Hybrid Core-Shell Micro-Nanospheres for Sensing, Bioimaging and Photothermal Therapy Applications

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研究生：	KARTHIKEYAN MANIVANNAN KARTHIKEYAN MANIVANNAN
論文名稱：	Hybrid Core-Shell Micro-Nanospheres for Sensing, Bioimaging and Photothermal Therapy Applications Hybrid Core-Shell Micro-Nanospheres for Sensing, Bioimaging and Photothermal Therapy Applications
指導教授：	陳建光 Jem-Kun Chen
口試委員:	蔡協致 Hsieh-Chih Tsai 鄭智嘉 Chih-Chia Cheng 許蕙玲 Hui-Ling Hsu 游佳欣 Jia Youxin 李愛薇 Ai-Wei Lee 黃啟賢 Chi-Hsien Huang 陳建光 Jem-Kun Chen
學位類別：	博士 Doctor
系所名稱：	工程學院 - 材料科學與工程系 Department of Materials Science and Engineering
論文出版年：	2018
畢業學年度：	106
語文別：	英文
論文頁數：	149
中文關鍵詞：	核-殼、二氧化矽微球、型態結構色、光子晶體、聚合、生物感測器、生物影像、光熱治療
外文關鍵詞：	Core-shell, SiO2 microsphere, Structural color, Photonic crystals, PNIPAM@SiO2 microspheres, Biosensors, Ag@SiO2@Agseed, Ag@SiO2@AgNPs, Nanosphere
相關次數：	點閱：302 下載：0
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查詢本校圖書館目錄查詢臺灣博碩士論文知識加值系統勘誤回報

現今核-殼材料成為研究人員所關注的材料之一，是因為在尺寸及形狀的可控性。
核-殼結構存在有像是球狀、星狀、螺旋狀、偏心圓、管狀等等，但以球狀最為廣泛應用，
其中包含光子晶體、感測器、生物影像及光熱療法，是由於核-殼結構在膠體狀態中有好
的遷移率及好的分散性質，高的堆積密度和表面具有低的光散射性質。本篇論文以球狀核
-殼材料二氧化矽-聚(N-異丙基丙烯醯胺) (PNIPAM)和 Ag@SiO2@Ag 奈米粒子為基礎進行
三個研究方向及應用。
第一部分為利用均一尺寸的二氧化矽微球和微米矽球接枝上熱響應的 PNIPAM 高
分子刷並探討其光子晶體行為。藉由表面原子自由基轉移聚合法 (ATRP)，利用
CuBr1/CuBr2/1,1,4,7,7-pentamethyldiethylenetriamine (PMDETA)作為催化系統，將異丙基丙
烯醯胺包覆在矽球表面。在矽球表面接枝的厚度由 FTIR、TGA、SEM、TEM 鑑定。
SiO2@PNIPAM 核-殼材料會和周圍的微米矽球會形成如光子晶體般緊密堆積。此外，樣
品通過用綠色激光指示器處理來評估。
在第二部份我們著重於利用無機矽球表面接枝 poly(N-isopropylacrylamide) (PNIPAM)
有機高分子刷之核-殼結構。先利用表面起始之原子自由基轉移聚合法將異丙基丙烯醯胺
聚合於矽球表面我們先將異丙基丙烯醯胺、甲醇和純水以一定比例混和，再加入含有
CuBr1/CuBr2/1,1,4,7,7-pentamethyldiethylenetriamine (PMDETA)之催化系統中，以此得到表
面接枝 NIPAM 高分子刷的微米矽球。所得到之 PNIPAM@SiO2 微球會再以 FTIR, TGA,
XPS, SEM, TEM 等儀器分析其性質。固定化血紅蛋白（ Hb）具有生物電活性，並且在電
化學實驗中藉由經修飾之電極測定固定化血紅蛋白時，觀察到一個明確的-0.38V 電位的
氧化還原峰。經修飾電極展現出很廣的線性檢測範圍，從 0.1μM 到 333μM，限制極限為
0.07μM。因此，修飾電極在檢測生物電活性中的 H2O2與以酶為基材的生物傳感器應用中
具有很高的潛在應用價值。
在第三部分裡，我們做出了 Ag@SiO2@Agseed 和 Ag@SiO2@AgNPs核-殼粒子應用在
生物顯影與光熱治療癌症上，我們使用了較環保合成的方式合成兩種不同的奈米複合粒子，
利用表面改質技術將矽殼層包覆在奈米銀表面粒子上控制其晶核成長。我們也使用了
ii
NaBH4 來當做還原劑並包覆在表面奈米銀複合粒子的銀奈米粒子上。我們對核-殼粒子做
了定型分析如：UV、PL、SEM、TEM 與 STEM 做表面型態的分析。可以藉由 UV 吸收特
定可見光波長的特性應用在生醫領域與光熱治療上，進而有效率的造成較大細胞如海拉細
胞凋亡達到治療的目的。在現今的研究中，我們用體外實驗研究發現了特有的多樣性與生
物相容性應用在癌症光熱治療與生物顯影，我們相信這樣的奈米材料未來在癌症治療上有
良好的前景

