甲基丙烯醯氧乙基三甲基氯化銨(TMA)和3-磺酸丙基甲基丙烯酸鉀鹽(SA)之混合電荷類雙離子sulfobeaine共聚高分子表面抗蛋白質吸附之研究

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許世煌(Shih-huang Hsu) 查詢紙本館藏

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化學工程與材料工程學系

論文名稱

甲基丙烯醯氧乙基三甲基氯化銨(TMA)和3-磺酸丙基甲基丙烯酸鉀鹽(SA)之混合電荷類雙離子sulfobeaine共聚高分子表面抗蛋白質吸附之研究
(Studies of Proteins Adsorption Resistance of [2-(Methacryloyloxy)ethyl]trimethylammonium(TMA) and 3-Sulfopropyl methacrylate potassium salt(SA) Mixed-charged Copolymer Brushs of Sulfobetaine-like Surface)

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摘要(中)

對於生醫材料控制其表面達到抗非特異性生物分子吸附的特性是重要的一環，並且材料表面除了表現出優異的抗吸附能力還必須擁有長效的穩定性以提供更進一步的實際應用。
本研究選用[2-(Methacryloyloxy)ethyl]trimethylammonium (TMA) 和3-Sulfopropyl methacrylate potassium salt (SA)單體，在修飾有溴官能基的金膜表面聚合成混合電荷共聚刷狀高分子poly (TMA/SA)。在單體比例1:1總濃度1.2M室溫的高分子聚合條件下可以得到表面為電中性的混合電荷共聚刷狀高分子；三種蛋白質Human Serum Albumin (HSA)、γ-Globulin及Fibrinogen於不同表面的非特定吸附行為由表面電漿共振儀(surface plasma resonance, SPR)來檢測獲得。此外在本研究中三種完美覆蓋的表面CH3-SAM、OEG-SAM以及poly(SBMA)刷狀高分子表面其蛋白質吸附的行為也作為實驗數據上的比較對象。
在探討操作環境對poly (TMA/SA)抗蛋白質吸附的影響方面，本研究使用的環境變因包括：溫度、鹽濃度、鹽種類以及pH值。也發現在這些實驗操作條件下poly(TMA/SA)對三種指標性蛋白質展現良好的穩定性和抗吸附能力，展現了poly(TMA/SA)的使用穩定性。此外本研究除了使用單一蛋白質做吸附測試外，還做了人體血漿及血小板貼附測試，實驗結果顯示poly(TMA/SA)表現出近似poly(SBMA)刷狀高分子良好的抗血漿吸附能力，在血小板貼附的測試中，也未發現活化的血小板貼附於poly(TMA/SA)表面，顯示poly(TMA/SA)具有相當良好的血液相容性。

摘要(英)

An ideal nonbiofouling surface for biomedical applications requires both high-efficient antifouling characteristics in relation to biological components and long-term material stability from biological systems.
This work describes the performance of an antifouling surface with grafted mixed charge copolymer brushes. We form statistical copolymer brush coatings via surface-initiated atom transfer radical polymerization (ATRP) from the bromide-covered gold surface with a comonomer mixture of positively charged [2-(methacryloyloxy) ethyl]trimethylammonium chloride (TMA) and negatively charged 3-sulfopropyl methacrylate potassium salt (SA). The polymerization condition with the molar ratio (TMA:SA) of 1:1 and concentration of 1.2M in the reaction solution at 25oC allowed the formation of a neutral mixed charge polymer brush [poly(TMA/SA)], which was determined by electron spectroscopy for chemical analysis (ESCA).
The nonspecific adsorption of fibrinogen,γ-globulin, and serum albumin from human plama was measured using a surface plasmon resonance (SPR) biosensor. Three controlled surfaces with methyl-terminated (CH3) self-assembled monolayers (SAMs), oligo(ethylene glycol)-terminated (OEG) SAMs and well-packed polyzwitterionic sulfobetaine methacrylate (polySBMA) brushes were also studied for comparison. It was found that excellent stable nonbiofouling surface with grafted poly(TMA/SA) can be performed with a cycling test of the three model protein adsorption and in a wide range of various salt types, buffer composition, solution pH, and temperature. It was further revealed that the poly(TMA/SA) grafted surface effectively reduces the plasma protein adsorption from platelet poor plasma solution to a level comparable to the well-packed polySBMA grafted surface but superior than that of adsorption on a surface terminated with tetra(ethylene glycol). The adhesion and activation of platelet from platelet rich plasma solution were not observed on the poly(TMA/SA) grafted surface.
This work further concludes that a surface with good hemocompatibility can be simply achieved by the well-packed surface grafted neutral mixed charge poly(TMA/SA) brushes.

