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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 蔡偉博(Wei-Bor Tsai) | |
dc.contributor.author | Wei-Han Lin | en |
dc.contributor.author | 林威翰 | zh_TW |
dc.date.accessioned | 2021-06-16T22:56:51Z | - |
dc.date.available | 2014-08-21 | |
dc.date.copyright | 2012-08-21 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-08-10 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/64642 | - |
dc.description.abstract | 使用天然高分子材料製作電紡絲時,通常會具備易使細胞貼附、生物相容性高及具生物可分解性等優點,但較弱的機械性質則是其共有的缺點,而高度的水溶性也會限制天然高分子材料電紡絲在生醫材料方面的應用,現今的研究主要是利用各種物理或化學的交聯方法來改善這些缺點。但目前對於電紡絲的交聯方法研究仍存在著生物毒性不易去除或效率過低等問題。本研究使用聚丙烯酸接枝疊氮基(PAA-Az)與明膠(Gelatin) 混摻並進行電紡絲實驗,並在製程中以紫外光照射使其交聯,藉以建立一快速且低生物毒性之新型電紡絲交聯方法。並以纖維母細胞培養及混摻氫氧基磷灰石(HAp)後骨母細胞礦化結果來分析其細胞貼附和增生之表現及進一步修飾應用之可能性。最後於此明膠電紡絲中分別及共同混摻入氫氧基磷灰石、聚乙烯亞胺接枝RGD序列(PEI-RGD)及骨型態蛋白-2 (BMP-2),並在其上培養間葉幹細胞,以製作具骨組織工程發展潛力之生物支架。
結果發現,以聚丙烯酸接枝疊氮基交聯之明膠電紡絲機械性質及玻璃轉換溫度均較未交聯者為高;不具明顯細胞毒性,且纖維母細胞貼附和增生之表現也較以戊二醛交聯者為佳;混摻氫氧基磷灰石後骨母細胞之礦化表現有所上升(第14天約132%)。間葉幹細胞在含有氫氧基磷灰石或聚乙烯亞胺接枝RGD序列的電紡絲上也有鹼性磷酸酶活性(第7天約44%)和礦化表現提升(第14天約83%)或細胞貼附和增生之表現提升的相應表現。實驗結果說明此交聯方法應用於天然及水溶性高分子電紡絲製備與組織工程支架製作上具有相當的潛力。 | zh_TW |
dc.description.abstract | Electrospun fibers fabricated by natural polymer usually consisted of high cell adhesion ability with many beneficial characteristics, including their biocompatibility and biodegradability. However, weak mechanical property and high dissolution property are some of the drawbacks for natural polymer in limiting its application in electrospun fiber for biomedical usage. In addition, the biological toxicity was noted during the crosslinkage process of electrospun fibers remained a crucial problem in current research with either precipitation of toxic substances after crosslinkage or inefficient methods for removing them. Besides, the commonly used two-step crosslinking method (crosslinked after fiber fabrication) may cause un-even crosslinking of interior and exterior of the electrospun scaffolds. In this research, UV crosslinker poly(acrylic acid)-g-azide (PAA-Az) combined with an in situ crosslinking method was used for crosslinkage of gelatin to provide a low biological toxic route in fabricating electrospun fibers with high fibroblast proliferation. Hydroxyapatite (HAp) was also mixed in the solution for fabrication of the crosslinked fibers, resulting in high mineralization of osteoblasts. Finally, HAp, PEI-RGD, and BMP-2 were mixed in the solution for fabrication crosslinked electrospun gelatin fibers, resulting in high cell proliferation and mineralization of mesenchymal stem cells.
