臺灣博碩士論文加值系統

目前常用的人工膝關節股骨與脛骨基座部分以及膝關節之材料主要為鈷鉻鉬或鈦合金，而其兩者接合處之半月板則為超高分子量聚乙烯（UHMPE）所組成；這種的人工關節植入人體後很容易因為磨損產生許多碎屑，使得植入人工膝關節的患者發生骨質溶解（osteolysis）現象，造成人工關節鬆脫，進而導致手術的失敗。
過去對於模擬磨耗顆粒的研究，使用的材料多以純金屬顆粒為主，與實際的人工關節材料成份上有一段差距，本研究使用旋轉電極法自製鈷鉻鉬合金顆粒，加上純金屬鈷、鎳顆粒和鈷、鎳二價離子化合物，以J774鼠巨噬細胞做測試，探討生物相容性。
研究結果發現，自製鈷鉻鉬合金顆粒、純金屬鈷、鎳顆粒和鈷、鎳正二價離子皆會對細胞造成毒性，但自製鈷鉻鉬合金顆粒的毒性比純金屬鈷、鎳顆粒低，且鈷鉻鉬合金顆粒、純金屬鈷、鎳顆粒在溶液中會釋放離子，推測金屬顆粒造成細胞死亡原因包含釋放的離子造成的毒性。
本研究成功以旋轉電極法自製鈷鉻鉬合金顆粒，模擬人工關節磨耗產生的顆粒，對於日後相關研究，可提供一個不同的實驗方向。

Most of the total joint prosthesis used currently are made of metal base and ultra-high molecular weight polyethylene （UHMWPE）. Wear debris from UHMWPE （found in the artificial joint implants） may easily cause osteolysis, which may lead to the loosening of the implant and, hence, failure of prosthesis.
The materials used in the researches in the past on simulating wear particles from the joint prosthesis were mostly pure metal particles, but there were a lot of differences between the real compositions of joint prosthesis and pure metal particles. This study used the Co-Cr-Mo alloy particles created by rotational electrode process, cobalt, nickel pure metal particles and cobalt, nickel chemical compound as materials, and cultured J774 murine macrophage with these materials for examine the biocompatibility.
As a result, Co-Cr-Mo alloy particles created by rotational electrode process, cobalt, nickel pure metal particles and cobalt, nickel chemical compound all cause cytotoxicity to the cells, but the Co-Cr-Mo alloy particles were less cytotoxic to cobalt, nickel pure metal particles. And we found that cobalt, nickel pure metal particles would release ions in solutions, we supposed that the reason of the death of the cells made by metal particles includes the cytotoxicity created by the release of ions.
This study produced the Co-Cr-Mo alloy particles successfully by rotational electrode process, to simulate the wear particles from the joint prosthesis. It provides an alternative way to experiment in relative researches in the future.

中文摘要 1
英文摘要 2
縮寫對照表 4
第一章 緒論 6
1.1 生醫材料與人工關節材料的發展 6
1.1.1 生醫材料之研究與發展 6
1.1.2 人工關節材料之發展 7
1.1.3 常用金屬植入材料 8
1.1.4 生物相容性 11
1.1.5 生醫金屬材料的腐蝕與磨耗 14
1.2 生物反應 16
1.2.1 細胞死亡 16
1.2.2 鈷、鎳離子的毒性 17
1.2.3 磨耗顆粒誘導的發炎 19
1.2.4 活性氧 20
1.3 旋轉電極法原理及應用 21
1.4 研究動機 23
第二章 材料與實驗方法 25
2.1 測試材料 25
2.1.1 旋轉電極法自製鈷鉻鉬顆粒 25
2.2 掃瞄穿透式電子顯微鏡與能量分散光譜儀分析 27
2.3 原子吸收光譜儀 27
2.4 細胞培養 28
2.5 錐蟲藍排除法 28
2.6 細胞存活率試驗 28
2.7 Acridine Orange 染色細胞型態分析 29
2.8 Particle-soaked solution試驗 29
2.9 統計分析 30
第三章 實驗結果 31
3.1 生醫材料顆粒之型態與成份分析 31
3.2 金屬顆粒和離子造成J774細胞死亡 31
3.3 EDTA可抑制細胞因金屬顆粒或離子造成的死亡 32
3.4 Acridine Orange 染色細胞型態分析 33
3.5 Particle-soaked solution試驗及離子釋放量分析 33
第四章 討論 35
參考文獻 61

參考文獻
Mangel, A. W. L. Scott and Liddle, R. A. Depolarization-stimulated cholecystokinin secretion is mediated by L-type calcium, Am J Physiol Gastrointest Liver Physiol 270: G287-G290 (1996)
Alan Petit et al. Differential apoptotic response of J774 macrophages to alumina and ultra-high-molecular-weight polyethylene particles. J. Orthopaedic Research 20, 9-15 (2002)
Alan Petit et al. Effect of cobalt and chromium ions bcl-2, bax caspase-3, and caspase-8 expression in human U937 macrophages. Biomaterials 25, 2013-2018 (2004)
Andree, H.A. et al. Binding of vascular anticoagulant alpha(VAC alpha) to planar phospholipid bilayers. J Biol Chem 265, 4923-4928 (1990).
