3
4 Iron oxide nanoparticles    4
5 Carbon nanomaterials    6
Chapter-2] Multi-functional graphene as an in vitro and in vivo imaging probe
1 Background and Introduction    12
2 Experimental Section    13
2.1 Synthesis of graphene oxide (GO)    13
2.2 Synthesis of magnetic graphene (MG)    14
2.3 Synthesis of multi-functional graphene (MFG)    15
2.4 Cell Cultures    15
2.5 MTT Assay    16
2.6 Lactate Dehydrogenase Release (LDH) Assay    16
2.7 Reactive Oxygen Species (ROS) Assay    17
2.8 Annexin V Apoptosis Assay    17
2.9 CLSM measurement    17
2.10Microinjection of MFG into zebrafish embryos and microscopic measurements    18

3 Results and Discussion    19
3.1 Synthesis and characterization of MFG    19
3.2 In vitro cytotoxicity    26
3.3 In vitro imaging    32
3.4 In vivo toxicity in zebrafish    35
3.5 In vivo imaging in zebrafish    39
4 Conclusions    40
5 References    41
Chapter-3] Magnetic and Fluorescent Graphene based Theranostic Nanoprobes for Multimodal Imaging and Photothermal / Photodynamic Destruction of Cancer Cells 
1 Introduction    51
2 Experimental Section    54
2.1 Synthesis of graphene oxide (GO)    54
2.2 Synthesis of magnetic graphene (MG)    54
2.3 Synthesis of magnetic and fluorescent graphene (MFG)    55
2.4 Synthesis of MFG-PS hybrids    56
2.5 Cell cultures    56
2.6 Dark and phototoxicities by MTT assay    56
2.7 LDH assay    57
2.8 CLSM measurements    58
2.9 SOSG experiments    58
2.10 ROS assay    58
2.11 Annexin V apoptosis assay    59
2.8 Changes in mitochondrial membrane potential    59
2.9 Heat shock protein expression analysis    60
3 Results and Discussion    60
3.1 Synthesis and characterization of MFG-PS    60
3.2 Co-localization of MFG-PS in HeLa cells using CLSM    64
3.3 MRI imaging of MFG    65
3.4 Dark and phototoxicities of MFG    67
3.5 Cellular events responsible for phototoxic MFG-PS mediated cellular death    68
4 Conclusions    73
5 References    74
Chapter-4] Smart magnetic graphene as a disinfectant, biocompatible and heavy metal adsorbent for safe drinking water
1 Introduction    80
2 Experimental Section    82
2.1 Materials    82
2.2 Preparation of SMG by solid state microwave induced process    82
2.3 Characterization    83
2.4 Disinfecting study    84
2.5 Confocal microscopy    84
2.6 Zebrafish biocompatibility    85
2.7 Adsorption of Cr (VI), As (V) and Pb (II)    85

3 Results and Discussion    86
3.1 Preparation and characterization of SMG    86
3.2 Disinfecting action of SMG against E.coli    90
3.3 In vivo zebrafish biocompatibility    93
3.4 Adsorption of HM    95
4 Conclusions    99
5 References    100
Chapter-5] Comparitive studies of graphene oxide and magnetic graphene on Bisphenol-A adsorption, antibacterial activity and in vitro toxicity
1 Introduction    110
2 Experimental Section    111
2.1 Materials    111
2.2 Preparation of SMG by solid state microwave induced process    111
2.3 Characterization    113
2.4 Adsoprtion of BPA by MG    113
2.5 MTT assay    114
2.6 LDH assay    115

2.4 Results and Discussion    115
3 Conclusions    124
4 References    124

Summary and Perspectives…………………………………………………….127

                                

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