臺灣博碩士論文加值系統

當鈦植體被植入體內時，血液是第一個接觸到植體材料表面的物質，並且會馬上引發凝血反應來幫助傷口癒合及後續細胞組織的生長，因此血液的凝結是初期骨癒合過程中重要的第一步。目前在生醫工程的領域中對於材料表面誘發凝血作用的機制並不是非常的瞭解。一般認為，金屬材料表面所引發的內在因子凝血路徑的活化，是由於極性材料表面所帶有的負電，誘發凝血第12因子活化而觸發一系列接觸活化作用，最後形成的網狀纖維蛋白與活化血小板構成血塊而達到凝血的作用，但對於鈦植體材料表面的物理及化學性質如何影響凝血反應的機制仍不甚明瞭。
本研究之主要目的即是針對鈦金屬材料經過表面處理後常見的兩種二氧化鈦薄膜晶相結構：銳鈦礦相及金紅石相，將此兩種晶相依不同比例以物理氣相沈積法製備於鈦基材表面，並分別就其表面性質進行表面形貌、薄膜厚度、二氧化鈦晶相結構、以及二氧化鈦薄膜表面自由能與介電常數的分析，接著於不同晶相結構的試片表面進行其全血凝固和第12凝血因子的接觸活化分析、纖維蛋白原、纖維蛋白與血小板的貼附觀察比較，藉由觀察不同二氧化鈦晶相結構薄膜表面性質的改變來比較其對凝血作用的影響，並探討晶相結構對表面電性的影響與凝血作用活化機制的關係。
實驗結果顯示，不同晶相結構之材料表面自由能雖差異不大，但隨著金紅石相含量增加，二氧化鈦薄膜的介電常數會由銳鈦礦相的24.9大幅上升至金紅石相的143.7，表示二氧化鈦薄膜的電容量會隨著銳鈦礦相轉變成金紅石相而提高。而於凝血性質分析的部分則發現，隨著銳鈦礦相至金紅石相的相變化轉移，全血於不同晶相結構材料表面凝集情況的差異雖然不明顯，但二氧化鈦薄膜表面纖維蛋白及血小板的貼附量卻上升，且貼附聚集和細胞攤平及觸角延伸的情況也較明顯，同時也觀察到金紅石相表面所活化第12凝血因子的量也較高。因此根據實驗結果認為，當二氧化鈦薄膜因為相變化由銳鈦礦相轉變為金紅石相，其薄膜介電常數（電容值）會大幅提高，而此表面電性的改變會使得材料表面對於第12凝血因子接觸活化作用以及血小板的貼附增加，進而影響後續凝血作用的發生。

When a titanium (Ti) implant is placed in human body, blood will be the first component to interact with implant surface, which an immediate blood coagulation process will be activated, helping the continued wound healing and tissue cell growth at the implant site, so one could say blood coagulation is an important first step for the early stage of osseointergration. But the mechanism of blood coagulation process activated by material surfaces is not entirely understood nowadays. Generally speaking as the intrinsic coagulation pathway will be activated by negatively charged Ti material surface, which a blood coagulation factor XII will first be contact-activated, then a chain of coagulation factors will be trigger and reacted, ultimately forming a bunch of fibrin mesh and binding with other activated platelets and red blood cells to form blood clots. However, the mechanism of how the physical, chemical and electrical properties of Ti implant surface effect the coagulation reaction still remains unclear.
The aim of this study was to estimate the blood coagulation properties of two kinds of TiO2 crystal structure: anatase and rutile, which happen to have tremendous differences in their surface electrical properties, and by investigating the surface free energy and dielectric constant of both thin film surfaces, we may compare and evaluate the effect of surface electric properties to the hemocompatiblity of TiO2 films.
The results show that, after PVD surface modification, although the surface free energy didn’t show obvious difference between anatase and rutile, but along with increase amount of rutile structure, the dielectric constant increased tremendously, as the activation of FXII, adsorption of fibrinogen, formation of fibrin network and adhesion of platelet, all show the same tendency.

論文電子檔著作權授權書 …………………………………… i
論文審定同意書 ……………………………………………… ii
誌謝 …………………………………………………………… iii
中文摘要 ……………………………………………………… iv
英文摘要 ……………………………………………………… vi
目錄 …………………………………………………………… viii
表目錄 ………………………………………………………… x
圖目錄 ………………………………………………………… xi

第一章 研究背景與目的 …………………………… 1
1.1生醫材料表面特性與生物相容性之關係……………… 1
1.2生醫材料的凝血性質…………………………………… 2
1.3 金屬植體表面之凝血作用…………………………… 3
1.4 鈦金屬之特性………………………………………… 5
1.5 具生物相容性的結晶性二氧化鈦薄膜……………… 6
1.6 研究目的……………………………………………… 7

第二章 研究材料與方法 ………………………… 9
2.1 儀器與試劑…………………………………………… 9
2.2 二氧化鈦薄膜製備…………………………………… 10
2.3 材料表面性質分析…………………………………… 11
2.3.1 表面形貌分析 …………………………………… 11
2.3.2 薄膜厚度分析 …………………………………… 12
2.3.3 晶相結構鑑定 …………………………………… 12
2.3.4 表面自由能量測 ………………………………… 13
2.3.5 介電常數計算 …………………………………… 15
2.4 凝血試驗分析………………………………………… 16
2.4.1 全血凝固試驗 …………………………………… 17
2.4.2 接觸活化第12凝血因子試驗 ………………… 18
2.4.3 纖維蛋白酶原吸附量測 ………………………… 19
2.4.4 纖維蛋白貼附觀察 ……………………………… 19
2.4.5 血小板貼附試驗 ………………………………… 20

第三章 結果 ……………………………………… 23
3.1 二氧化鈦薄膜製備…………………………………… 23
3.2 材料表面性質分析…………………………………… 23
3.2.1 表面形貌分析 …………………………………… 23
3.2.2 薄膜厚度分析 …………………………………… 23
3.2.3 晶相結構鑑定 …………………………………… 24
3.2.4 表面自由能量測 ………………………………… 24
3.2.5 介電常數計算 …………………………………… 25
3.3凝血試驗分析………………………………………… 26
3.3.1 全血凝固試驗 …………………………………… 26
3.3.2 接觸活化第12凝血因子試驗 ………………… 26
3.3.3 纖維蛋白酶原吸附量測 ………………………… 27
3.3.4 纖維蛋白貼附觀察 ……………………………… 28
3.3.5 血小板貼附試驗 ………………………………… 28

第四章 討論 …………………………………… 30
4.1 二氧化鈦之相變化與電性的關係………………… 30
4.2 材料表面電性與凝血作用的關係………………… 32
4.2.1表面電性對蛋白吸附的影響 …………………… 32
4.2.2材料表面蛋白吸附對凝血作用的影響 ………… 34
4.2.3 高介電常數材料表面對第12凝血因子接觸活化作用的影響 36
4.2.4 高介電常數材料表面對血小板貼附的影響 ……… 36

第五章 結論 …………………………………… 38

第六章 未來方向 ……………………………… 39

參考文獻 ……………………………………………… 41
附表 …………………………………………………… 45
附圖 …………………………………………………… 48

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