臺灣博碩士論文加值系統

本研究係利用共沉法來製備合成鋅摻雜的氫氧基磷灰石，目的是希望讓人工
骨骼材料具有抗菌能力。實驗中以添加不同濃度的氯化鋅來進行共沉法，以TGA檢測熱重損失，以熱裂解溫度分階段制定煅燒溫度。利用XRD檢測，實驗結果顯示經過煅燒後，發現添加氯化鋅濃度0.05 M以上時會出現些微氧化鋅與磷酸鋅鈣的新相。添加氯化鋅濃度到達0.15 M時，新相強度高過氫氧基磷灰石相成為主要存在的相。進行定性抗菌測試方法中的抑菌圈產生法測試，實驗中添加氯化鋅濃度由0.0125 M增加至0.15 M，結果顯示抗菌性隨鋅含量提高而上升。為模擬人工骨骼，以火焰噴塗法將過篩造粒後的粉末噴塗至Ti-6Al-4V基材上，進行平面及橫截面SEM觀察，發現鋅摻雜氫氧基磷灰石粉末以多孔結構方式存在，孔隙率明顯提升。進行抗菌測試，發現試片具有良好的抗菌性，抑菌率隨著試片與菌液的接觸時間增加而提升。進行細胞測試的生物活性及毒性測試，發現試片具有良好的生物相容性。

In this study, we prepared zinc-doped hydroxyapatite by the co-precipitation method, the purpose is to make artificial bone material has ability of antibacterial. The co-precipitation method prepared hydroxyapatite with add different concentrations of zinc chloride. Detected the weight loss by TGA to determine the sintering temperature of the pyrolysis temperature.Detected by XRD, the experimental results show that after sintering, it has some new phases generated of ZnO and CaZn2 (PO4)2, and the strength of new phases will high over than hydroxyapatite phase to be the major presence when we add zinc chloride that concentration reaches 0.15M. Testing antibacterial ability by the inhibition zone method of qualitative antibacterial test methods, the results shows that ability of antibacterial increased when the concentration of zinc chloride increased from 0.0125M to 0.15M. For simulate the artificial bone, the granulated powder that after sieved prilling were coated on the surface of Ti-6Al-4V substrate by the flame-spraying method, the SEM of surface topography and cross-section found that the zinc-doped hydroxyapatite was the porous structure on the substrate, and the porosity improved significantly. The antibacterial test found the specimen has a good antibacterial ability, the inhibition rate will increase when the time contacting by the specimen and bacteria increased. For biological activity and toxicity test of the cell test, the specimen was found with good biocompatibility.

摘 要 i
ABSTRACT ii
致 謝 iv
目 錄 v
表 目 錄 viii
圖 目 錄 ix
第一章 前言 1
1.1 研究背景 1
1.2 實驗動機與研究目的 1
第二章 理論基礎與文獻回顧 3
2.1 生醫材料 ( Biomaterials ) 3
2.1.1 生醫陶瓷之種類 ( Bioceramics ) 4
2.2 氫氧基磷灰石 ( Hydroxyapatite ) 5
2.2.1 氫氧基磷灰石之結構 8
2.2.2 氫氧基磷灰石之性質 9
2.2.3 氫氧基磷灰石之合成方法 11
2.3 鋅離子摻雜之影響 12
2.3.1 元素添加及取代對氫氧基磷灰石的影響 12
2.3.2 鋅離子之抗菌機構 13
2.4 披覆氫氧基磷灰石塗層之方法 15
2.4.1 火焰噴塗法 ( Flame Spraying ) 15