This PhD research mainly focused on the spherical-shape core-shell SiO2-poly(N
isopropylacrylamide) (PNIPAM) and Ag@SiO2@Ag micro-nanosphere hybrid core-shell micronanospheres for sensing, biomedical applications due to its attracting physicochemical property.
due to high mobility and good dispersion properties in paste state, high packing density and low
light scattering, high surface area, good biocompatibility.These doctoral studies divided in to
three applications such as photonic crystals (PCs), sensor and bioimaging (BI), Photothermal
therapy (PTT). From our results, we validate that.
1. In summary, a core-shell structure comprising a silica microsphere core and a
PNIPAM grafted shell. Because of their micro scale, the real-time packing behavior of
these particles in solution could be observed by OM. The visual red-to-blue color
change confirmed that the ordered packing of the microspheres in the water layer was
retained during the temperature increase. Hence, the synthesized PNIPAM-grafted
silica microspheres could provide an understanding of the packing mechanism of PCs
in water and could be used in a wide range of visual color sensor and optical
application.
2. we developed a simple method to prepare PNIPAM@SiO2 composite. The bioenzyme,
hemoglobin was immobilized on the surfaces of PNIPAM@SiO2 composite to fabricate
the Hb/PNIPAM@SiO2 composite electrode. The Hb exhibits direct electron transfer
with the modified electrode surface. Moreover, the Hb/PNIPAM@SiO2 modified
electrode detected H2O2 over wide well-linear concentration range. We believe that the
fabricated biosensor has great potential for biological and biomedical applications.
3. In summary, we have demonstrated the Ag@SiO2@Agseed and Ag@SiO2@AgNPs coreshell nanoparticles with enhanced therapeutic performance using a bioimaging and
excellent biocompatibility. The in vitro study shows the effective photothermal
destruction of HeLa cancer cells, we detected the large cells damage upon the light
irradiation Cell apoptosis imaging. We believe both materials are showed good
photothermal heat generation. Therefore, these nanoparticles would play effective
photothermal therapy agents play important roles in biomedical and the presented
research use as a promising therapeutic system for cancer.
Finally, that the the spherical-shape core-shell SiO2-poly(N isopropylacrylamide) (PNIPAM) and
Ag@SiO2@Ag micro-nanosphere hybrid core-shell micro-nanospheres for sensing, biomedical
applications. In detail, tailoring methods offer a multifunctional property in a single system,
which is going to needy in the near future. In addition, recently lot of researcher focused on this
novel material to resolve some of the longtime problem in the sensing promising therapeutic
system for cancer.

