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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 林?輝 | |
dc.contributor.author | Chi-Sain Fu | en |
dc.contributor.author | 傅紀珊 | zh_TW |
dc.date.accessioned | 2021-06-13T01:31:12Z | - |
dc.date.available | 2007-07-23 | |
dc.date.copyright | 2007-07-23 | |
dc.date.issued | 2007 | |
dc.date.submitted | 2007-07-17 | |
dc.identifier.citation | 第六章 參考文獻
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Differentiation of human embryonic stem cells into embryoid bodies compromising the three embryonic germ layers. Mol Med. 2000; 2: 88-95. 9. S. Gronthos, J. Brahim J, W. Li, L.W. Fisher, N. Cherman, A. Boyde, P. DenBesten, P. Gehorn Robey and S. Shi. Stem cell properties of human dental pulp stem cells. J Dent Res 2002; 8: 531-535. 10. S. Gronthos, M. Mankani, J. Brahim, P. Gehorn ans S. Shi. Postnatal human dental pulp stem cells ( DPSCs ) in vitro and in vivo. Proc. Natl. Acad. Sci 2000; 97: 13625-13630. 11. M.S. Miura, S. Gronthos, M. Zhao, B. Lu, L.W. Fisher, P.G. Robey and S. Shi. SHED: stem cells from human exfoliated deciduous teeth. Proc. Natl. Acad. Sci 2003; 100︰ 5807-5812. 12. A. Ohazama, S.A.C. Modino, I. Miletich and P.T Sharpe. Stem-cell-based tissue engineering of murine teeth. J Dent Res 2004; 7: 518-522. 13. M.T. Dualibi, S.E. Duailibi, C.S. Young, J.D. Bartlett, J.P. Vacanti and P.C. Yelick. Bioengineered Teeth from Cultures Rat Tooth Bud Cells. J Dent Res 2004﹔83﹕523-528. 14. R.L. Reis and J.S. Roman. Biodegradable Systems in Tissue Engineering and Regenerative Medicine. CRC Press 2005. 15. R.P. Lanza, R. Langer and J. Vacanti. Principles of Tissue Engineering 2nd.2000﹔xxxvi-xli. 16. P. Nieminen, M. Pekkanen, T. Aberg and I. Thesleffe. A graphical WWW-database on gene expression in tooth. Eur J Oral Sci 1998﹔106 Suppl 1﹕7-11. 17. J. Jernvall, I. Thesleff. Reiterative signaling and patterning during mammalian tooth morphogenesis. Mech Dev 2000﹔92﹕19-29. 18. M. Nakashima, A.H. Reddi. The application of bone morphogenetic proteins to dental tissue engineering. Nat Biotechnol 2003﹔21﹕1025-1032. 19. A. Neubuser , R. Balling, H. Peters and G.R. Martin. Antagonistic interactions between FGF and BMP signaling pathway﹕a mechanism for positioning the sites of tooth formation. Cell 1997﹔90﹕247-255. 20. P. Ketturn and I. Thesleff. Expression and function of FGFs-4,-8 and –9 suggest functional redundancy and repetitive use as epithelial signals during tooth morphogenesis. Dev Dyn 1998﹔211﹕256-268. 21. Michel Goldberg and Anthony J. Smith. Cell and extracellylar matrices of dentin and pulp﹕a biological basis for repair and tissue engineering. Crit Rev Oral Biol Med 2004﹔15( 1 )﹕13-27. 22. M. MacDougall, D. Simmins, X. Luan, J. Nydegger, J. Feng and T.T. Gu. Dentin phosphoprotein and dentin sialoprotein are cleaved products expressed from a single transcript coded by a gene on human chromosome 4. J Biol Chem 1997﹔272﹕835-842. 23. C. Qin, J.C. Brunn, E. Cadena, A. Ridall, H. Tsujigiwa and H. Nagatsuka. The expression of dentin sialophosphoprotein gene in bone. J Dent Res 2002﹔81﹕392-394. 