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研究生：	李友吉 Li, Yu-Chi
論文名稱：	利用微弧陽極氧化法製備具生物活性多孔含鍶鍍層於鉭基材之研究 Porous Coatings Incorporating Strontium on Biomedical Tantalum Through Micro-Arc Oxidation
指導教授：	王清正 Wang, Ching-Cheng
共同指導教授:	李澤民 Lee, Tzer-Min
學位類別：	碩士 Master
系所名稱：	電機資訊學院 - 製造資訊與系統研究所 Institute of Manufacturing Information and Systems
論文出版年：	2014
畢業學年度：	102
語文別：	中文
論文頁數：	106
中文關鍵詞：	微弧陽極氧化、鉭、Ta2O5 、鍶
外文關鍵詞：	Micro-arc oxidation, Tantalum, Ta2O5, Strontium
相關次數：	點閱：84 下載：0
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本研究係利用微弧氧化 (Micro-arc oxidation, MAO)法，利用不同鍶濃度之電解液通入高電壓，於材料表面形成微弧火花進而形成氧化層，將生醫用鉭基材 (ASTM F-560)上披覆上一層含有鍶鈣磷成分之氧化鉭鍍層，並在表面上產生分布均勻的孔洞。目的是為了探討材料表面不同鍶濃度對MC3T3-E1細胞活性之影響。X-ray繞射儀與EDS成分分析儀鑑定披覆鍍層之相組成及成分，LV-SEM觀察經MAO處理後的表面形態與厚度。結果發現，鍍層主要成分為Ta2O5，且在氧化層當中含有鈣磷鍶三種元素。當電壓為200 V以下時，其表面為陽極氧化反應，無微弧氧化之火花產生；而在電壓200 V以上時，表面開始產生微弧氧化之火花現象，且表面反應隨著電壓之提升而增大。經由不同鍶濃度之電解液反應過後之試片表面，所形成之厚度並無明顯差異，平均落在5.03~5.39 μm之間。親水性方面，經MAO處理後的試片，與未處理之材料相比皆有非常良好的親水性。細胞相容性方面，將MC3T3-E1細胞分別培養於基材上，探討其生物活性反應。利用細胞增生測試 (Methylthiazoletetrazolium assay, MTT assay)，將細胞培養於試片1、3與7天，探討其增生能力，結果顯示細胞在鉭基材上與鈦基材相比，有較好之增生能力，生長最為迅速。另外測試細胞分生能力 (Alkaline phosphate activity assay, ALP assay)，當改質過後之鉭片其鍶濃度為5 %時，對較其他試片有最好的分生能力。LV-SEM觀察細胞培養1、3與24小時的細胞形態，發現細胞在改質過後的試片貼附能力最好，表示經由微弧氧化改質過後之試片表面結構有較好的細胞親和性。上述結果顯示鍶元素濃度的不同的確會影響細胞的形態與反應，表面鍶濃度含量低的試片有較好的細胞反應。

Containing Sr-Ca-P (Strontium Calcium phosphorus) ceramics layer were bioactive materials. We used micro-arc oxidation (MAO) to manufacture Ta2O5 with Sr-Ca-P porous coatings on tantalum substrates. The aim of this study was to fabricate porous Sr-Ca-P-containing coatings on the tantalum surface using the MAO technique and evaluate the coatings topography, phase, chemical composition and cell response. We used different instruments to investigate the coatings, including X-ray diffraction, FE-SEM and EDS. The micro-arc oxidation were not begin to reaction at 200 V and begin to reaction at 300 V. The results showed that the thickness of Ta2O5 films was the same between different concentration of strontium. The Sr-Ca-P films were deposited about 5.03~5.39 μm. By contact angle analysis, all specimens showed hydrophilic property. In addition, coatings were evaluated with MC3T3-E1 which incubated on samples. Results in the cell proliferation, the pure tantalum exhibited the best biocompatibility. Sample Ta-5Sr exhibited more ALP activity than other samples. In the cell morphology, MC3T3-E1 had the faster rate accreted on sample of MAO modification. In conclusion, it was showed that the strontium of low concentration affected cell behaviors.