Boabaid, F., Cerri, P. S. & Katchburian, E. Apoptotic bone cells may be engulfed by osteoclasts during alveolar bone resorption in young rats. Tissue Cell 33, 318-325 (2001).
Charnley, J., Follacci, F. M. & Hammond, B. T. The long-term reaction of bone to self-curing acrylic cement. J. Bone Joint Surg. Br. 50, 822-829 (1968).
Charnley,J. & Cupic,Z. The nine and ten year results of the low-friction arthroplasty of the hip. Clin. Orthop. 95:9-25., 9-25 (1973).
Daniels,A.U., Barnes,F.H., Charlebois,S.J. & Smith,R.A. Macrophage cytokine response to particles and lipopolysaccharide in vitro. J. Biomed. Mater. Res. 49, 469-478 (2000).
Dobbs,H.S. & Minski,M.J. Metal ion release after total hip replacement. Biomaterials 1, 193-198 (1980).
Dobbs,H.S. Survivorship of total hip replacements. J. Bone Joint Surg. Br. 62-B, 168-173 (1980).
Ducheyne,P., Cohn,C.S.: Biomaterial: structure, processing, and mechanical properties, In: Steinberg MemEd. The hip and its disorders. WN Saunders, Philadelphia, 905-928 (1991).
Ellis,R.E., Yuan,J.Y., and Horvitz, H.R. Mechanisms and functions of cell death. [Review][175 refs]. Annual Review of Cell Biology.v.7, 663-698 (1991).
Ertel,S.I., Ratner,B.D., Kaul,A., Schway,M.B. & Horbett,T.A. In vitro study of the intrinsic toxicity of synthetic surfaces to cells. J Biomed. Mater. Res 28, 667-675 (1994).
Evans,E.J. Cell damage in vitro following direct contact with fine particles of titanium, titanium alloy and cobalt-chrome-molybdenum alloy. Biomaterials 15, 713-717 (1994).
Farrar,M.A. & Schreiber,R.D. The molecular cell biology of interferon-gamma and its receptor. Annu. Rev. Immunol 11:571-611., 571-611 (1993).
Fei Chen, Xianglin Shi. Intracellular signal transduction of cells in response to carcinogenic metals. Oncology hematology 43, 105-121 (2002).
Galante,J.O., Lemons,J., Spector,M., Wilson,P.D., Jr. & Wright,T.M. The biologic effects of implant materials. J Orthop. Res 9, 760-775 (1991).
Gawkrodger D.J. Metal sensitivities and orthopaedic implants revisited: thepotential for metal allergy with the new metal-on-metal joint prostheses. British Journal of Dermatology; 148: 1089–1093(2003).
Georgescu,H.I., Mendelow,D. & Evans,C.H. HIG-82: an established cell line from rabbit periarticular soft tissue, which retains the "activatable" phenotype. In Vitro Cell Dev. Biol. 24 , 1015-1022 (1988).
Glant,T.T. & Jacobs,J.J. Response of three murine macrophage populations to particulate debris: bone resorption in organ cultures. J. Orthop. Res. 12, 720-731 (1994).
Goldring,S.R., Clark,C.R., and Wright,T.M. Aseptic Loosening. The Journalof Bone and Joint Surgery (American Volume) 75-(A), 799-801(1993).
Gougeon,M.L., Lecoeur,H., Dulioust,A., Enouf,M.G., Crouvoiser,M., Goujard,C. Debord,T., Montagnier,L. Programmed cell death in peripheral lymphocytes from HIV-infected persons: increased susceptibility to apoptosis of CD4 and CD8 T cells correlates with lymphocyte activation and with disease progression. Journal of Immunology. 156, 3509-20(1996).
Green T. R., Fisher J., Stone M., Wroblewski B.M., Ingham E.. Polyethylene particles of a "critical size" are necessary for the induction Of cytokines by macrophages in vitro. Biomaterials 192297-2302Volume: 138, Issue: 1-3, July 20, 2003, pp. 231-235 (1998).
