無鎳鋯基及鈦基金屬玻璃生物相容性之研究

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張順富(Shun-fu Chang) 查詢紙本館藏

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機械工程學系在職專班

論文名稱

無鎳鋯基及鈦基金屬玻璃生物相容性之研究
(Biocompatibility study of Ni-free Zr-base and Ti-base metallic glasses)

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★ 探討不同結晶率對鋯鋁鈷塊狀非晶質合金機械性質之影響	★ 鈦基非晶填料應用於Ti-6Al-4V合金硬焊之微結構及機械性能研究

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摘要(中)

醫療植入物因需長期使用，安全性及可靠性是產品的首要條件。金屬玻璃因其優異的機械性質、耐蝕性、抗菌性與良好的生物相容性，近年來許多學者開始進行將其應用於醫療器械的相關研究。
本研究以真空吸鑄法成功製備出Zr42Cu42Al8Ag8、Zr48Cu36Al8Ag8及Ti40Zr10Cu36Pd14無鎳金屬玻璃，分別利用EDS、XRD、AFM進行成份定量分析、晶體結構以及表面形貌觀察。無鎳金屬玻璃均為非晶結構，其表面粗糙度皆在2.68 nm以下。為評估無鎳金屬玻璃的生物相容性，將小鼠纖維母細胞株L929培養於金屬玻璃上進行觀察細胞貼附於金屬玻璃之型態、細胞毒性測試與金屬離子釋出對細胞影響之分析。其中以Ti40Zr10Cu36Pd14無鎳金屬玻璃生物相容性表現最佳，無論在細胞培養直接接觸法或MTT assay均證實不具有細胞毒性；細胞貼附實驗結果顯示細胞易於生長且細胞膜延展佳，與現行醫療用鈦合金(Ti-6Al-4V)有相似的表現；且其無鎳的成份及極低銅離子溶出比現有醫療用合金具有更良好的生物相容性。

摘要(英)

Safety and reliability are crucial issues to medical instruments and implants. In the past few decays, bulk metallic glass draw lots attentions due to their excellent mechanical properties, good corrosion resistance, antibacterial and good biocompatibility, and were considered as an ideal candidate for medical related implants.
Three Ni-free bulk metallic glasses include the Zr42Cu42Al8Ag8, Zr48Cu36Al8Ag8 and Ti40Zr10Cu36Pd14 were studied and evaluated their potential as medical implants. The glassy nature and composition of these three bulk metallic glasses were firstly confirmed by X-ray diffraction analysis and energy dispersive spectrometer. The surface roughness examined by atomic force microscope, all samples perform nano-meter-scale surface roughness. Several biocompatibility tests were carrying out to evaluate these three bulk metallic glasses co-cultural with L929 murine fibroblast cell line. The results of cellular adhesion behavior, cytotoxic, and metallic ion release affection. The optima biocompatibility results occur at Ti40Zr10Cu36Pd14 bulk metallic glass, cell still attached on the petri dish with good adhesion and exhibit the spindle shape after direct contact test. Furthermore, the Ti40Zr10Cu36Pd14 sample showed the lowest Ni and Cu ion release level which correlate to MTT results. Based on the data mentioned above, we believe that Ti40Zr10Cu36Pd14 bulk metallic glass can be one of ideal candidates for medical implant materials.

關鍵字(中)

★ 無鎳鋯基金屬玻璃
★ 無鎳鈦基金屬玻璃
★ 生物相容性
★ 小鼠纖維母細胞

關鍵字(英)

★ Ni-free Zr-base metallic glass
★ Ni-free Ti-base metallic glass
★ Biocompatibility
★ Murine Fibroblast