中文摘要.....I
Abstract.....II
Extended Abstract.....III
誌謝.....VII
目錄.....VIII
表目錄.....XI
圖目錄.....XII

第一章 緒論.....1
1.1 研究背景.....1
1.2 研究目的.....3
第二章 文獻回顧.....4
2.1 生醫材料簡介.....4
2.1.1 生醫材料定義.....5
2.1.2 生醫材料特性.....5
2.1.3 生醫材料分類.....6
2.1.4 牙科生醫材料之需求.....8
2.2 鉭金屬於生醫應用簡介.....9
2.3 鍶元素介紹.....10
2.4 披覆生物活性陶瓷製程技術.....11
2.4.1 電漿熔射法.....12
2.4.2 濺鍍披覆法.....12
2.4.3 溶膠-凝膠法.....13
2.4.4 仿生浸泡法.....13
2.4.5 電化學沉積法.....13
2.4.6 陽極化處理法.....14
第三章 材料與實驗方法.....16
3.1 研究方法.....16
3.1.1 實驗流程.....16
3.1.2 實驗材料.....16
3.2 試片前處理.....19
3.3 微弧氧化陽極處理裝置.....19
3.4 基本性質測試.....20
3.4.1 SEM、EDS表面分析.....20
3.4.2 XRD相分析.....20
3.4.3 ICP-OES分析.....21
3.4.4 橫截面分析.....21
3.4.5 微硬度測試.....21
3.4.6 粗糙度測試.....22
3.4.7 表面孔徑分析.....22
3.4.8 親疏水性測試.....23
3.4.9 活性測試.....23
3.5 體外實驗.....24
3.5.1 滅菌處理.....24
3.5.2 細胞培養.....24
3.5.3 細胞增生活性測試.....25
3.5.4 細胞分化活性測試.....26
3.5.5 細胞礦化活性測試.....27
3.5.6 細胞表面形態觀察.....28
3.5.7 細胞免疫螢光染色.....29
3.5.8 統計分析.....31
第四章 結果與討論.....32
4.1 基本性質探討.....32
4.1.1 SEM/EDS表面分析結果.....32
4.1.2 XRD相分析結果.....34
4.1.3 ICP-OES元素分析.....35
4.1.4 橫截面分析結果.....36
4.1.5 微硬度測試結果.....36
4.1.6 粗糙度測試結果.....37
4.1.7 表面孔徑觀察結果.....37
4.1.8 親疏水性測試結果.....38
4.1.9 活性測試結果.....38
4.2體外實驗.....40
4.2.1 細胞增生活性分析結果.....41
4.2.2 細胞分化活性分析結果.....41
4.2.3 細胞礦化活性分析結果.....42
4.2.4 細胞表面形態觀察結果.....42
4.2.4 細胞免疫螢光染色結果.....44
第五章 結論.....45
參考文獻.....46
 
表目錄
表2.1	鉭金屬之物理性質.....56
表3.1	ASTM F-560醫療級純鉭成分表.....57
表3.2	人工模擬體液 (SBF)之成分 (ISO 23317).....57
表4.1	各種不同Sr濃度之電解液配方.....58
表4.2	各種不同Sr濃度試片EDS面分析.....59
表4.3	ICP-OES之定性分析結果.....60
表4.4	0 % Sr試片浸泡人工模擬體液後試片Ca及P成分EDS面分析.....61
表4.5	1 % Sr試片浸泡人工模擬體液後試片Ca及P成分EDS面分析.....61
表4.6	5 % Sr試片浸泡人工模擬體液後試片Ca及P成分EDS面分析.....62
表4.7	10 % Sr試片浸泡人工模擬體液後試片Ca及P成分EDS面分析.....62
表4.8	25 % Sr試片浸泡人工模擬體液後試片Ca及P成分EDS面分析.....63
表4.9	細胞增生 (MTT)一天之SPSS統計分析表.....64