Hallab,N., Merritt,K. & Jacobs,J.J. Metal sensitivity in patients with orthopaedic implants. J Bone Joint Surg Am. 83-A, 428-436 (2001).
Harris,W.H., Schiller,A.L., Scholler,J.M., Freiberg,R.A. & Scott,R. Extensive localized bone resorption in the femur following total hip replacement. J. Bone Joint Surg. Am. 58, 612-618 (1976).
Hastings,G.W. Biomedical engineering and materials for orthopaedic implants. J Phys. [E. ] 13, 599-607 (1980).
Haynes,D.R., Rogers,S.D., Hay,S., Pearcy,M.J. & Howie,D.W. The differences in toxicity and release of bone-resorbing mediators induced by titanium and cobalt-chromium-alloy wear particles. J Bone Joint Surg Am. 75, 825-834 (1993).
Haynes,D.R., Boyle,S.J., Rogers,S.D., Howie,D.W. & Vernon-Roberts,B. Variation in cytokines induced by particles from different prosthetic materials. Clin. Orthop. 223-230 (1998).
Hibi,M., Lin,A., Smeal,T., Minden,A. and Karin,M. Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain. Genes and Development, 7, 2135-2148 (1993).
Howie,D.W. & Vernon-Roberts,B. Synovial macrophage response to aluminium oxide ceramic and cobalt-chrome alloy wear particles in rats. Biomaterials 9, 442-448 (1988).
Howie,D.W. & Vernon-Roberts,B. The synovial response to intraarticular cobalt-chrome wear particles. Clin. Orthop. 244-254 (1988).
Ingham,E. & Fisher,J. Biological reactions to wear debris in total joint replacement. Proc Inst. Mech. Eng [H. ] 214, 21-37 (2000).
Isabelle Catelas et al. TNF-a secretion and macrophage mortality induced by conalt and chromium ions in vitro-Qualitative analysis of apoptosis. Biomaterials 24, 383-391 (2003).
Jacobs,J.J., Roebuck,K.A., Archibeck,M., Hallab,N.J. & Glant,T.T. Osteolysis: basic science. Clin. Orthop. 71-77 (2001).
Jacobsson,S.A., Djerf,K., and Wahlstrom,O. Twenty-year results of McKee-Farrar versus Charnley prosthesis. Clin. Orthop. S60-S68 (1996).
Wang, J. Y. et al. Titanim, chromium and cobalt ions modulate the release of bone-associated cytokines by human monocytrs/macrophages in vitro. Biomaterials 17 2233-2240 (1996)
Jeppesen,C. & Nielsen,P.E. Photofootprinting of drug-binding sites on DNA using diazo- and azido-9-aminoacridine derivatives. Eur. J Biochem 182, 437-444 (1989).
Kawalec, J.S., Brown, S.A., Payer, J.H and Merritt, K. Mixed-metal fretting corrosion of Ti6Al4V and wrought cobalt alloy. Journal of Biomedical Materials Research. 29, 867-873 (1995).
Kerr, J.F., Wyllie, A.H., and Currie,A.R. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26, 239–257 (1972).
Kilner, T., Pilliar, R.M., Weatherly, G. C., and Allibert, C. Phase identification and incipient melting in a cast CoCr surgical implant alloy. Journal of Biomedical Materials Research 16, 63-79 (1982).
Kyungtae Kim et al. Nickel-induced apoptosis in murine T cell hybridoma cells is associated with increased Fas ligand expression. Toxicology and applied pharmacology 185, 41-47 (2002).
La Budde,J.K., Orosz,J.F., Bonfiglio,T.A. and Pellegrini,V.D. Jr. Particulate titanium and cobalt-chrome metallic debris in failed total knee arthroplasty. A quantitative histologic analysis. Journal of Arthroplasty 9, 291-304 (1994)
Linehan,A.D. & Windeler,A.S. Passive fit of implant-retained prosthetic superstructures improved by electric discharge machining. J Prosthodont. 3, 88-95 (1994).
Macklis,R.M., Lin,J.Y., Beresford,B., Atcher,R.W., Hines ,J.J., Humm, J.L.. Cellular kinetics, dosimetry, and radiobiology of alpha-particle radioimmunotherapy: induction of apoptosis. Radiation Research. 130, 220-226 (1992).
