生物啟發兩性雙離子自組裝抗汙塗層｜國立中央大學博碩士論文系統

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研究生：	朱思澔 Sz-Hau Chu
論文名稱：	生物啟發兩性雙離子自組裝抗汙塗層 Bio-inspired Zwitterionic Assemblies for Robust Antifouling Coatings
指導教授：	黃俊仁 Chun-Jen Huang
口試委員:
學位類別：	碩士 Master
系所名稱：	生醫理工學院 - 生物醫學工程研究所 Graduate Institute of Biomedical Engineering
論文出版年：	2015
畢業學年度：	103
語文別：	中文
論文頁數：	94
中文關鍵詞：	非特異性吸附、自組裝薄膜、兩性雙離子氨基酸、光趨動氧化奈米材料、光熱治療
外文關鍵詞：	zwitterionic amino acid, nanomaterials colloidal stability, photo induced oxidation
相關次數：	點閱：16 下載：0
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植入性醫療器材，由於長時間與血液、體液接觸，容易因為表面的蛋白質、微生物的非特異性吸附，進而引發儀器被組織包覆，效能、靈敏性受損，甚至造成凝血反應、微生物感染、血栓形成等問題。而目前的解決此問題的其中一項辦法為在醫療器材表面修飾一層抗貼附塗層以降低非特異性吸附，避免併發症的發生。其中以自組裝 (Self-Assembling)的修飾方法由於其步驟簡單、處理快速等特性尤為受到矚目。然而，也有許多學者認為，自組裝膜的表面穩定性與抗汙功效，會是其使用上的一大問題。
在本研究中，我們利用天然兩性雙離子氨基酸─半胱胺酸(Cysteine)、自行合成之半胱胺酸四級銨衍生物─半胱胺酸甜菜鹼(Cysteine betaine)和市面上常使用之親水性抗貼附材料─聚乙二醇(Polyethylene Glycol)，自組裝於金基板上，利用水接觸角、X射線光電子能譜、掃描式隧道顯微鏡觀察修飾後表面的親水性質、表面元素組成及分子排列情形。並藉由細胞毒性試驗MTT、革蘭氏陽性菌表皮葡萄球菌（Staphylococcus epidermidis）和陰性菌綠膿桿菌（Pseudomonas aeruginosa）細菌貼附、3T3纖維母細胞貼附和蛋白質貼附，比較其化學穩定度、應用範圍及抗貼附能力。最後，將其修飾在空心金銀奈米殼(Hollow Ag@Au nanoshells) 上，調控溫度、鹽濃度、離子濃度、蛋白質貼附等變因，觀察其膠體穩定性並應用於近紅外光光熱治療。最後開發出一極具潛力之生物衍生兩性雙離子自組裝抗汙塗層。

Implantable medical devices are widely used in clinical medicine. However, they directly contact to blood or body fluid and nonspecific adsorption of protein or microorganisms may occur. This process will lead to decreased efficiency or sensitivity which further facilitates coagulation, infection or thrombus formation. Modification the surface of medical device with antifouling coatings may decrease the possibility of nonspecific adsorption and prevent formation of complications. Among them, modification by self-assembling is an easy and efficient way to improve the biocompatibility. Nevertheless, the stability and effectiveness of self-assembled monolayers (SAMs) is also an issue for long-term biomedical applications.
In this study, we used cysteine (natural sulfur-containing zwitterionic compounds), cysteine betaine (derivative of cycteine with quaternary ammonium group at its terminal end), and polyethylene glycol (a widely used hydrophilic antifouling material) to decorate with Au substrate through Au-thiol interaction. We arranged contact angle goniometer, X-ray photoelectron spectroscopy (XPS), and scanning tunneling microscope (STM) to examine the hydrophilic property, surface elemental composition and molecular arrangement situation in this study. MTT cytotoxicity assays, bacterial adsorption test, 3T3 fibroblast cel�#(~sorption, and protein adsorption were also arranged to compare the range of application and antifouling capacity. Finally, the modification was applied to hollow gold silver nanoshells. After regulating temperature, ion concentration, and protein adsorption, we tested the colloid stability. This technique then applied to near-infrared photoelectron thermal therapy and developed into bio-derived zwitterionic assemblies for antifouling coatings with high potential in clinical implications.