24. A. Gerorge, B. Sabsay, P.A.L. Simonian and A. Veis. Characterization of a noel dentin matrix acidic phosphoprotein. J Biol Chem 1993﹔268﹕12642-12630. 25. 周正鴻。明膠/透明質酸/軟骨素共聚物做為關節軟骨組織工程海綿狀支架之研究。國立臺灣大學醫學工程研究所碩士論文。2003。 26. E Ruoslanti. RGD and other recognition sequences for integrins, Annual Review of Cell & Developmental Biology. 1996﹔12﹕697-715. 27. S Maniwa, M Ochi, T. Motomura, T. Nishikori, J. Chen and H. Naora. Effects of hyaluronic acid and basic fibroblast growth factor on motility of chondrocytes and synovial cells in culture. Acta Orthopaedica Scandinavica. 2001﹔72( 3 )﹕299-303. 28. I.M. Barbash, P. Chouraqui, J. Baron, M.S. Feinberg, S. Etzion, A. Tessone, L. Miller, E. Guetta, D. Zipori, L.H. Kedes, R.A. Kloner and J.L. Leor. Systemic delivery of bone marrow-derived mesenchymal stem cells to the infarcted myocardium: feasibility, cell migration, and body distribution. Circulation 2003; 7: 863-868. 29. S.N. Park, J.C. Park, H.O. Kim, M.J. Song and H. Suh. Characterization of porous collagen/hyaluronic acid sxaffold modified by 1-ethyl-2-( 3-dimethylaminopropyl ) carbodiimide cross-linking. Biomaterials 2002﹔23( 4 )﹕1205-1212. 30. A. Aala, R.P. Lanza. Methods of tissue engineering. Academic Press 2002﹔99-108. 31. D.M. Huang, Y. Hung, B.S. Ko, S.H. Hsu, W.H. Chen, C.L. Chien, C.P. Tsai, C.T. Kuo, J.C. Kang, C.S. Yang, C.Y. Mou and Y.C. Che. Highly efficient cellular labeling of mesoporous nanoparticles in human mesenchymal stem cells: implication for stme cell tranking. The FASEB journal.2005. 32. Pierdomenico L. Multipotent mesenchymal stem cells with immunosuppressive activity can be easily isolated from dental pulp. ransplantation 2005; 80: 836-842. 33. Roche. Cell proliferation reagent WST-1 protocal, 2005. 34. MK Campbell, John Vondeling. Biochemistry 1999﹔466-474. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30023 | - |
dc.description.abstract | 牙齒在攝取營養、外觀及與人溝通方面,都是不可或缺的。齲齒、牙髓炎、牙周疾病及外傷等因素,將導致牙齒缺失,進而影響正常咀嚼、發音等生理功能,並造成外型上的困擾。若是能發展出生物性的替代牙齒便能取代缺損的牙齒,將是臨床治療的一大突破;而幹細胞可遷移至器官受損處進行修復,並具有可自我更新及分化的潛力,若能運用幹細胞組織工程技術來達到此目標,將是一大進展。
為了評估這個目標,我們取出大鼠臼齒的牙胚細胞與大鼠骨髓間葉幹細胞分離成單細胞方式共養後,評估有無共養之細胞活性及對牙生成有幫助的相關基因表現之比較。共養之細胞與牙胚細胞分別接種在具生物分解性的雙層支架中並植入免疫缺失鼠背部皮下4、8、12週後,以X光放射線攝影、組織化學染色及免疫組織化學染色進行檢測。 實驗結果顯示,大鼠骨髓間葉幹細胞經鑑定後確定其幹細胞之特性,而經共養之後的大鼠骨髓間葉幹細胞與大鼠牙胚細胞在細胞活性及基因表現量上都有提升。第12週共養之細胞與牙胚細胞由X光放射線攝影圖中看到硬組織的產生、組織化學染色中有表現鈣鹽,推測已有礦化的產生。在第4、8、12週時,共養之細胞與牙胚細胞在免疫組織化學染色都有表現Dentin Matrix Protein-1 ( DMP-1)蛋白,確定已有類牙本質的產生,而從染色結果得知,共養之細胞在DMP-1蛋白表現量遠高於牙胚細胞。可確定的是共養之細胞與牙胚細胞在具生物分解性的雙層支架中,大鼠骨髓間葉幹細胞與牙胚細胞可以進行重組而形成礦化與類牙本質之結構。 | zh_TW |
dc.description.abstract | The human dentition is indispensable for nutrition, communication and physiology. Caries, pulpitis, apical periodontitis and trauma may lead to loss of tooth, causing the problem of pronunciation, mastication, and appearance difficulties. A biological tooth substitute that could replace lost teeth would provide a vital alternative to currently available clinical treatment. Stem cells are multipotent cells which are capable of self-renewing and differentiating into multi- cell lineages. In this study, tissue engineering approach will be addressed to evaluate this possibility to reconstruct tooth.