摘要--------------------------------------------------------------------------------------------------------------i
Abstract--------------------------------------------------------------------------------------------------------iii
Acknowledgement--------------------------------------------------------------------------------------------xi
List of Figures-----------------------------------------------------------------------------------------------xiv
List of schemes ---------------------------------------------------------------------------------------------xix
List of tables-------------------------------------------------------------------------------------------------xix
List of abbreviations----------------------------------------------------------------------------------------xx
Chapter 1: Introduction----------------------------------------------------------------------------------1
1. Silicon dioxide (SiO2)-------------------------------------------------------------------------------1
2. Silicon dioxide nanosphere and microsphere (SiO2)-------------------------------------------1
3. Synthesis (SiO2)-------------------------------------------------------------------------------------3
4. Core-shell (inorganic-polymer)--------------------------------------------------------------------4
5. Silver (Ag)--------------------------------------------------------------------------------------------4
5.1. Silver nanomaterial their applications ---------------------------------------------------5
6. Core-shell (inorganic-inorganic)------------------------------------------------------------------7
7. Eco- friendly polymer-------------------------------------------------------------------------------8
x
7.1. Thermoresponsive polymers --------------------------------------------------------------9
7.2. poly(N-isopropylacryamide)(PNIPAM)------------------------------------------------10
8. Atom Transfer Radical Polymerization (ATRP)----------------------------------------------12
9. Photonic crystal (PCs)-----------------------------------------------------------------------------13
9.1. One dimensional photonic crystals (1DPCs)-------------------------------------------14
9.2. Two dimensional photonic crystals (2DPCs)------------------------------------------14
9.3. Three dimensional photonic crystals (3DPCs)-----------------------------------------14
9.4. Bragg’s Law--------------------------------------------------------------------------------15
10. Biosensor------------------------------------------------------------------------------------16
11. Bioimaging---------------------------------------------------------------------------------18
11.1. Fluorescence Imaging --------------------------------------------------------------------18
12. Phototherapy--------------------------------------------------------------------------------20
12.1. Photothermal therapy (PTT)--------------------------------------------------------------20
Chapter 2: Research objectives -----------------------------------------------------------------------23
Chapter 3: Real-Time Packing Behavior of Core-Shell Silica@Poly(N-isopropylacrylamide)
Microspheres as Photonic Crystals for Visualizing in Thermal Sensing.------------------------24
1. Introduction-----------------------------------------------------------------------------------------25
2. Experimental Section------------------------------------------------------------------------------28
xi
2.1. Materials------------------------------------------------------------------------------------28
2.2. Synthesis of PNIPAM-Functionalized Silica Microparticle-------------------------28
2.3. Characterization----------------------------------------------------------------------------30
3. Results and Discussion----------------------------------------------------------------------------30
4. Conclusions-----------------------------------------------------------------------------------------43
Chapter 4: Facile synthesis of poly (N-isopropylacrylamide) coated SiO2 core-shell
microspheres via surface-initiated atom transfer radical polymerization for H2O2 biosensor
applications ----------------------------------------------------------------------------------------------44
1. Introduction------------------------------------------------------------------------------------45
2. Experimental section ------------------------------------------------------------------------47
2.1. Materials -------------------------------------------------------------------------47
2.2. Synthesis of NH2-functionalized SiO2 microspheres (NH2-SiO2)--------48
2.3. Synthesis of 2-bromoisobutyrate-functionalized SiO2 microspheres (BrSiO2) -----------------------------------------------------------------------------49
2.4. Synthesis of PNIPAM@SiO2 core-shell microspheres via ATRP-------49
2.5. Characterization-----------------------------------------------------------------50
2.6. Electrode preparation-----------------------------------------------------------50
3. Results and discussion-----------------------------------------------------------------------51
xii
3.1. Characterization of PNIPAM@SiO2 core-shell microspheres -----------51
3.2. Electrochemical behavior of Hb/NIPAM@SiO2 electrode----------------57
3.3. Real sample analysis------------------------------------------------------------63
4. Conclusions------------------------------------------------------------------------------------64
Chapter 5: Fabrication of Ag@SiO2@Agseed with Ag@SiO2@AgNPs Core-Shell Nanohybrids
for Photothermal therapy-PTT and Bioimaging of Cancer cells----------------------------------65
1. Introduction------------------------------------------------------------------------------------66
2. Experimental Section ------------------------------------------------------------------------69
2.1. Materials-------------------------------------------------------------------------69
2.2. Synthesis of silver nanoparticles ---------------------------------------------69
2.3. Synthesis of Ag@SiO2@Agseed nanoparticles ------------------------------70
2.4. Synthesis of Ag@SiO2@AgNPs nanoparticles ------------------------------70
2.5. Characterization ----------------------------------------------------------------70
2.6. Fluorescence image of Ag@SiO2@Agseed, Ag@SiO2@AgNPs core-shell
nanoparticles -------------------------------------------------------------------101
2.7. Cell viability assay -------------------------------------------------------------71
2.8. Photothermal efficiency of Ag@SiO2@Agseed, Ag@SiO2@AgNPs coreshell nanoparticles--------------------------------------------------------------72
xiii
2.9. Cell apoptosis imaging---------------------------------------------------------72
3. Results and discussion -----------------------------------------------------------------------73
3.1. Characterization of Ag@SiO2@Agseed, Ag@SiO2@AgNPs Core-Shell
Nanoparticle ---------------------------------------------------------------------73
3.2. Morphology of Ag@SiO2@Agseed, Ag@SiO2@AgNPs Core-Shell
Nanoparticle---------------------------------------------------------------------76
3.3. Photothermal therapy efficacy of Ag@SiO2@Agseed and
Ag@SiO2@AgNPs nanoparticles----------------------------------------------80
4. Conclusion ------------------------------------------------------------------------------------88
Chapter 6: Summary and Conclusions ---------------------------------------------------------------89
List of Publications------------------------------------------------------------------------------------------91
Conference Presentation------------------------------------------------------------------------------------92
References ----------------------------------------------------------------------------------------------------93
                                

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