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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/65239
標題: | 氣壓轉注成型製作三維生醫支架於組織工程之應用 Pneumatic Transfer Molding to Fabricate Three-Dimensional Biodegradable Porous Scaffolds |
作者: | MING-TUNG Lin 林銘棟 |
指導教授: | 楊申語(Sen-Yeu Yang) |
關鍵字: | 氣壓轉注加壓成型,孔隙率,吸水率,生醫支架,PDMS,細胞,軟骨, Pneumatic Transfer Molding,Porosity,Mositure,Scaffold,PDMS,Cell,Cartilage, |
出版年 : | 2012 |
學位: | 碩士 |
摘要: | 多孔性3D生醫支架主要運用於組織工程中,利用自體之組織細胞進行體外培養,再經由生長因子或訊號的刺激,使細胞生長分化成所需之組織形貌,待組織生長後植回體內,達到組織修復之目的。
現今製作支架的方式,大多採用有機溶劑或快速成型等方式製作,前者有溶劑揮發不完全問題;後者有無法一體成型,需使用電腦複雜程序控制一層一層堆積成所需孔洞支架。本研究開發氣壓轉注加壓成型機,特點為不使用任何有機溶劑,即可製作3D具孔洞支架,且可一體成形,可快速製作支架。本研究在製作圓盤型生醫支架前,先行進行最佳混合比測試,擇出以PLGA與NaCl重量比1:11有最佳支架性質。接著,進行尋求最佳成型參數與探討其支架機械性質,實驗結果其最佳成形參數為模溫90℃、料管加熱溫度200℃,壓力為4 kgf/cm2,製作出支架其孔隙率90%以上,孔洞大小為200~500 μm,符合其生醫用軟骨細胞生長大小。另外,本研究也進行縮小版人耳形狀支架製作,以材質PDMS作為軟模進行測試,實驗證實可製作出具3D孔洞耳朵型支架,孔洞大小200~500 μm。 圓盤型支架製作後,進行支架降解性測試,及實際細胞生長於支架上,結果顯示,於定性檢測中,使用SEM及螢光共焦顯微儀觀察細胞生長情形,皆有明顯貼覆及生長於支架內部;於定量檢測中,細胞基因表現、MTT活性檢測,皆顯示隨培養天數增加,細胞於支架適應性提高,細胞生長量與細胞外基質含量也隨之增加,MTT試劑檢測於一天時,有84%之軟骨細胞生長率,顯示軟骨細胞對於此支架環境適應良好。 Porous 3D biomedical scaffolds have been widely used in tissue engineering. The scaffold can be used to culture autologous cells in vivo, stimulated by growth factors and signals. The cells can grow into the desired tissue morphology. Finally, it can be transplanted back into the body, through the stable tissue growth achieving the purpose of tissue repair. Conventional scaffolds are mostly prepared by organic solvents or rapid prototyping. There are problems such as residual solvent and complicated process. In order to solve these problems, a pneumatic transfer molding machine is designed and implemented to fabricate scaffolds in this study. We first mold the disk biomedical scaffolds. It is found that the best weight ratio of PLGA:NaCl is 1:11. The experimental result also shows the best forming parameters are 90℃ for mold temperature, 200℃for the tube heating temperature and 4 kgf/cm2 for gas pressure. The scaffolds after particulate leaching yield porosity over 90% with pore size 200 ~ 500 μm, which satisfies what Health Medical chondrocytes need. Furthermore, the narrow version of the human-ear-shaped scaffold is also molded in this research. The mold is fabricated by rapid protyping and then by casting using PDMS. The transfer moldings of PLGA/NaCl using PDMS mold, after leaching, can yield ear-shaped scaffolds with pore size 200 ~ 500 μm. The disk scaffold is used to culture cartilage cells. The morphology and distribution of the chondrocytes have been examined by fluorescence confocal microscopy and SEM. The cell growth behaviors has been determined by MTT assay, collagen, and glycosaminoglycan (GAG) secretion. After culture for 1 day, 84% of cartilage cells attached on scaffold. This proves that the transfer molded PLGA/NaCl, after leaching, can yield scaffolds with interconnected pores for culturing cells. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/65239 |
全文授權: | 有償授權 |
顯示於系所單位: | 機械工程學系 |
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