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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林思洸(Sze-Kwan Lin) | |
| dc.contributor.author | Ching-Lun Lin | en |
| dc.contributor.author | 林靖倫 | zh_TW |
| dc.date.accessioned | 2021-05-14T17:42:08Z | - |
| dc.date.available | 2018-09-24 | |
| dc.date.available | 2021-05-14T17:42:08Z | - |
| dc.date.copyright | 2015-09-24 | |
| dc.date.issued | 2015 | |
| dc.date.submitted | 2015-08-19 | |
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Brissault B, Kichler A, Guis C, Leborgne C, Danos O, and Cheradame H. Synthesis of Linear Polyethylenimine Derivatives for DNA Transfection. Bioconjug Chem. 2003;14:581-7. 45. Luo D, and Saltzman WM. Synthetic DNA Delivery Systems. Nat Biotechnol. 2000;18:33-7. 46. Boussif O, Lezoualc'h F, Zanta MA, Mergny MD, Scherman D, Demeneix B, and Behr JP. A Versatile Vector for Gene and Oligonucleotide Transfer into Cells in Culture and in Vivo: Polyethylenimine. Proc Natl Acad Sci U S A. 1995;92:7297-301. 47. Lungwitz U, Breunig M, Blunk T, and Gopferich A. Polyethylenimine-Based Non-Viral Gene Delivery Systems. Eur J Pharm Biopharm. 2005;60:247-66. 48. Sidi AA, Ohana P, Benjamin S, Shalev M, Ransom JH, Lamm D, Hochberg A, and Leibovitch I. Phase I/II Marker Lesion Study of Intravesical Bc-819 DNA Plasmid in H19 over Expressing Superficial Bladder Cancer Refractory to Bacillus Calmette-Guerin. J Urol. 2008;180:2379-83. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/4415 | - |
| dc.description.abstract | 引導組織再生術（guided tissue regeneration，GTR）已被廣泛應用於修復組織缺損的牙科手術治療中，但對於較大範圍的組織缺損，仍需缺損區有足夠的組織再生能力才可能癒合。過去利用在骨粉（bone graft）中加入生長因子（growth factor）來提升組織再生的能力，然而生長因子的使用仍面臨許多問題。但已知生長因子能夠促使幹細胞（stem cells）遷移（migration）、分化（differentiation），而利用吸引幹細胞以促進組織再生已成為目前主要的發展方向。基質細胞衍生因子-1α（stromal cell-derived factor-1α，SDF-1α）被認為是重要的幹細胞趨化素（chemokine），位在血管四周的間質幹細胞（mesenchymal stem cells，MSCs），能藉由產生基質細胞衍生因子-1α來維持造血幹細胞（hematopoietic stem cells，HSCs）的運送和成熟，與促進血管新生和骨再生都有相關。文獻也已證實其對軟組織和硬組織癒合都有正面影響，唯文獻多以蛋白質作為研究對象，蛋白質卻受限於作用效期短、安全性及成本昂貴等問題。因此本實驗以重新建構的SDF-1α質體（plasmid）取代SDF-1α蛋白質進行實驗，確認此質體確能釋放出SDF-1α蛋白質，並且和市售的SDF-1蛋白質一樣可以吸引幹細胞遷移。未來本實驗希望以聚乙烯亞胺（polyethylenimine，PEI）轉染（transfection）的方式，完成動物實驗，以期SDF-1α發揮未來在臨床上應用於再生技術上的優勢。 | zh_TW |
| dc.description.abstract | Guided tissue regeneration (GTR) has been widely used to repair tissue defects in oral surgery. The healing tendency, however, won’t occur without enough tissue regenerative potential in wider range of defects. In the past, growth factors were added into bone graft to enhance tissue regeneration, but the use of growth factors still faces many problems. It is known that the growth factor can promote stem cell migration and differentiation. Attracting stem cells in order to promote tissue regeneration has become the main direction of development. Stromal cell-derived factor-1α (SDF-1α) is considered an important stem cell chemokine. Mesenchymal stem cells (MSCs), which are located in perivascular position, can generate SDF-1α to maintain the delivery and maturity of hematopoietic stem cells (HSCs); furthermore, promoting angiogenesis and bone regeneration are related. Literature has also been confirmed that SDF-1α has a positive impact on soft tissue and hard tissue healing, though most of the studies using SDF-1α protein as the research subject. Protein was restricted to the role of the short lasting time, safety and costly; therefore, the construction of SDF-1α plasmid to replace SDF-1α protein was done in this experiments. The experiment confirmed this plasmid DNA can release SDF-1α protein, moreover, it can attract stem cell migration as commercial SDF-1α protein does. In future prospect, it is expect to be able to use polyethyleneimine (PEI) transfection method to complete animal study, in order that SDF-1α plays to its advantages in regenerative technique of future clinical applications. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-14T17:42:08Z (GMT). No. of bitstreams: 1 ntu-104-R01422020-1.pdf: 1142342 bytes, checksum: 13ad85eed681219ea9a37d12819d449b (MD5) Previous issue date: 2015 | en |
| dc.description.tableofcontents | 口試委員會審定書............................................................................................i