論文目次

目錄
摘要 I
Abstract II
致謝 III
目錄 VI
表目錄 IX
圖目錄 X
第一章前言 1
第二章理論基礎 4
2-1 金屬玻璃發展 4
2-2 實驗歸納法 6
2-2-1 合金應包含三種以上的元素 6
2-2-2 主要元素彼此的原子半徑差異須達12 %以上 6
2-2-3 主要成分之混和熱必須為負值 7
2-3 金屬玻璃之製造方法 7
2-3-1 氣態轉固態 7
2-3-2 液態轉固態 8
2-3-3 固態轉固態 8
2-4 金屬玻璃之種類 9
2-5 金屬玻璃之特性 10
2-5-1 機械性質 10
2-5-2 化學性質-耐蝕性 11
2-5-3 磁性質 11
2-5-4 金屬玻璃於醫療器材之應用 12
2-5-5 其他性質 13
2-6 生物相容性 13
2-6-1 細胞毒性評估 14
2-6-2 細胞存活率分析 15
2-6-3 纖維母細胞 15
第三章實驗步驟與方法 17
3-1 實驗流程 17
3-2 合金材料製備 18
3-2-1 合金鑄錠配製 18
3-2-2 真空電弧熔煉 18
3-2-3 真空吸鑄製程 19
3-3 塊狀無鎳金屬玻璃製備 19
3-3-1 慢速切割及線切割製程 19
3-3-2 研磨及拋光製程 19
3-4 微觀組織分析 20
3-4-1 X光繞射分析(XRD) 20
3-4-2 能譜散佈光譜儀分析(EDS) 21
3-4-3 原子力顯微鏡分析(AFM) 21
3-5 細胞培養 21
3-5-1 儀器介紹 22
3-5-2 材料及試劑 23
3-5-3 培養基製備 24
3-5-4 解凍細胞 25
3-5-5 細胞繼代 25
3-5-6 細胞計數 26
3-6 細胞毒性測試 26
3-6-1 固定細胞及臨界點乾燥 26
3-6-2 直接接觸法 28
3-6-3 細胞存活率分析(MTT assay) 29
3-7 金屬離子溶出檢測 31
3-7-1 感應耦合電漿質譜分析法(ICP-MS) 31
第四章結果與討論 33
4-1 成分分析 33
4-2 晶體結構分析 33
4-3 表面粗糙度分析 33
4-4 細胞附著型態的觀察 34
4-5 細胞毒性之直接接觸法分析 34
4-6 細胞毒性之細胞存活率分析(MTT assay) 35
4-7 金屬離子溶出檢測 36
第五章結論 38
參考文獻 40

表目錄
表1-1金屬玻璃之特性[19] 48
表2-1最初金屬玻璃之系統分類[51] 49
表2-2多元系塊狀金屬玻璃種類與發展歷程[63] 50
表2-3多元系塊狀金屬玻璃之成分分類[63] 51
表2-4金屬玻璃之磁特性[56] 52
表3-1細胞資料單[67] 53
表3-2萃取液計算標準表[35] 53
表4-1 Zr42Cu42Al8Ag8、Zr48Cu36Al8Ag8及Ti40Zr10Cu36Pd14無鎳金屬玻璃EDS元素分析 54
表4-2直接接觸法測試定性分析表[35] 55
表4-3 Zr42Cu42Al8Ag8、Zr48Cu36Al8Ag8及Ti40Zr10Cu36Pd14無鎳金屬玻璃直接接觸法定性測定結果 55
表4-4 Zr42Cu42Al8Ag8、Zr48Cu36Al8Ag8及Ti40Zr10Cu36Pd14無鎳金屬玻璃細胞存活率MTT assay測定結果 56
表4-5美國藥典製劑元素雜質規範表[64] 57
表4-6 MTT assay 1天萃取培養基ICP-MS元素分析 58
表4-7 MTT assay 3天萃取培養基ICP-MS元素分析 58
表4-8 MTT assay 5天萃取培養基ICP-MS元素分析 58