2.5 細菌培養與抑菌檢測技術之簡介 18
2.5.1 細菌生長研究方法 18
2.5.2 抑菌檢測技術 21
2.6 生物評估方法 24
2.6.1 細胞生物相容性 ( Biocompatibility ) 24
2.6.2 WST-1 細胞活性測試 25
2.6.3 LDH 細胞毒性測試 26
第三章 材料製備與實驗方法 27
3.1 實驗藥品及儀器 27
3.1.1 實驗藥品 27
3.1.2 實驗儀器 28
3.2 實驗方法與流程 30
3.2.1 實驗流程整體規劃 30
3.2.2 以共沉法製備氫氧基磷灰石粉末 31
3.2.3 以火焰噴塗法製備試片 34
3.3 材料性質分析 37
3.3.1 X光繞射儀分析 ( X-ray Diffractometer, XRD ) 37
3.3.2 熱重分析儀分析 ( Thermogravimetric Analyzer, TGA ) 38
3.3.3 掃描式電子顯微鏡 ( Scanning Electron Microscope ) 38
3.3.4粗糙度量測 ( Surface Roughness ) 39
3.3.5 孔隙率分析 ( Porosity ) 40
3.4 抗菌性分析 41
3.4.1 細菌實驗前置作業 41
3.4.2 定性抗菌 42
3.4.3 定量抗菌 43
3.5 生物相容性測試 45
3.5.1 細胞培養 45
3.5.2 WST-1細胞活性測試 47
3.5.3 LDH 細胞毒性測試 ( Cytotoxicity ) 48
第四章 結果與討論 49
4.1 粉末性質分析 49
4.1.1 XRD繞射峰分析 49
4.1.2 TGA熱重分析 52
4.1.3 SEM顯微組織觀察 54
4.1.4 EDS元素鑑定與半定量分析 56
4.1.5 定性抗菌性分析 57
4.2 火焰噴塗參數測試 59
4.2.1 XRD繞射峰分析 59
4.2.2 SEM顯微組織觀察 60
4.3火焰噴塗試片性質分析 64
4.3.1 XRD繞射峰分析 64
4.3.2 SEM顯微組織觀察 65
4.3.3 孔隙率計算 74
4.3.4 定性/量抗菌性分析 75
4.4 生物相容性評估 77
4.4.1 WST-1細胞活性測試 77
4.4.2 LDH細胞毒殺測試 78
第五章 結論 79
參考文獻 81

[1]Joon Bu Park, Biomaterials science and Engineering, New York: Plenum Press, 1984.
[2]Antony H. Melcher, "Summary of biological considerations," Journal of Dental Education, 1988, 52, 812.
[3]Larry L. Hench, "Bioceramics: from concept to clinic", Journal of American Ceramic Society, 1991, 74, 1487.
[4]Joon Bu Park and Roderic S. Lakes, Biomaterials an introduction, New York: Plenum Press, 1992.
[5]Vacanti C. A., Vacanti J. P., Bone and cartilage reconstruction of Tissue Engineering, New York: R.G. Landes Co, 1997, 619.
[6]Klaas de Groot, Bioceramics of calcium phosphate, Florida: CRC Press Inc., 1983.
[7]M. Jacho, "Calcium phosphate ceramics as hard tissue prosthetics," Clinical Orthopaedics and Related Research, 1981, 157, 259.
[8]Klaas de Groot, "Medical applications of calcium phosphate bioceramics," 日本協會學術論文誌, 1991, 99, 943-953.
[9]Joon Bu Park, Biomaterials Science and Engineering, New York: Plenum Press, 1985.
[10]鍾仁傑，以溶膠-凝膠法製備含鋅及銀之氫氧基磷灰石與其抗菌性質研究，碩士論文，清華大學材料科學工程研究所，新竹，2000。
[11]Joseph D.Bronzino, The Biomedical Engineering Handbook, Florida: CRC Press Inc., 1995.
[12]E. J. Duff and A. A. Grant, Mechanical Properties of Biomaterials, New York: John Wiley & Sons Ltd., 1980, 465.
[13]Antonio Ravaglioli and A. Krajewski, Bioceramics, London: Chapman＆Hall, 1992.