表4.10	細胞增生 (MTT)三天之SPSS統計分析表.....65
表4.11	細胞增生 (MTT)七天之SPSS統計分析表.....66
表4.12	細胞分化 (ALP)十天之SPSS統計分析表.....67
 
圖目錄
圖1.1	生醫材料於人體之應用.....68
圖2.1	金屬元素的生物相容性.....69
圖2.2	純金屬、不銹鋼及Co-Cr合金之生物相容性與耐蝕性的比較.....70
圖2.3	鍶元素於細胞內之機制.....71
圖2.4	電漿噴射示意圖.....71
圖2.5	電解沉積示意圖.....72
圖2.6	表面均勻且有顏色變化之微弧氧化表面.....73
圖2.7	MC3T3-E1細胞貼附(a)未改質之鈦基材 (b)微弧氧化後之鈦基材.....73
圖2.8	鉭金屬微弧陽極氧化即時圖 (a) 10 s; (b) 20 s; (c) 1 min; (d) 3 min; (e) 5 min; (f) 15 min; (g) 30 min; (h) 45 min.....74
圖2.9	微弧氧化後鍍層與基材之橫截面.....75
圖3.1	實驗流程圖.....76
圖3.2	處理裝置示意圖.....77
圖3.3	試片硬度值取樣示意圖.....77
圖4.1	微弧陽極氧化設備實體圖.....78
圖4.2	微弧陽極氧化電流與時間關係圖.....79
圖4.3	磷酸二氫鈉、醋酸鈣電解液之微弧陽極氧化一分鐘後巨觀圖 (a)100 V；(b)200 V；(c)300 V；(d)400 V；(e)500 V.....80
圖4.4	磷酸二氫鈉、醋酸鈣電解液之微弧陽極氧化後SEM圖:(A,a)100 V/1 min；(B,b)200 V/1 min；(C,c)300 V/1 min；(D,d)400 V/1 min；(E,e)500 V/1 min；(A-E x500；a-e 2000x).....80
圖4.5	磷酸二氫鈉、醋酸鈣及醋酸鍶電解液之微弧陽極氧化後巨觀圖(a) 0 % Sr；(b) 1 % Sr；(c) 5 % Sr；(d) 10 % Sr；(e) 25 % Sr.....81
圖4.6	磷酸二氫鈉、醋酸鈣及醋酸鍶電解液之微弧陽極氧化後SEM圖: (A,a) 0 % Sr；(B,b) 1 % Sr；(C,c) 5 % Sr；(D,d) 10 % Sr；(E,e) 25 % Sr；(A-E x500；a-e 2000x).....82
圖4.7	各種不同Sr濃度試片表面EDS面分析: (a) 0 % Sr；(b) 1 % Sr；(c) 5 % Sr；(d) 10 % Sr；(e) 25 % Sr.....83
圖4.8	各種不同Sr濃度試片之取代率 (at %).....83
圖4.9	純鉭及磷酸二氫鈉、醋酸鈣電解液之微弧陽極氧化後TF-XRD圖.....84
圖4.10	純鉭及各種不同Sr濃度電解液之微弧陽極氧化後TF-XRD圖.....84
圖4.11	各種不同Sr濃度試片之橫截面SEM圖: (a) 0 % Sr；(b) 1 % Sr；(c) 5 % Sr；(d) 10 % Sr；(e) 25 % Sr (5000x).....85
圖4.12	各種不同Sr濃度試片表面之微硬度.....86
圖4.13	各種不同Sr濃度試片表面之粗糙度.....87
圖4.14	各種不同Sr濃度試片表面3D圖:(a) Ti；(b) Ta；(c) 0 % Sr；(d) 1 % Sr；(e) 5 % Sr；(f) 10 % Sr；(g) 25 % Sr (50x).....88
圖4.15	微弧陽極氧化後表面孔徑分析統計柱狀圖.....89
圖4.16	液滴與試片親疏水性測試巨觀圖:(a) Ti；(b) Ta；(c) 0 % Sr；(d) 1 % Sr；(e) 5 % Sr；(f) 10 % Sr；(g) 25 % Sr (50x).....90
圖4.17	各種不同Sr濃度試片表面之接觸角.....91
圖4.18	不同Sr濃度試片浸泡SBF 0天之SEM圖: (A,a) Ti；(B,b) Ta；(C,c) 0 % Sr；(D,d) 1 % Sr；(E,e) 5 % Sr；(F,f) 10 % Sr；(G,g) 25 % Sr；(A-G 200x；a-g 1000x).....92