關鍵字(中)

★ 雙離子性高分子
★ 抗生物分子吸附
★ 原子自由基轉移聚合法
★ 血液相容性
★ 表面電漿共振
★ 蛋白質

關鍵字(英)

★ zwitterionic
★ sulfobetanie
★ SPR
★ protein
★ nonbiofouling
★ ATRP
★ hemocompatibility

論文目次

中文摘要…………………………………………………..……………………………........I
Abstract………………………………………….………………………………………….Ⅱ
誌謝…………………………………………..…………………………………………….Ⅳ
目錄………………………………………………………………………………….……...V
圖目錄…………………………………………………………………………… .……..VIII
表目錄……………………………………………………………..………...………….....X I
第一章緒論…………………………………………………………..…………………1
第二章文獻回顧……………………………………………………………………..…3
2.1 表面電漿共振 (Surface Plasmon Resonance,SPR) …………………….…….…...3
2.1.1 表面電漿共振原理…………………………………...…..………………..3
2.1.2 表面電漿共振的應用……………………….………………..……………7
2.1.3 表面電漿共振感測儀類型………………..……………………………….7
2.1.4 表面電漿共振感測儀檢測生物反應………………………...……………8
2.2 原子轉移自由基聚合法……………………………………..…………...……….10
(Atom Transfer Radical Polymerization, ATRP)
2.3 生物相容性材料………………………………………………………. ...……….13
2.3.1 生物相容性………………………………………..………………...……14
2.3.2 組織相容性………………..……..…...…………………………..………16
2.3.2.1生物材料與發炎………..……..……………..………………...…17
2.3.2.2生物材料與腫瘤………..……..……………..………………...…17
2.3.3血液相容性………………..……..……………………………..…………18
2.3.3.1血液之組成……………..……..……………..……………...……21
2.3.3.2血液組成與表面的交互作用.………………..…………………..23
2.3.3.3材料與血液的相互關係…….………………..…………………..26
2.3.3.4血小板……………………….………………..………………….27
2.3.3.5生物材料與血小板的相互關係…….…..……………………….29
2.3.3.6凝血機制……...…..……………………………………………...29
2.3.3.7生物材料與凝血機制的相互關係………...…………………….33
2.3.4 血液相容性材料之回顧…………………………………………………34
2.3.4.1水凝膠……………………………………………………………35
2.3.4.2氟化表面…………………………………………………………35
2.3.4.3白蛋白之表面塗佈(Albumin Coating) …..……………………...36
2.3.4.4 表面之聚乙烯乙二醇固定化………………..………………….36
2.3.4.5 類磷脂質之仿生表面………………………..….........................37
2.3.4.6 表面之幾丁聚糖固定化……………………...…........................38
2.3.4.7 表面之肝素固定化…………………………...…........................39
2.3.5 仿生雙離子性高分子之血液相容性相關研究…….…...........................41
2.3.5.1 PC類雙離子性高分子………………….……………………….43
2.3.5.2 其他人工合成的雙離子性高分子……….……………………..51
第三章實驗藥品、設備及實驗方法…………………………….…………………..63
3.1 實驗藥品………………………………………………………………....……...63
3.2 實驗設備…………………………………………………....…………………...64
3.3 實驗方法………………………………………………………………………...66
3.3.1 緩衝溶液的製備…………………………………………………………66
3.3.2 蛋白質溶液的製備………………………………………………………67
3.3.3 血漿溶液的分離與稀釋…………………………………………………67
3.3.4 金表面改質………………………………………………………………68
3.3.5 表面電漿共振感測儀之實驗……………………………………………75
3.3.6 血小貼附實驗……………………………………………………………76
第四章結果與討論………………………………………………….. ………………77
4.1 表面鑑定………………………………………………………………………...78
4.1.1 ESCA檢測樣品表面組成和靜態接觸角測量………. …………………78
4.2 蛋白質吸附測試……………………………………………... ………………...80
4.2.1抗單一蛋白質吸附測試………………………………. ………………...81
4.2.2 循環使用穩定性測試……..……………………………………………..85
4.2.3各種效應對poly(SBMA)穩定性的影響……..………. ………………...86
4.2.3.1 鹽類效應………………………………………. ………………...86
4.2.3.2 pH值效應……………………………………. ………………...94
4.3 血漿吸附測試………………………………………………... ………………...97
4.4 血小板貼附測試……………………………………………... ………………...99
4.5長時間保存對刷狀高分子抗蛋白吸附的影響……………... ………………..101
第五章結論………………………………………………………………………….103
第六章參考文獻…………………………………………………………………….105

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指導教授

陳文逸(Wen-Yih Chen)

審核日期

2009-7-29

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