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研究生:楊嘉芬
研究生(外文):Chia-fen Yang
論文名稱:探討微米尺度之鍍金粒幾丁聚醣膜表面對3T3纖維母細胞的貼附與生長的反應
論文名稱(外文):Study of the micro-scale Au deposited chitosan film surface responding to the attachment and proliferation of 3T3 fibroblast cells
指導教授:廖峻德廖峻德引用關係
指導教授(外文):Jiunn-der Liao
學位類別:碩士
校院名稱:國立成功大學
系所名稱:材料科學及工程學系碩博士班
學門:工程學門
學類:材料工程學類
論文種類:學術論文
畢業學年度:96
語文別:中文
論文頁數:70
中文關鍵詞:奈米尺度機械性質纖維母細胞蒸鍍上金群幾丁聚醣
外文關鍵詞:Chitosan filmnano-mechanicalfibroblast cellevaporated Au cluster
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幾丁聚醣由於具有無毒性、生物可降解性、生物相容性和價格便宜等優點,被視為優越的可運用於組織工程的生醫材料。雖然幾丁聚醣是一合適的支架材料,但對纖維母細胞幾乎沒有促進生長的效果。此外,由於細胞形貌與細胞和表面的作用息息相關,培養纖維母細胞在幾丁聚醣上,細胞型態多呈球狀。而細胞受生物基材表面的物理化學訊號影響,主要可分為三類:化學、形貌和機械性質,本研究以修飾特別製備的幾丁聚醣膜表面性質來改善細胞親和性。以電子束蒸鍍技術搭配蒸鍍罩,於幾丁聚醣膜表面蒸鍍上有圖案狀的微尺度金群。蒸鍍上的金群直徑約150 μm,厚度約20 nm。在幾丁聚醣膜上金群彼此間距1000 μm。以掃描式電子顯微鏡和共軛焦 3D 光學表面形貌量測儀來分析表面形貌。能量散佈光譜儀、光電子化學光譜儀和接觸角量測儀來分析化學性質。以薄膜壓痕試驗機的動態接觸模組作金群和幾丁聚醣基材的奈米尺度機械性質分析。實驗結果證明相較於幾丁聚醣基材,蒸鍍上的金群和金群附近的區域,具有較合適的聯合性質,去促進纖維母細胞的貼附性、攤開和生長狀況。且可觀察到細胞多聚集在此金群和金群附近的區域上。特別的是奈米尺度的表面機械性質在這些區域都有增進的效果。由金群造成的結合適當的化學及機械性質,在幾丁聚醣膜上創造一適宜纖維母細胞選擇性生長的表面,期望在組織工程的應用上可在引導細胞生長的影響多提供一種可能的方法。
Chitosan is classified as an excellent biomaterial owing to the advantages of non-toxicity, biodegradation, biocompatibility, and low cost. Though chitosan is regarded as a suitable scaffold material, it shows almost no promotion effect on the growth of cultured fibroblast cells. Besides, cell shapes induced by cell–surface interactions tend to become round. Cells on a biomaterial surface usually respond to three main categories of physicochemical cues: chemically, topographically, and mechanically. In this study, mechanical property of a particularly-prepared chitosan film was modified to improve its cell affinity. Micro-scale Au clusters were patterned and seeded on the chitosan film using a shadow mask and electron beam evaporator. The dimension of the evaporated Au cluster was 150 μm in diameter and 20 nm in thickness. They were separated at a distance of 1000 μm on the chitosan film. Scanning Electron Microscope and 3D Confocal Microscope were used to characterize their surface morphologies. X-ray Photoelectron Spectroscopy, Energy Dispersive Spectrometer, and water contact angles were employed to examine their chemical structures. Nano-indenter with dynamic contact module was applied to measure their nano-mechanical properties surrounding Au cluster and the chitosan matrix. Experimental results demonstrated that as compared with the chitosan matrix, the patterned Au clusters and their neighboring area exhibited a suitable and combined property to promote cell adhesion, spreading, and growth. Fibroblast cells were found accumulated surrounding Au clusters and their neighboring area. In particular, nano-mechanical properties in this domain significantly enhanced. In combination with the possible chemical and mechanical changes caused by Au cluster, a selectively-enhanced chitosan surface for fibroblast cells was created, which is promising for enabling a cells-guidable effect in tissue engineering.
第一章 序論 7
1.1 前言 7
1.2 研究動機 8
1.3 文獻回顧 8
1.3.1 生物支架基材 8
1.3.2 生物基材表面對細胞的影響 10
1.3.3 生物基材之表面修飾方法 13
1.4 研究大綱和目的 14
第二章 理論基礎 15
2.1 生物基材:幾丁質/幾丁聚醣 15
2.1.1幾丁質/幾丁聚醣簡介 15
2.1.2幾丁質/幾丁聚醣之生物應用 16
2.1.3 幾丁聚醣之力學性質 17
2.2基材表面修飾技術-物理氣相沉積(Physical Vapor Deposition, PVD) 18
第三章 實驗方法與設備 20
3.1 實驗藥品 20
3.2 實驗流程 21
3.3 實驗步驟 22
3.3.1製備幾丁聚醣膜 22
3.3.2 製備蒸鍍罩 22
3.3.3 表面修飾:於幾丁聚醣膜上以電子束蒸鍍機鍍上圖案狀的金薄膜層 22
3.3.4 培養3T3纖維母細胞於基材上 24
3.4 實驗儀器 25
3.4.1 二氧化碳雷射雕刻機(CO2 Laser Micro-machining System) 25
3.4.2 電子束蒸鍍機 ( E-beam Evaporator ) 26
3.4.3 掃瞄式電子顯微鏡(Scanning Electron Microscope, SEM) 28
3.4.4 靜態接觸角量測儀 30
3.4.5 奈米薄膜材料試驗機 (MTS Nano-indentator ) 31
3.4.6 化學分析電子光譜儀 (Electron Spectroscopy for chemical analysis, ESCA ) 34
第四章 結果與討論 35
4.1 以電子束蒸鍍技術對幾丁聚醣薄膜鍍金 35
4.1.1表面形貌分析 35
4.1.2化學元素分析 40
4.1.3 表面親、疏水性分析 46
4.1.4機械性質分析 48
4.2 生物性分析:培養3T3纖維母細胞 52
4.2.1 細胞形貌分析 52
4.2.2 細胞成長分析 56
4.2.3 細胞定點定時分析 60
第五章 結論 62
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