Maloney,W.J., Jasty,M., Harris,W.H., Galante,J.O. & Callaghan,J.J. Endosteal erosion in association with stable uncemented femoral components. J. Bone Joint Surg. Am. 72, 1025-1034 (1990).
Manolagas,S.C. Role of cytokines in bone resorption. Bone 17, 63s-67s (1995b).
Marisa D. Pulido, Alan R. Parrish. Metal-induced apoptosis: mechanism. Mutation Research 533, 227-241 (2003).
McConkey,D.J., Orrenius, S., and Jondal, M. Cellular signaling in programmed cell death (apoptosis). [Review][21 refs]. Immunology Today,v. 11, 120-121 (1990).
McKellop,H. et al. In vivo wear of three types of metal on metal hip prostheses during two decades of use. Clin. Orthop. S128-S140 (1996).
Morais,S., Sousa,J.P, Fernandes,M.H., Carvalho,G.S., de Bruijn,J.D., van, and Blitterswijk,C.A.. Effect of AISI 316L corrosion products in in vitro bone formation. Biomaterials 19, 999-1007 (1998).
Mosmann,T. - Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. - J Immunol Methods 1983 Dec 16;65(1-2):55-63. 55-63 (1983).
Mostardi,R.A., Meerbaum,S.O., Kovacik,M.W. & Gradisar,I.A., Jr. Response of human fibroblasts to tantalum and titanium in cell culture. Biomed. Sci. Instrum. 33:514-8., 514-518 (1997).
Mostardi,R.A., Meerbaum,S.O., Kovacik,M.W. & Gradisar,I.A., Jr. In vitro response of human fibroblasts to commercially pure titanium. J. Biomed. Mater. Res. 47, 60-64 (1999).
Mostardi,R.A., Pentello,A., Kovacik,M.W. & Askew,M.J. Prosthetic metals have a variable necrotic threshold in human fibroblasts: an in vitro study. J. Biomed. Mater. Res. 59, 605-610 (2002).
Muller,M.E. The benefits of metal-on-metal total hip replacements. Clin. Orthop. 54-59 (1995).
Olga L. Huk et al. Induction of apoptosis and necrosis by metal ions in vitro. J. Arthroplasty 19, 84-87 (2004).
Pan,J., Leygraf,C., Thierry,D. & Ektessabi,A.M. Corrosion resistance for biomaterial applications of TiO2 films deposited on titanium and stainless steel by ion-beam-assisted sputtering. J Biomed. Mater. Res 35, 309-318 (1997).
Pariza,M.W. Animal studies: summary, gaps, and future research. American Journal of Clincal Nutrition 66, 1539S-1540S (1997).
Prabhu,A., Shelburne,C.E. & Gibbons,D.F. Cellular proliferation and cytokine responses of murine macrophage cell line J774A.1 to polymethylmethacrylate and cobalt-chrome alloy particles. J. Biomed. Mater. Res. 42, 655-663 (1998).
Rae,T. A study on the effects of particulate metals of orthopaedic interest on murine macrophages in vitro. J Bone Joint Surg Br. 57, 444-450 (1975).
Rae,T. The haemolytic action of particulate metals (Cd, Cr, Co, Fe, Mo, Ni, Ta, Ti, Zn, Co-Cr alloy). J. Pathol. 125, 81-89 (1978).
Rae,T. Comparative laboratory studies on the production of soluble and particulate metal by total joint prostheses. Arch. Orthop. Trauma Surg. 95, 71-79 (1979).
Rae,T. The toxicity of metals used in orthopaedic prostheses. An experimental study using cultured human synovial fibroblasts. J Bone Joint Surg Br. 63-B, 435-440 (1981).
Regina,L.W., Messer, Sanford, Bishop, and Linda,C.L. Effect of metallic toxicity human gingival fibroblasts morphology. Biomaterials 20,1647-1657 (1999).
Gordon, S. The role of the macrophage in immune regulation. Res. Immunol. 149, 685-688 (1998).
Kim , S. H. and Sharma , R. P. Mercury-induced apoptosis and necrosis in murine macrophage: role of calcium-induced reactive oxygen species and p38 mitogen-activated protein kinase signaling. Toxicology and Applied Pharmacology 196, 47-57 (2004).
Semlitsch,M., Streicher,R.M. & Weber,H. [The wear behavior of capsules and heads of CoCrMo casts in long-term implanted all-metal hip prostheses]. Orthopade 18, 377-381 (1989).
Stea,S. et al. Apoptosis in peri-implant tissue. Biomaterials 21, 1393-1398 (2000).