To achieve this goal, we separately dissociated Wistar rat Tooth Bud Cells and Mesenchymal Stem Cells ( MSC ). Then two kinds of cells were seeded either in monoculture ( Tooth Bud Cells only ) or co-culture ( MSC + Tooth Bud Cells ) condition to evaluate cell viability and gene expression in vitro. Co-culture Cells and Tooth Bud Cells were then respectively seeded onto biodegradable bilayered scaffold and implanted in subcutaneous layers in SCID mice. After implantation for 12 weeks, the results were studied by X ray, histology and immunohistochemistry. From the results, the cell phenotype of rat msenchymal stem cells was identified by flow cytometry. Cells viability and gene expression were elevated in co-culture group. At 12 weeks, the mineralization in Co-culture Cells group and Tooth Bud Cells group was visualized in X ray, Von Kossa and Alizarin Red S. At 4, 8, 12 weeks, the dentin matrix protein-1 ( DMP-1) were deposited and stained in Co-culture Cells group and Tooth Bud Cells group by immunohistochemistry, representing dentin-like structure formation. In addition, Co-culture Cells group expressed stronger than Tooth Bud Cells group. It seems like that rat Mesenchymal Stem Cells and Tooth Bud Cells could reorganize themselves on biodegradable bilayered scaffold into arrangements that favor formation of mineralization / dentin-like tissue. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T01:31:12Z (GMT). No. of bitstreams: 1 ntu-96-R94548001-1.pdf: 2160731 bytes, checksum: c43e63bf409470de81780a0cb9f676ff (MD5) Previous issue date: 2007 | 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 圖目錄……………………………………………………………...IX 表目錄……………………………………………………………...XII 第一章 簡介…………………………………………………….1 1-1 前言………………………………………………………1 1-2 牙齒的形成………………………………………………1 1-3 牙齒的基本構造…………………………………………2 1-4 幹細胞……………………………………………………5 1-5 文獻回顧…………………………………………………8 1-6 研究目的…………………………………………………10 第二章 基本理論…………………………………………………11 2-1 組織工程…………………………………………………………11 2-2 牙齒的基因調控…………………………………………………12 2-3 牙本質的組成……………………………………………………13 2-4 支架材料的選擇…………………………………………………15 第三章 實驗方法…………………………………………………16 3-1 實驗儀器…………………………………………………………16 3-2 實驗藥品…………………………………….………………….17 3-3 實驗流程與方法…………………………………………………18 3-4 細胞培養…………………………………………………………21 3-4-1 大鼠骨髓間葉幹細胞的分離與培養方法………………21 3-4-2 大鼠牙胚細胞的分離與培養方法………………………21 3-4-3 大鼠骨髓間葉幹細胞與牙胚細胞共養…………………22 3-5 支架製作…………………………………………………………23 3-6 細胞的接種………………………………………………………26 3-7 動物實驗…………………………………………………………26 3-8 分析方法…………………………………………………………27 3-8-1 流氏細胞儀鑑定大鼠間葉幹細胞………………………27 3-8-2 Total DNA與WST-1細胞增生及活性測試…………….29 3-8-2-1 Total DNA細胞增生…………………………….29 3-8-2-2 WST-1細胞活性測試…………………………...29 3-8-3 即時反轉錄聚合酶鍊鎖反應 ( real-time RT-PCR )…….31 3-8-4 材料分析…………………………………………………31 3-8-4-1 掃描式電子顯微鏡分析………………………...31 3-9 實驗動物皮下材料X光放射線攝影…………………………...32 3-10 組織化學分析…………………………………………………..32 3-10-1 石蠟包埋組織切片製作………………………………..32 3-10-1-1 自動儀器處理原則及步驟…………………….32 3-10-1-2 石蠟塊之包埋匣 ( cassette ) 包埋……………32 3-10-1-3 組織切片機…………………………………….33 3-10-2 組織切片脫蠟…………………………………………..34 3-10-3 蘇木紫-伊紅 ( Hematoxylin-eosin, H&E ) 染色………34 3-10-4 Von Kossa染色………………………………………….35 3-10-5茜素紅S ( Alizarin Red S )染色………………………...35 3-10-6 免疫組織化學分析……………………………………...36 第四章 結果與討論……………………………………………38 4-1 幹細胞之純化與培養……………………………………38 4-1-1 大鼠骨髓間葉幹細胞……………………………………38 4-1-2 大鼠牙胚細胞……………………………………………39 4-2 流式細胞儀鑑定大鼠骨髓間葉幹細胞…………………………40 4-3 Total DNA與WST-1細胞增生及活性測試…………………….41 4-4 即時反轉錄聚合酶鍊鎖反應 ( real-time RT-PCR )……….44 4-5 支架材料之分析…………………………………………………47 4-5-1 支架之掃描式電子顯微鏡分析…………………………47 4-6體內 ( in vivo ) 皮下培養…………………………….48 4-6-1 實驗動物皮下材料X光放射線攝影………………49 4-6-1-1 第4、8週之實體影像……………………………49 4-6-1-2 第12週實體影像及X光放射線攝影……..49 4-6-2 組織化學染色………………………………………52 4-6-3 免疫組織化學分析…………………………………59 第五章 結論……………………………………………………..62 第六章 參考文獻………………………………………………..63 | |
dc.language.iso | zh-TW | |
dc.title | 以組織工程技術於牙本質再生之應用 | zh_TW |
dc.title | The Application of Tissue Engineering on Dentin Regeneration | en |
dc.type | Thesis | |
dc.date.schoolyear | 95-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 林俊彬 | |
dc.contributor.oralexamcommittee | 傅紀珊 | |
dc.subject.keyword | 組織工程,骨髓間葉幹細胞,牙胚細胞,雙層支架,礦化,類牙本質, | zh_TW |
dc.subject.keyword | tissue engineering,Mesenchymal Stem Cells,Tooth Bud Cells,bilayered scaffold,mineralization,dentin-like, | en |
dc.relation.page | 80 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2007-07-17 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
顯示於系所單位: | 醫學工程學研究所 |
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