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研究生:楊登翔
研究生(外文):Teng-hsiang Yang
論文名稱:週期拉伸對造骨細胞的細胞型態的影響
論文名稱(外文):Cylic stretch Effects on Cell Morphology of Osteoblast cells
指導教授:蘇芳慶蘇芳慶引用關係
指導教授(外文):Fong-Chin Su
學位類別:碩士
校院名稱:國立成功大學
系所名稱:醫學工程研究所碩博士班
學門:工程學門
學類:綜合工程學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:58
中文關鍵詞:造骨細胞機械應力細胞力學排列型態
外文關鍵詞:osteoblastsmechanical forcecell mechanicsorientationmorphology
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骨組織為提供骨骼機械性質與新陳代謝功能的礦化組織,在骨組織中含有三種細胞:造骨細胞、骨細胞與蝕骨細胞。過去有許多研究證實骨組織在受到機械刺激時,細胞會因應機械刺激而改變細胞骨架的組成調節其型態與排列方向適應刺激。
本研究主要目的在於探討造骨細胞的型態變化與機械刺激間的關係。實驗中選用 MG-63 人的造骨細胞,培養在塗布纖維網蛋白的彈性薄膜 24 小時候,分別施加單軸向 5%,1 赫茲、0.5 赫茲、0.17 赫茲與 10%,1 赫茲、0.5 赫茲、0.17 赫茲的刺激,實驗時間為 120 分鐘,每 30分鐘觀察一次,並量化細胞的面積、長度、寬度與排列角度。
實驗結果顯示,細胞受到刺激時,面積與寬度都會隨著實驗的進行而減少,並且其減少的速度會受到拉伸應變的影響;細胞排列也隨著實驗的進行而趨近垂直於拉伸軸向,並且拉伸應變愈大其排列角度愈大。另外對細胞施加雙軸向的拉伸,並在兩個軸向施加不同的拉伸應變刺激 30分鐘。結果顯示:細胞會垂直於應變量較大的軸向排列。
Bone tissue is the mineralized tissue which provided multiple mechanical property and metabolic functions to the skeleton. The bone cells include osteoblasts, osteocytes and osteoclasts. When the bone is subject to the mechanical stimulation, the cell may adjust the morphology and reorient to a particular direction.
In this study, the relationship between cyclic stretch and cell morphology was investigated. We cultured osteoblast cell, MG-63 on the isotropic silicon chamber, which pre-coated fibronectin, and applied uni-axial stretch with 10% strain ratio at 1Hz, 0.5Hz, and 0.17Hz as well as 5% at 1Hz, 0.5Hz, and 0.17Hz, for 120 min respectively. The cell morphology was recorded every 30min by a CCD camera and analyzed by image software. The cell spreading area, length, width, and alignment angle were measured and analyzed.
The results showed that the cell spreading area and width decreased with time, and the decreased rate was influenced by strain ratio. The cell orientation was perpendicular to the stretching direction. The alignment angle between cell’s major axis and stretch axis increased with increasing strain ratio. In addition, we applied the bi-axial stretch to cell with different strain ratio in two axes for 30min. We found that cell orientation was perpendicular to the direction with maximal substrate deformation.
中文摘要..................I
Abstract.............................II
致謝...........................III
目錄..............................IV
表目錄..............................VII
圖目錄...............................VIII
第一章 緒論...............................1
1.1 骨組織...........................1
1.1.1 造骨細胞........................2
1.1.2 細胞骨架.................3
1.2 機械刺激.....................4
1.2.1 機械刺激對造骨細胞的影響............4
1.2.2 細胞型態的變化與機械拉伸的關係......9
1.3 實驗動機與目的.......11
第二章 實驗方法.............12
2.1 細胞培養.............12
2.2 實驗儀器.....13
2.2.1顯微鏡工作平台...........13
2.2.2 細胞拉伸系統.......15
2.2.3 溫度控制系統............19
2.3 影像處理與分析............21
2.4 統計分析..........23
2.5 實驗設計...............24
2.6 實驗流程........25
第三章 結果............26
3.1 細胞在週期拉伸中的型態表現............26
3.2 拉力產生機測試.................28
3.3 拉伸刺激下細胞面積的變化化.............33
3.4 細胞寬度的變化............36
3.5 細胞延伸量變化..........39
3.6 細胞排列角度.............42
3.7 雙軸不等量拉伸對細胞型態的影響.........47
第四章 討論...........49
4.1 實驗結果的討論............49
4.1.1 細胞型態的變化....................49
V 4.1.2 細胞排列的變化..............50
4.1.3 雙軸不等量拉伸的影響...........51
4.2 細胞骨架與細胞型態.........52
4.3 實驗限制因素............54
4.3.1 細胞密度的影響...........54
4.3.2 儀器限制.............54
4.3.3 影像處理造成的實驗誤差......55
第五章 結論與未來展望.........56
參考文獻..............57
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