誌謝...............................................................................................................ii 中文摘要........................................................................................................iii 英文摘要........................................................................................................iv 目錄..............................................................................................................vi 第一章 導論....................................................................................................1 1.1 引導組織再生術與生長因子.........................................................................1 1.2 幹細胞與組織再生......................................................................................3 1.3 基質細胞衍生因子-1α................................................................................4 1.4 基因傳遞與活體影像追蹤............................................................................6 1.4.1 生物載體…............................................................................................6 1.4.2 非病毒技術............................................................................................7 1.4.3 非侵入式活體分子影像系統與螢光選擇.....................................................7 第二章 實驗目的 .............................................................................................9 第三章 材料與方法.........................................................................................10 3.1 試劑與抗體.............................................................................................10 3.2 實驗細胞株.............................................................................................10 3.3 基因重組.................................................................................................11 3.3.1 重組基因片段放大….............................................................................11 3.3.2 DNA接合反應.......................................................................................11 3.3.3 大腸桿菌的轉型....................................................................................11 3.3.4 確認pLenti-C-mCherry........................................................................12 3.3.5 重組pLenti-T2A-C-mCherry.................................................................12 3.3.6 重組pLenti-SDF-1-T2A-C-mCherry......................................................12 3.3.7 純化小量質體.......................................................................................13 3.3.8 純化大量質體.......................................................................................13 3.4 細胞轉染.................................................................................................14 3.5 不含細胞培養基的收集.............................................................................14 3.6 確認mCherry螢光...................................................................................14 3.7 細胞內蛋白質的萃取................................................................................14 3.8 SDF-1α的酶聯免疫吸附試驗.....................................................................16 3.9 西方點墨法.............................................................................................16 3.10 幹細胞遷移試驗.....................................................................................17 第四章 實驗結果............................................................................................18 4.1 螢光顯微鏡下mCherry的螢光表現.............................................................18 4.2 細胞內的mCherry蛋白質表現...................................................................18 4.3 不含細胞培養基的SDF-1α表現.................................................................18 4.4 SDF-1α蛋白質刺激VEGF表現..................................................................19 4.5 幹細胞遷移試驗.......................................................................................19 第五章 結論..................................................................................................20 第六章 討論..................................................................................................21 5.1 SDF-1蛋白質和質體傳遞之異同................................................................21 5.2 細胞轉染與聚乙烯亞胺的臨床應用.............................................................22 參考文獻......................................................................................................24 附錄.............................................................................................................32 | |
| dc.language.iso | zh-TW | |
| dc.title | 以基質細胞衍生因子-1 alpha在再生技術中促進骨生成與傷口癒合 | zh_TW |
| dc.title | Enhancement of bone formation and wound healing by stromal cell-derived factor-1 alpha in regenerative technique | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 103-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 郭生興,洪志遠 | |
| dc.subject.keyword | 基質細胞衍生因子-1,間質幹細胞,引導組織再生術,基因傳遞,慢病毒載體, | zh_TW |
| dc.subject.keyword | stromal cell-derived factor-1,mesenchymal stem cell,guided tissue regeneration,gene delivery,lentiviral vector, | en |
| dc.relation.page | 36 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2015-08-19 | |
| dc.contributor.author-college | 牙醫專業學院 | zh_TW |
| dc.contributor.author-dept | 臨床牙醫學研究所 | zh_TW |
| 顯示於系所單位： | 臨床牙醫學研究所 | |
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