The study of metal particles and releasing ions inducing cell death on J774 macrophages

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 6