Stefanovic-Racic,M., Stadler,J., Georgescu,H.I. & Evans,C.H. Nitric oxide synthesis and its regulation by rabbit synoviocytes. J. Rheumatol. 21, 1892-1898 (1994).
Stys P. K., Ransom B. R. and Waxman S. G., Effects of polyvalent cations and dihydropyridine calcium channel blockers on recovery of CNS white matter from anoxia. Neurosci. Lett. 115, 293-299 (1990)
Sung,K., Mendelow,D., Georgescu,H.I. & Evans,C.H. Characterisation of chondrocyte activation in response to cytokines synthesised by a synovial cell line. Biochim Biophys Acta 971, 148-156 (1988).
Sutula,L.C. et al. The Otto Aufranc Award. Impact of gamma sterilization on clinical performance of polyethylene in the hip. Clin. Orthop. 28-40 (1995).
Sychterz, C.J., Moon, K.H., Hashimoto, Y., Terefenko, K. M., Engh, C. A., Jr., and Bauer, T.W. Wear of polyethylene cups in total hip arthroplasty. A study of specimens retrieved post mortem. J Bone Joint Surg Am. 78, 1193-1200 (1996).
Hanawa , T. Metals ion release from metal implants. Materials science and engineering C24, 745-752 (2004).
Tang,L. & Eaton,J.W. Inflammatory responses to biomaterials. Am. J Clin. Pathol. 103, 466-471 (1995).
Tsuchiya T. Study on the standardization of cytotoxicity tests and new standard reference materials useful for evaluating the safety of biomaterials. Journal of Biomater 9, 138 (1994).
Engeland, M., Nieland, L. J., Ramaekers, F.C., Schutte, B. & Reutelingsperger,C.P. Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure. Cytometry 31, 1-9 (1998).
Wallace,L.,Guess,S., Alan, Rosenbluth et al. Agar difusion method for toxicyty screening of plastics on culture cell monolayers. Journal of Pharmaceutical Science. 54, 1545 (1965).
Wang,J.Y., Wicklund,B.H., Gustilo,R.B. & Tsukayama,D.T. Titanium, chromium and cobalt ions modulate the release of bone-associated cytokines by human monocytes/macrophages in vitro. Biomaterials 17, 2233-2240 (1996).
Weber,B.G., Semlitsch,M.F. & Streicher,R.M. Total hip joint replacement using a CoCrMo metal-metal sliding pairing. Nippon Seikeigeka Gakkai Zasshi 67, 391-398 (1993).
Willert,H.G. Reactions of the articular capsule to wear products of artificial joint prostheses. J. Biomed. Mater. Res. 11, 157-164 (1977).
Williams,D.F. Biomaterials and biocompatibility. Med Prog. Technol. 20;4, 31-42 (1976).
Wong,A.C. et al. Calcinosis circumscripta. A case report. J. Bone Joint Surg. Am. 68, 297-299 (1986).
Wright,T.M., Rimnac,C.M., Stulberg,S.D., Mintz,L., Tsao,A.K., Klein,R.W., and McCrae, CWear of polyethylene in total joint replacements. Observations from retrievedPCA knee implants. Clinical Orthopaedics & Related Research 276, 126-134. (1992).
Wyllie,A.H. Glucocorticoid-induced thymocyte apoptosis is as-sociated with endonuclease activation. Nature (London) 284,555–556 (1980).
Yasuo Nikia, Hideo Matsumotoa, Yasunori Sudaa, Toshiro Otania, Kyosuke Fujikawab,Yoshiaki Toyamaa, Noriyuki Hisamoric and Akira Nozue, Metal ions induce bone-resorbing cytokine production through the redox pathway in synoviocytes and bone marrow macrophages. Biomaterials 24 , 1447–1457 (2003)
Zuoning Han, David L. Boyle, Lufen Chang, Brydon Bennett, Michael Karin, Li Yang, Anthony M. Manning, and Gary S. Firestein, c-Jun N-terminal kinase is required for metalloproteinase expression and joint destruction in inflammatory arthritis, J. Clin. Invest. 108:73–81 (2001).
朱峰民，人工關節材料顆粒引發兔滑液膜細胞凋亡作用機制之研究，台灣大學，碩士論文，2004年1月。
陳誌賢，旋轉電極法製造FeAlNd合金粉末之磁性研究，台灣大學，碩士論文，2004年6月。
黃坤祥，粉末冶金學，中華民國粉末冶金協會，2001年1月。
雍喆婷，人工關節磨耗顆粒引發兔滑液膜細胞凋亡和吞噬作用之研究，台灣大學，碩士論文，2002年6月。