[14]董國忠，低鈣磷莫耳比磷酸鈣系玻璃之結晶性及物性研究，碩士論文，中原大學化學研究所，桃園，1996。
[15]Taki Negas and R. S. Roth, "Journal of Research of the National Bureau of Standards-A," Physics and chemistry, 1968, 72A(6), 783.
[16]H. Newesely and J. F. Osborn, Mechanical Properties of Biomaterials, New York: John Wiley & Sons Ltd., 1980, 465.
[17]A.C. Tas, F. Korkusuz, M. Timicin, Nuri Akkas, "An investigation of the chemical synthesis and high-temperature sintering behaviour of calcium hydroxyapatite (HA) and tricalcium phosphate (TCP) bioceramics," Journal of Materials Science-Materials in Medicine, 1997, 8(2), 91.
[18]Hua-Shan Liu, Tsung Shune Chin, L.S. Lai, S.Y. Chiu, K.H. Chung, C.S. Chang, M.T. Lui, "Hydroxyapatite synthesized by a simplified hydrothermal method," Ceramics International, 1997, 23(1), 19.
[19]Koji Ioku, Giichiro Kawachi, Manami Toda, Shuji Sasaki,Hirotaka Fujimori and Seishi Goto, "Porous Apatite Ceramics Prepared by Hydrothermal Method," 14th International Conference on the Properties of Water and Steam, Kyoto, 2004, 259-263.
[20]B. Ben Nissan, Adriyan Milev and Razi Vago, "Morphology of sol-gel derived nano-coated coralline hydroxyapatite," Biomaterials, 2004, 25, 4971-4975.
[21]J.S. Sun, H.C. Liu, Walter H.S. Chang, J. Li, F.H. Lin, H.C. Tai, "The influence of hydroxyapatite particle size on bone cell activities: an in vitro study," Journal Biomedical Materials Research, 1998, 39, 390.
[22]Mohammad R. Saeri, Abdollahe Afshar, Mohammad Ghorbani, Naser Ehsani and Charles Chris Sorrell, "The wet precipitation process of hydroxyapatite," Materials Letters, 2003, 57, 4064-4069.
[23]Johanna M. Jarcho, C.H. Bolen, M. B. Thomas, J. Bobick, J. F. Kay, Robert H. Doremus, "Hydroxylapatite synthesis and characterization in dense polycrystalline form," Journal of Materials Science, 1976, 11(11), 2027-2035.
[24]J. Zhou, X. Zhang, J. Chen, S. Zeng, Klaas de Groot, "High temperature characteristics of synthetic hydroxyapatite," Journal of Materials Science-Materials in Medicine, 1993, 4(1), 83-85.
[25]Michael J. Root, "Inhibition of the amorphous calcium phosphate transformation reaction by polyphosphonates and metal ions," Calcified Tissue International, 1990, 47, 112.
[26]Y. Okamoto and Sumitomo Hidaka, "Studies on calcium phosphate precipitation : Effects of metal ions used in dental materials," Journal of Biomedical Materials Research, 1994, 28, 1403.
[27]A. Bigi, Eugenio Foresti, Maria Gandolfi, Massimo Gazzano and N. Rovri, "Inhibiting effect of Zinc on hydroxyapatite crystallization," Journal of Inorganic Biochemistry, 1995, 58, 49.
[28]N. Kanzaki, K. Onuma, G. Treboux, S. Tsutsumi and Atsuo Ito, "Inhibitory effect of magnesium and zinc on crystallization kinetics of hydroxyapatite (0001) face," Journal of Physical Chemistry B, 2000, 104, 4189.
[29]Atsuo Ito, K. Ojima, H. Naito, N. Ichinose and T. Tateishi, "Preparation, solubility, and cytocompatibility od zinc-releasing calcium phosphate ceramics," Journal of Biomedical Materials Research, 2000, 50, 178.
