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研究生:葉錫誼
研究生(外文):Hsi-Yi Yeh
論文名稱:NiTi記憶金屬表面之血栓調節素固定化及其生物活性與血液相容性之研究
論文名稱(外文):Bioactivity and Platelet Adhesion Studies for Thrombomodulin-immobilized Nitinol Surface
指導教授:林睿哲
指導教授(外文):Jui-Che Lin
學位類別:碩士
校院名稱:國立成功大學
系所名稱:化學工程學系碩博士班
學門:工程學門
學類:化學工程學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:77
中文關鍵詞:NiTi合金血栓調節素表面改質血液相容性血小板吸附
外文關鍵詞:nitinolthrombomodulinblood compatibilitysurface
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NiTi合金是一種新的生醫材料,具有形狀記憶效應,可藉著溫度改變造成的相變化來記憶不同的形狀,使其在外科手術上具有便利性,此外更具有良好的生物相容性,在植入人體後,不易引起生理的免疫反應,因此正逐漸被廣泛的應用在臨床醫療上。然而其本身有一顯著的缺點,即對血液的相容性不佳,容易導致血栓的發生。
在本實驗中,嘗試利用矽烷化反應來對NiTi合金的表面進行改質,使其具有一活性的-NH2官能基,接著利用偶合劑將具有抗凝血功能的血栓調節素,以化學鍵結的方式固定在合金表面上。並利用BCA、Lowry和Brafdord試劑來定量固定化的蛋白質,再藉由血小板吸附實驗及protein C活化測試,評估其生物活性和血液相容性。
實驗結果顯示,固定化後的血栓調節素仍然具有促使凝血酶活化protein C的能力,但與溶液相中的血栓調節素相比,固定化後的血栓調節素有活性降低的現象,經由血小板吸附實驗,可以發現表面固定血栓調節素後的NiTi合金,明顯減少了對血小板的吸附情形,因此可以證明經由血栓調節素的固定化後,確實能夠改善NiTi合金的血液相容性。
Nitinol is a newly developed biomaterial and is gaining popularity in many biomedical applications. Because nitiol has good biocompatibility, it will not induce the inflammatory response and the repulsion by the immunization after implanted in human body. Besides, nitinol is a kind of shape memory alloy that can memorize shapes under different temperature. This can improve the convenience in surgery. However, nitinol has poor blood compatibility that limits the applications of nitinol.

In this study, the surface of nitinol was modified by silanization with organosilane to introduce amino groups. The introduced functional groups were available for the subsequent covalent immobilization of thrombomodulin (TM) by using coupling reagent. The immobilized TM were determined by BCA method, Lowry method, and Braford method. The blood compatibility of various sample and the bioactivity of the immobilized TM were evaluated by protein C assay and platelet adhesion test.

Based on the result, it was found that the immobilized TM still had the ability to accelerate the activation of protein C, while its activity was lower than the free TM in solution. Furthermore, platelet adhesion test showed that the TM-immobilized surface had the least platelet adhesion. Therefore, it is possible to improve the blood compatibility of nitinol by the immobilization of thrombomodulin on surface.
目 錄
中文摘要……………………………………………….………...………...Ⅰ
英文摘要……………………………………………………….…...……...Ⅱ
誌謝………………………………………………………………………...Ⅲ
目錄………………………………………………………………….…..…Ⅳ
表目錄……………………………………………………….……………..Ⅶ
圖目錄…………………………………………………..…………….……Ⅷ
主文
第一章 前言 …………………………...……………..……………………1
第二章 凝血機制 ……………………...……………..……………………3
2.1 血液的組成. .…………………...……………..……………………3
2.2 血小板的機能…………………...……………..……………………4
2.3 凝血機制………………………...……………..……………………6
2.3.1內因路徑….……………………………….……………………7
2.3.2 外因路徑…………………...……………..……………………8
2.4 血塊的形成 …..………………...……………..……………………9
2.5 纖維蛋白溶解系統……………...……………..……………………9
第三章 血栓調節素的特性…………...………………..…………………18
3.1 血栓調節素的性質….………...…………………..……………… 18
3.2 血栓調節素的結構 …………...……………..……………………18
3.3 血栓調節素的功能 …………...……………..……………………19
3.3.1 Protein C的活化 ……………………..…………………..19
3.3.2 抑制凝血酶活化其他因子的功能 …….....…………………20
第四章 文獻回顧 …………………………………………...………..…..25
4.1 NiTi合金的組成與性質…………………………………..……… 25
4.2 形狀記憶效應…………………………………………...…………26
4.3 超彈性……………………...……………..………………………..26
4.4 NiTi合金使用上的注意事項………….…………….………….. 27
4.5 NiTi合金在生醫材料上的應用…………..……………………… 27
4.6 對NiTi合金的表面改質 ………………………….……………..29
4.7 研究動機與目的……………………………….….……………… 30
第五章 實驗內容…………………………………….…………...…….…35
5.1實驗藥品 …………………………………….…………………….35
5.2 實驗步驟………………………………………..………………… 39
5.2.1 NiTi合金的前處理………………………..………………….39
5.2.2 表面之矽烷化反應…………………….….……...…………..39
5.2.3 測量表面氨基的數量……………………….…….…...……..40
5.2.4 血栓調節素的固定化…………………....…………………..41
5.2.4.1 以戊二醛為活化劑的固定化方法……....……………..41
5.2.4.2 以diethylene glycol diglycidyl ether
為活化劑的固定化方法……………...………..………..42
5.2.4.3 芳基化反應………………………….…………………..43
5.2.5 固定化蛋白質之定量………………………….……………..44
5.2.5.1 以BCA Method定量…….………………………………44
5.2.5.2 以Lowry Method定量…………….……………………45
5.2.5.3 以Braford Method定量….……………………………46
5.2.6 以Protein C assay 評估血栓調節素的生物活性…….…. 46
5.2.7 血小板吸附性質之測試 …………………………….………47
第六章 結果與討論……………………………………………….………55
6.1 NiTi合金的清洗時間 ………………………..……..……………55
6.2 酸液浸泡時間對矽烷化反應的影響…………..…...……………..56
6.3 接觸角之分析……………..……………………...………………..56
6.4 蛋白質反應濃度與表面固定量的關係…………….....…………..58
6.5 固定化血栓調節素的活性測試……………………….....………..60
6.6 血小板吸附實驗……………………………………..…...………..62
第七章 結論與未來展望………………………………….....……………71
參 考 文 獻 ………………………………………………...……..……..72
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