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研究生:薛春連
研究生(外文):Chun-Lien Hsueh
論文名稱:生醫高分子支架於骨組織工程之研究與開發
論文名稱(外文):The Research and Development of PLGA scaffold for Tissue Engineering
指導教授:劉士榮劉士榮引用關係
學位類別:碩士
校院名稱:長庚大學
系所名稱:醫療機電工程研究所
學門:工程學門
學類:機械工程學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:中文
論文頁數:90
中文關鍵詞:支架 組織工程 PLGA
外文關鍵詞:scaffoldTissue engineeringbiodegradablePLGA
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臨床上骨組織工程在骨缺損病例上需要更好的填充物,而骨填充物需要優良的生物機械性質與良好的生物相容性,在傳統治療骨缺損方式上,不論是採用自體移植、異體移植上亦或是採用人造材料上,都會有些發炎感染或是生物相容性不佳的問題存在,受限於傳統療法的限制,發展骨組織工程可有效改善目前之瓶頸。在傳統的細胞培養方式是將細胞培養在2-D的環境下,讓細胞可生長並複製成大的聚落(colonies)來做應用。但是如此形成之聚落並不會有次序的組成組織或器官。而骨組織工程乃是應用3D的多孔性材料作為支架以培養幹細胞(Stem cell),來作為組織再生的模板,進而有次序的生成具功能性3D骨組織。
本研究之目的為開發一可促進骨細胞生長並具生物可降解性支架,其方法為利用本實驗室所開發之熱壓燒結法,來製備出PLGA多孔性支架,控制孔洞大小在80-125μm之範圍內,以利適合幹細胞成長。第一階段乃是利用物理檢驗方法,來探討本研究中所開發之熱壓成型法支架是否可讓細胞繁殖於上。第二階段則進行支架與細胞間的相互關係驗證,並探討不同孔徑支架對於細胞繁殖之影響。
Tissue engineering is an interdisciplinary and multidisciplinary field. It has showed great promise in generating living alternatives for harvested tissues and organs for transplantation and reconstructive surgery. Materials and fabrication technologies are critically important for tissue engineer in design temporary, artificial extracellular materials (scaffolds), which support three-dimensional tissue formation.

The report was to develop a novel solvent-free method for the manufacture of biodegradable scaffold. To manfacture a scaffold, PLGA(polylactide-polyglycolide) copolymers were pre-mixed with NaCl. The mixture was then injection compression molded to form a cylinder. Three particle sizes of NaCl (<88μm, 125-88μm, >125μm) were we used, while the PLGA to salt ratio is 1:9. Alkialine phosphatase(ALP) activity per cell were observed on cultured scaffold. The 88-125μm pore-size scaffold appears to be the most appropriate size for culturing after 7 days. The fluorometric quantification of DNA suggested that the 88-125μm size scaffold is the superior one for cell culture at 7 day.


Keywords : scaffold; Tissue engineering; biodegradable;PLGA
摘要
目錄
第一章 序論……………………………………………………….. …..1
1.1 組織工程簡介………………………………………………3
1.2 骨組織缺陷修復簡介…………………………………………...7
1.3 生醫材料簡介………………………………………………….11
1.4 幹細胞簡介...…………………………………………………15
1.5 PLGA簡介……………………………………………………21
1.6 三維立體支架製備簡介……………………………………....26
1.7 研究動機與目的………………………………………………33
1.8 論文架構……………………………………………………….35

第二章 文獻回顧……………………………………………..36
2. 1高分子材料文獻回顧……………………………………36
2.2支架製備方式文獻回顧……………………………………….38
2.3文獻總結……………………………………………………….40

第三章 實驗材料與方法…………………………………………. 42
3.1實驗材料………………………………… 43
3.2實驗過程………………………………... 47
3.3PLGA-NaCl支架測試……………………………. 49
3.4幹細胞培養定性……………………………………………….56
3.5鹼性磷酸活性測試…………………………………………….58
3.7 DNA content……………………………………………………….59

第四章 研究結果……………………………………………. 61
4.1製程可行性驗證………………………................................. 61
4.2多孔性支架觀察……………………………………... 62
4.3細胞-支架定量分析……………………………………….…...79

第五章 結論..………………………………………………………...86
5.1 實驗結論………………………………………………………86
5.2未來研究目標………………………………………………….87

第六章 參考文獻…………………………………………….………....89
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[14] Lu-LC, Peter-SJ, Lyman-MD, Lai-HL, Leite-SM, Tamada-JA,
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[15] 王詩涵, “以超臨界二氧化碳反溶劑法製備PLGA微粒”
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[16] 吳侑俊,”骨的組織工程研究-利用聚乳酸-聚乙醇酸共聚物當作支架”,成功大學生物科技研究所碩士論文,92

[17]Susan L. Ishaug,1, Genevieve M. Crane, Michael J. Miller,Alan W. Yasko,Michael J. Yaszemski,, andAntonios G. Mikos, ”Bone formation by three-dimensional stromal osteoblastculture in biodegradable polymer scaffolds”, Journal of Biomedical Materials Research, Vol. 36, 17–28 ,1997

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[20]Kota Uematsu, Koji Hattori, Yoshiyuki Ishimoto, Jun Yamauchi, Takashi Habata,Yoshinori Takakura, Hajime Ohgushi, Takeshi Fukuchi,Masao Sato.” Cartilage regeneration using mesenchymal stem cells and athree-dimensional poly-lactic-glycolic acid (PLGA) scaffold”. Biomaterials26 ,4273–4279,2005
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