[30]H. Kawamura, Atsuo Ito, S. Miyakawa, P. Layrolle, K. Ojima, N. Ichinose, T. Tateishi, "Stimulatory effect of zinc-releasing calcium phosphate implane on bone formation in rabbit femora," Journal of Biomedical Materials Research, 2000, 50, 184.
[31]Baljit S. Moonga and David W. Dempster, "Zinc is a potent inhibitor of osteocalstic bone resorption in vitro," Journal of Bone and Mineral Research, 1995, 10(3), 453.
[32]王鏡岩，朱聖庚，徐長法，生物化學，北京：高等教育出版社，2002，第474頁。
[33]S. K. Yen, C. M. Lin, "Cathodic reactions of electrolytic hydroxyapatite coating on pure titanium," Material chemistry and phys, 2002, 77, 70.
[34]C. F. Feng, K. A. Khor, E. J. Liu, P. Cheang, "Phase transformations in plasma sprayed hydroxyapatite coatings," Scripta Material, 2000, 42, 103.
[35]Yifei Hu, Xigeng Miao, "Comparson of hydroxyapatite ceramics and hydroxyapatite/borosilicate glass composites prepared by slip casting," Ceramic international, 2004, 30, 1788.
[36]Miguel Manso, Samuel Ogueta, Pilar Herrero-Fernandez, Luis Vazquez, Michel Langlet, Josefa Predestinacion Garcia-Ruiz, "Biological evaluation of aerosol-gel-derived hydroxyapatite coatings with human mesenchymal stem cells," Biomaterials, 2002, 23, 3985.
[37]Lech Pawlowski, 熱噴塗科學與工程(The science and engineering of thermal spraycoatings)，北京：機械工業出版社，2011，第36頁。
[38]王娟，表面堆焊與熱噴塗技術，北京：化學工業出版社，2004，第119-120頁。
[39]American association of textile chemists and colorists, "Antibacterial Activity of Fabrics, Detection of: Agar Plate Method," ATCC Test Method 90-1982.
[40]American association of textile chemists and colorists, "Antibacterial Activity Assessment of Textile Materials: Parallel Streak Method," AATCC Test Method 147-1998.
[41]American national standards institute, "Practice for Determining Resistance of Plastics to Bacteria," ASTM G22-76.
[42]American national standards institute, "Standard Test Method for Determining the Antimicrobial Activity of Immobilized Antimicrobial Agents Under Dynamic Contact Conditions," ASTM E2149-01.
[43]Japanese Industrial Standard, "Antimicrobial products : Test for antimicrobial activity and efficacy (foreign standard)," JIS Z 2801: 2000.
[44]T. Mosmann, "Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays," J Immunol Methods, 1983, 65, 55-63.
[45]Neal W. Roehm, G. H. Rodgers, S. M. Hatfield, and Andrew L. Glasebrook, "An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT," Journal of Immunological Methods, 1991, 142, 257-265.
[46]H. Cory, T. C. Owen, J. A. Barltrop, and J. G. Cory, "Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture," Cancer communications, 1991, 3, 207-212
[47]M. Jarcho, J. F. Kay, K. I. Gumaer, R. H. Doremus, and H. P. Drobeck, "Tissue, cellular and subcellular events at a bone-ceramic hydroxylapatite interface," Journal of Bioscience and Bioengineering, 1977, 1, 79-92.
[48]盧光舜，消毒學，南山堂出版社，台北，1985。
[49]J. Suwanprateeb, K. E. Tanner, S. Turner, and W. Bonfield, "Influence of sterilization by gamma irradiation and of thermal annealing on creep of hydroxyapatite-reinforced polyethylene composites," Journal of Biomedical Material Research, 1998, 39, 16-22.
[50]林麗娟，「X光繞射原理及其應用」，工業材料，第八十六期，1994，第100-109頁。
[51]謝飛，曹有名，雷周橋，周心其，雷敏娟，「超細磷酸鋅鈣粉體的製備與性能」，中南大學學報(自然科學版)，第四十三卷，第四期，2012，第1295-1298頁。