As result, gelatin electronspun fibers crosslinked by PAA-Az exhibited higher mechanical strength and glass transition temperature in compared to un-crosslinked gelatin. When comparing gelatin electrospun fibers crosslinked by glutaraldeyde, low cell toxicity was observed with enhancement in fibroblast adhesion for PAA-Az crosslinked gelatin fibers. In addition, when blending HAp in the solution, higher mineralization was also observed in osteoblasts on PAA-Az crosslinked gelatin fibers in compared to crosslinked gelatin fibers without HAp. Finally, PEI-RGD incorporation indeed enhance 3A6 cell attachment and proliferation. HAp incorporation increase the calcium deposited amount of 3A6 cell seeded on the electrospun gelatin fibers. The research presents potential of the in situ PAA-Az crosslinking method in the application of natural and high dissoluble polymeric electrospun fibers for tissue engineering. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T22:56:51Z (GMT). No. of bitstreams: 1 ntu-101-R99524051-1.pdf: 4341835 bytes, checksum: e539d0584b9ab49f76cad259920c75d4 (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | Abstract ii
List of Tables vii List of Figures viii Chapter 1 1 Introduction 1 1.1 Tissue Engineering 1 1.1.1 Overview of tissue engineering 1 1.1.2 Bone tissue engineering 2 1.1.3 Scaffolds for bone tissue engineering 3 1.1.4 Biofactors for bone tissue engineering 4 1.2 Electrospinning 6 1.2.1 Overview of electrospinning 6 1.2.2 Electrospinning parameters 8 1.2.3 Applications of electrospinning in tissue engineering 9 1.2.4 Gelatin as material of tissue engineered electrospun scaffolds 10 1.2.5 Enhance stability of eletrospun natural and water-soluble polymer fibers by crosslinking 11 1.2.6 Crosslinking of eletrospun natural and water-soluble polymer fibers by azide chemistry 13 1.3 Motive and aim 15 1.4 Research frame work 16 Chapter 2 17 Materials and Methods 17 2.1 Materials 17 2.1.1 Crosslinker synthesis and characterization 17 2.1.2 Electrospinning 17 2.1.3 Characterization of electrospun fiber 18 2.1.4 Cytotoxicity assay (3-(4,5-Dimethylthiazol-2-yl)-2,5- diphenyltetrazoliumbromide, MTT assay) 18 2.1.5 Mouse fibroblast-like cell line L929 culture 18 2.1.6 Cell number determination (Lactate dehydrogenase, LDH assay) 19 2.1.7 Synthesis of hydroxyapatite nanoparticles (HApNPs) 20 2.1.8 Osteoblast-like cell line MG63 culture 20 2.1.9 Alkaline phosphatase (ALP) activity 21 2.1.10 Mineralization culture 21 2.1.11 Quantification of calcium deposition 21 2.1.12 Synthesis of poly(ethylene imine) grafted RGD (PEI-g-RGD) 21 2.1.13 Human mesenchymal stem cell line (3A6) cell culture and osteogenic differentiation 22 2.2 Experimental instrument and materials 23 2.2.1 Experimental instrument 23 2.2.2 Experimental consumables 24 2.3 Solution formula 25 2.4 Methods 27 2.4.1 Synthesis of poly(acrylic acid)-g-Az (PAA-Az) UV-crosslinker 27 2.4.2 Fabrication of electrospun fiber meshes 28 2.4.3 Structure of uncrosslinked and crosslinked electrospun fiber meshes 31 2.4.4 Dissolvability of gelatin ESF meshes 32 2.4.5 Mechanical and physiochemical properties of gelatin electrospun fibers 32 2.4.6 Cytotoxicity analysis 33 2.4.7 Mouse fibroblast-like cell line L929 culturing 34 2.4.8 Cell density analysis 35 2.4.9 Synthesis of hydroxyapatite nanoparticles (HApNPs) 36 2.4.10 Hydroxyapatite nanoparticles (HApNPs) incorporated in gelatin electrospun fibers 37 2.4.11 Osteoblast-like cell line MG63 culturing 37 2.4.12 Alkaline phosphatase (ALP) activity 38 2.4.13 Calcium quantification 38 2.4.14 Synthesis of PEI-g-RGD 39 2.4.15 Statistic analysis 39 Chapter 3 41 Characterization and cell affinity analysis of crosslinked gelatin electrospun fibers 41 3.1 Synthesis of UV-crosslinker PAA-Az 41 3.2 Structure of uncrosslinked and crosslinked electrospun fiber meshes 42 3.3 Mechanical and physiochemical properties of gelatin electrospun fibers 44 3.4 Cytotoxic assay 46 3.5 L929 adhesion and proliferation on crosslinked gelatin fibers 46 3.6 Discussion 47 Chapter 4 61 Application of in-situ UV-crosslinked Gelatin Electrospun Fibers for Bone Tissue Engineering 61 4.1 Characterization of gelatin electrospun fibers with hydroxyapatite nanoparticles incorporation 61 4.2 MG63 proliferation, ALP expression and mineralization on HA incorporated crosslinked electrospun gelatin fibers 62 4.3 Morphology of gelatin electrospun fibers incorporated with HApNPs, PEI-RGD, and BMP-2 64 4.4 3A6 proliferation, ALP expression and mineralization on gelatin electrospun fibers incorporated with HApNPs, PEI-RGD, and BMP-2 65 4.5 Discussion 69 Chapter 5 87 Conclusion 87 Chapter 6 88 Future Works 88 Reference 89 Appendix 94 | |
dc.language.iso | zh-TW | |
dc.title | 紫外光臨場交聯明膠電紡絲應用於組織工程 | zh_TW |
dc.title | In situ UV-crosslinked Electrospun Gelatin Fibers for
Tissue Engineering | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 謝學真(Hsyue-Jen Hsieh),游佳欣(Jia-Shing Yu),賴瑞陽(Jui-Yang Lai) | |
dc.subject.keyword | 電紡絲,紫外光交聯,明膠,疊氮化物,組織工程, | zh_TW |
dc.subject.keyword | electrospinning,UV-crosslinking,gelatin,azide,tissue engineering, | en |
dc.relation.page | 96 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2012-08-10 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 化學工程學研究所 | zh_TW |
顯示於系所單位: | 化學工程學系 |
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