圖目錄
圖1-1 結晶材料顯微結構圖[40] 59
圖1-2 金屬玻璃顯微結構圖[40] 59
圖2-1 雙輪連續急冷法[38] 60
圖2-2 合金系統之分類圖[63] 60
圖2-3 結晶與非結晶之X光繞射結果[40] 61
圖2-4 金屬玻璃受外力之分子移動型態[54] 61
圖2-5 金屬玻璃之機械強度比較[55] 62
圖2-6 醫療器材之生物相容性評估架構表[35] 63
圖2-7 細胞代謝產生formazan之反應圖[68] 64
圖3-1 實驗流程圖 65
圖3-2 數位電子天秤 66
圖3-3 電弧熔煉爐 66
圖3-4 氬焊機 67
圖3-5 合金鑄錠外觀：鈦基(左)，鋯基(右) 67
圖3-6 無鎳金屬玻璃板材外觀：鈦基(左)，鋯基(右) 67
圖3-7 電腦數值控制線切割機外觀 68
圖3-8無鎳金屬玻璃塊材拋光後外觀：(左) Zr42Cu42Al8Ag8，(中) Zr48Cu36Al8Ag8，(右)Ti40Zr10Cu36Pd14 68
圖3-9 對照組金屬及合金塊材拋光後外觀：(左) 純鋯，(中) 純銅，(右) Ti-6Al-4V 68
圖3-10 研磨拋光機 69
圖3-11 X光繞射分析儀 69
圖3-12 能量散佈光譜儀 70
圖3-13 原子力顯微鏡 70
圖3-14 無菌操作台 71
圖3-15 光學顯微鏡 71
圖3-16 離心機 72
圖3-17 高溫高壓滅菌釜 72
圖3-18 血球計數盤 73
圖3-19 二氧化碳培養箱 73
圖3-20 低溫震盪培養箱 74
圖3-21 微量盤式分光光譜儀 74
圖3-22 鍍金機 75
圖3-23 臨界點乾燥機 75
圖3-24 掃描式電子顯微鏡 76
圖3-25 感應耦合電漿質譜分析儀 76
圖4-1 Zr42Cu42Al8Ag8、Zr48Cu36Al8Ag8及Ti40Zr10Cu36Pd14無鎳金屬玻璃XRD繞射圖 77
圖4-2 Ti-6Al-4V表面粗糙度(a) 2D (b) 3D 78
圖4-3 Zr42Cu42Al8Ag8無鎳金屬玻璃表面粗糙度(a) 2D (b) 3D 78
圖4-4 Zr48Cu36Al8Ag8無鎳金屬玻璃表面粗糙度(a) 2D (b) 3D 79
圖4-5 Ti40Zr10Cu36Pd14無鎳金屬玻璃表面粗糙度(a) 2D (b) 3D 79
圖4-6 利用SEM以10,000倍率，觀察小鼠纖維母細胞L929貼附各金屬及合金之狀態 80
圖4-7 利用SEM以5,000倍率，觀察小鼠纖維母細胞L929貼附各金屬及合金之狀態 81
圖4-8 利用SEM以3,000倍率，觀察小鼠纖維母細胞L929貼附各金屬及合金之狀態 82
圖4-9 利用SEM以1,000倍率，觀察小鼠纖維母細胞L929貼附各金屬及合金之狀態 83
圖4-10小鼠纖維母細胞L929接觸各金屬及合金周圍之狀態 84
圖4-11小鼠纖維母細胞L929接觸各金屬及合金下方之狀態 85
圖4-12各金屬及合金浸泡於萃取培養基1天之狀態 86
圖4-13萃取1天培養基刺激細胞，經由MTT反應後的細胞染色狀態 87
圖4-14 各金屬及合金浸泡於萃取培養基3天之狀態 88
圖4-15萃取3天培養基刺激細胞，經由MTT反應後的細胞染色狀態 89
圖4-16 各金屬及合金浸泡於萃取培養基5天之狀態 90
圖4-17萃取5天培養基刺激細胞，經由MTT反應後的細胞染色狀態 91
圖4-18 Zr42Cu42Al8Ag8、Zr48Cu36Al8Ag8及Ti40Zr10Cu36Pd14無鎳金屬玻璃1、3、5天MTT assay定量分析結果 92
圖4-19 Zr42Cu42Al8Ag8、Zr48Cu36Al8Ag8及Ti40Zr10Cu36Pd14無鎳金屬玻璃檢測1、3、5天萃取培養基金屬離子溶出濃度 92
圖4-20 Zr42Cu42Al8Ag8、Zr48Cu36Al8Ag8及Ti40Zr10Cu36Pd14無鎳金屬玻璃檢測1、3、5天萃取培養基銅離子溶出濃度 93

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指導教授

鄭憲清(Shian-ching Jang)

審核日期

2014-7-29

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