圖4.19	不同Sr濃度試片浸泡SBF 1天之SEM圖: (A,a) Ti；(B,b) Ta；(C,c) 0 % Sr；(D,d) 1 % Sr；(E,e) 5 % Sr；(F,f) 10 % Sr；(G,g) 25 % Sr；(A-G 200x；a-g 1000x).....93
圖4.20	不同Sr濃度試片浸泡SBF 3天之SEM圖: (A,a) Ti；(B,b) Ta；(C,c) 0 % Sr；(D,d) 1 % Sr；(E,e) 5 % Sr；(F,f) 10 % Sr；(G,g) 25 % Sr；(A-G 200x；a-g 1000x).....94
圖4.21	不同Sr濃度試片浸泡SBF 7天之SEM圖: (A,a) Ti；(B,b) Ta；(C,c) 0 % Sr；(D,d) 1 % Sr；(E,e) 5 % Sr；(F,f) 10 % Sr；(G,g) 25 % Sr；(A-G 200x；a-g 1000x).....95
圖4.22	不同Sr濃度試片浸泡SBF 21天之SEM圖: (A,a) Ti；(B,b) Ta；(C,c) 0 % Sr；(D,d) 1 % Sr；(E,e) 5 % Sr；(F,f) 10 % Sr；(G,g) 25 % Sr；(A-G 200x；a-g 1000x).....96
圖4.23	不同Sr濃度試片浸泡SBF 0、1、3及7天後之離子釋放圖.....97
圖4.24	浸泡人工模擬體液7天後之試片TF-XRD圖.....98
圖4.25	Sr濃度5 %試片浸泡SBF 0、1、3、7及21天之TF-XRD圖.....98
圖4.26	MC3T3-E1細胞培養於不同Sr濃度試片1、3及7天之細胞增生圖.....99
圖4.27	MC3T3-E1細胞培養於不同Sr濃度試片10天之細胞分化圖.....100
圖4.28	MC3T3-E1細胞培養於不同Sr濃度試片21天之細胞礦化圖: (A,a) Ti；(B,b) Ta；(C,c) 0 % Sr；(D,d) 1 % Sr；(E,e) 5 % Sr；(F,f) 10 % Sr；(G,g) 25 % Sr；(A-G 沒細胞40x；a-g 有細胞40x).....101
圖4.29	MC3T3-E1細胞培養於不同Sr濃度試片1小時之細胞型態圖: (a) Ti；(b) Ta；(c) 0 % Sr；(d) 1 % Sr；(e) 5 % Sr；(f) 10 % Sr；(g) 25 % Sr (2000x).....102
圖4.30	MC3T3-E1細胞培養於不同Sr濃度試片3小時之細胞型態圖: (a) Ti；(b) Ta；(c) 0 % Sr；(d) 1 % Sr；(e) 5 % Sr；(f) 10 % Sr；(g) 25 % Sr (2000x).....103
圖4.31	MC3T3-E1細胞培養於不同Sr濃度試片24小時之細胞型態圖: (a) Ti；(b) Ta；(c) 0 % Sr；(d) 1 % Sr；(e) 5 % Sr；(f) 10 % Sr；(g) 25 % Sr (2000x).....104
圖4.32	細胞生長型態示意圖: (a) 細胞呈現圓球狀並且沒有伸出觸角；(b)與(c) 細胞開始往側向延伸並且逐漸攤開；(d) 細胞完全攤開並且開始往周圍生長.....105
圖4.33	MC3T3-E1細胞培養於不同Sr濃度試片3小時之免疫螢光圖: (a) Ti；(b) Ta；(c) 0 % Sr；(d) 1 % Sr；(e) 5 % Sr；(f) 10 % Sr；(g) 25 % Sr (40x).....106
圖4.34	MC3T3-E1細胞培養於不同Sr濃度試片24小時之免疫螢光圖: (a) Ti；(b) Ta；(c) 0 % Sr；(d) 1 % Sr；(e) 5 % Sr；(f) 10 % Sr；(g) 25 % Sr (40x).....107
                                    

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