目錄
中文摘要.................................................. v
Abstract...................................................vi
致謝 ......................................................viii
目錄 ......................................................ix
圖目錄 ....................................................xiii
表目錄 ....................................................xv
簡寫對照表 ................................................xvi
第一章 文獻回顧 ...........................................1
1-1 醫療器械、植入物的生物汙染 ............................1
1-1-1 異物反應(Foreign Body Response)......................1
1-1-2 微生物貼附...........................................2
1-2 抗生物沾黏材料 ........................................3
1-2-2 聚乙二醇材料 ........................................4
1-2-3 雙離子材料 ..........................................7
1-3 自組裝單分子膜 ........................................9
1-3-1 硫醇在金表面之自組裝單分子膜.........................10
1-3-2帶有硫醇分子的天然氨基酸兩性雙離子....................11
1-4 半胱胺酸應用於金表面修飾.............................. 13
1-5 半胱胺酸修飾於金表面所衍生之問題.......................15
1-5-1羧基和氨基尾端的硫醇分子自組裝改進方法 ...............16
1-5-2氨基於金表面上的光氧化情形............................20
1-6 奈米醫學 ..............................................22
1-6-1 金銀奈米殼(Hollow Gold Silver Nanoshells)............23
1-6-2 近紅外光光熱治療.....................................24
第二章 研究目的.... .......................................25
第三章 材料與方法 ...... ..................................26
3-1 實驗藥品...............................................26
3-2 實驗儀器...............................................27
3-3 材料合成 ..............................................28
3-3-1 胱胺酸的四級銨化 ....................................28
3-3-2 半胱胺酸甜菜鹼.......................................28
3-4 材料鑑定及表面分析.....................................29
3-4-1 半胱氨酸和半胱氨酸甜菜鹼的酸鹼滴定...................29
3-4-2 半胱氨酸、半胱氨酸甜菜鹼的毒性試驗 (MTT).............29
3-4-3 兩性雙離子半胱氨酸、半胱氨酸甜菜鹼、聚乙二醇的自組裝膜製備.........................................................30
3-4-4 兩性雙離子半胱胺酸甜菜鹼掃描式穿隧電子顯微鏡.........30
3-4-5 光氧化試驗...........................................31
3-4-6 水接觸角測量.........................................31
3-4-7 高解析電子能譜儀(XPS) 分析...........................31
3-4-8 細菌貼附試驗.........................................32
3-4-9 蛋白質貼附試驗.......................................32
3-4-10 細胞貼附試驗........................................33
3-5 金銀奈米殼之分析及應用.................................34
3-5-1 金銀奈米殼的表面修飾.................................34
3-5-2 光氧化後蛋白質貼附粒徑分析 ..........................35
3-5-3 長時間蛋白質貼附粒徑分析 ............................35
3-5-4 不同pH值蛋白質貼附粒徑分析 ..........................35
3-5-5 銅離子對於UV-VIS吸收光譜之影響.......................36
3-5-6 不同溫度、同一鹽濃度下對於UV-VIS吸收光譜之影響.......36
3-5-7 近紅外光雷射升溫試驗 ................................36
3-5-8 長時間近紅外光雷射升溫試驗...........................37
第四章 實驗結果 ...........................................38
4-1 兩性雙離子半胱氨酸及半胱氨酸甜菜鹼 ....................38
4-1-1 半胱胺酸甜菜鹼NMR頻譜分析 ...........................38
4-1-2 半胱胺酸甜菜鹼 ESI-MS高效能質譜儀分析 ...............40
4-1-3 酸鹼滴定.............................................41
4-1-4 半胱胺酸甜菜鹼STM表面形態分析........................42
4-1-5 MTT細胞毒性試驗......................................43
4-1-6 水接觸角.............................................43
4-1-7 XPS表面元素分析......................................44
4-1-8 光氧化後水接觸角.....................................46
4-1-9 光氧化後XPS 表面元素分析.............................48
4-1-10 半胱氨酸和半胱胺酸甜菜鹼的抗貼附性能................49
4-1-11 兩性雙離子於金銀奈米殼上的膠體穩定性................54
4-2 半胱胺酸甜菜鹼和硫醇聚乙二醇...........................59
4-2-1 水接觸角.............................................59
4-2-2 XPS 表面元素分析.....................................60
4-2-3 半胱胺酸甜菜鹼和硫醇聚乙二醇的抗貼附性能.............61
4-2-4 半胱胺酸甜菜鹼和硫醇聚乙二醇修飾於金銀奈米殼上的膠體穩定性.........................................................63
第五章 討論................................................67
5-1 半胱氨酸的光氧化效應及抗生物沾黏能力的喪失.............67
5-2 聚乙二醇於高鹽濃度、高溫之低耐受性.....................67
5-3 半胱胺酸甜菜鹼的優勢...................................68
5-3-1 pH值的高耐受性.......................................68
5-3-2 胺基光氧化效應的抑制.................................68
5-3-3 半胱胺酸甜菜鹼的抗生物沾黏能力.......................69
5-4 半胱氨酸、聚乙二醇、半胱胺酸甜菜鹼應用於光熱治療.......70
第六章 結論................................................72
第七章 未來展望............................................73
第八章 參考文獻 ...........................................74
                                

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