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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 鄭景暉 | |
dc.contributor.author | Shih-Han Yu | en |
dc.contributor.author | 尤詩涵 | zh_TW |
dc.date.accessioned | 2021-06-16T16:04:46Z | - |
dc.date.available | 2013-09-24 | |
dc.date.copyright | 2013-09-24 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-06-25 | |
dc.identifier.citation | References
Adams, C.L., and Nelson, W.J. (1998). Cytomechanics of cadherin-mediated cell-cell adhesion. Curr Opin Cell Biol 10, 572-577. Arnott, J.A., Nuglozeh, E., Rico, M.C., Arango-Hisijara, I., Odgren, P.R., Safadi, F.F., and Popoff, S.N. (2007). Connective tissue growth factor (CTGF/CCN2) is a downstream mediator for TGF-beta1-induced extracellular matrix production in osteoblasts. J Cell Physiol 210, 843-852. Assoian, R.K., Komoriya, A., Meyers, C.A., Miller, D.M., and Sporn, M.B. (1983). Transforming growth factor-beta in human platelets. Identification of a major storage site, purification, and characterization. J Biol Chem 258, 7155-7160. Bonewald, L.F., and Mundy, G.R. (1990). Role of transforming growth factor-beta in bone remodeling. Clin Orthop Relat Res, 261-276. Butler, W.T., and Ritchie, H. (1995). The nature and functional significance of dentin extracellular matrix proteins. Int J Dev Biol 39, 169-179. Campbell, I.D., and Humphries, M.J. (2011). Integrin structure, activation, and interactions. Cold Spring Harb Perspect Biol 3. Cheifetz, S., Weatherbee, J.A., Tsang, M.L., Anderson, J.K., Mole, J.E., Lucas, R., and Massague, J. (1987). The transforming growth factor-beta system, a complex pattern of cross-reactive ligands and receptors. Cell 48, 409-415. Derda, R., Li, L., Orner, B.P., Lewis, R.L., Thomson, J.A., and Kiessling, L.L. (2007). Defined substrates for human embryonic stem cell growth identified from surface arrays. ACS Chem Biol 2, 347-355. Engler, A.J., Sen, S., Sweeney, H.L., and Discher, D.E. (2006). Matrix elasticity directs stem cell lineage specification. Cell 126, 677-689. Escobedo-Lucea, C., Ayuso-Sacido, A., Xiong, C., Prado-Lopez, S., del Pino, M.S., Melguizo, D., Bellver-Estelles, C., Gonzalez-Granero, S., Valero, M.L., Moreno, R., et al. (2012). Development of a human extracellular matrix for applications related with stem cells and tissue engineering. Stem Cell Rev 8, 170-183. George, A., and Ravindran, S. (2010). Protein Templates in Hard Tissue Engineering. Nano Today 5, 254-266. Graycar, J.L., Miller, D.A., Arrick, B.A., Lyons, R.M., Moses, H.L., and Derynck, R. (1989). Human transforming growth factor-beta 3: recombinant expression, purification, and biological activities in comparison with transforming growth factors-beta 1 and -beta 2. Mol Endocrinol 3, 1977-1986. Grotendorst, G.R. (1997). Connective tissue growth factor: a mediator of TGF-beta action on fibroblasts. Cytokine Growth Factor Rev 8, 171-179. Guo, W., He, Y., Zhang, X., Lu, W., Wang, C., Yu, H., Liu, Y., Li, Y., Zhou, Y., Zhou, J., et al. (2009). The use of dentin matrix scaffold and dental follicle cells for dentin regeneration. Biomaterials 30, 6708-6723. Hao, J., Zou, B., Narayanan, K., and George, A. (2004). Differential expression patterns of the dentin matrix proteins during mineralized tissue formation. Bone 34, 921-932. Heino, J., Ignotz, R.A., Hemler, M.E., Crouse, C., and Massague, J. (1989). Regulation of cell adhesion receptors by transforming growth factor-beta. Concomitant regulation of integrins that share a common beta 1 subunit. J Biol Chem 264, 380-388. Huang, G.T., Sonoyama, W., Chen, J., and Park, S.H. (2006). In vitro characterization of human dental pulp cells: various isolation methods and culturing environments. Cell Tissue Res 324, 225-236. Huang, G.T., Sonoyama, W., Liu, Y., Liu, H., Wang, S., and Shi, S. (2008). The hidden treasure in apical papilla: the potential role in pulp/dentin regeneration and bioroot engineering. J Endod 34, 645-651. Huang, G.T., Yamaza, T., Shea, L.D., Djouad, F., Kuhn, N.Z., Tuan, R.S., and Shi, S. (2010). Stem/progenitor cell-mediated de novo regeneration of dental pulp with newly deposited continuous layer of dentin in an in vivo model. Tissue Eng Part A 16, 605-615. Hynes, R.O. (2002). Integrins: bidirectional, allosteric signaling machines. Cell 110, 673-687. Iohara, K., Zheng, L., Ito, M., Ishizaka, R., Nakamura, H., Into, T., Matsushita, K., and Nakashima, M. (2009). Regeneration of dental pulp after pulpotomy by transplantation of CD31(-)/CD146(-) side population cells from a canine tooth. Regen Med 4, 377-385. I-Hua Wu. (2011). Effect of TGF-β1 on the growth and differentiation of human apical papilla cells. Janssens, K., ten Dijke, P., Janssens, S., and Van Hul, W. (2005). Transforming growth factor-beta1 to the bone. Endocr Rev 26, 743-774. Lee-Thedieck, C., Rauch, N., Fiammengo, R., Klein, G., and Spatz, J.P. (2012). Impact of substrate elasticity on human hematopoietic stem and progenitor cell adhesion and motility. J Cell Sci 125, 3765-3775. Leivonen, S.K., Hakkinen, L., Liu, D., and Kahari, V.M. (2005). Smad3 and extracellular signal-regulated kinase 1/2 coordinately mediate transforming growth factor-beta-induced expression of connective tissue growth factor in human fibroblasts. J Invest Dermatol 124, 1162-1169. Li, R., Guo, W., Yang, B., Guo, L., Sheng, L., Chen, G., Li, Y., Zou, Q., Xie, D., An, X., et al. (2011). Human treated dentin matrix as a natural scaffold for complete human dentin tissue regeneration. Biomaterials 32, 4525-4538. Linde, A. (1989). Dentin matrix proteins: composition and possible functions in calcification. Anat Rec 224, 154-166. Loftis, M.J., Sexton, D., and Carver, W. (2003). Effects of collagen density on cardiac fibroblast behavior and gene expression. J Cell Physiol 196, 504-511. Lyons, R.M., and Moses, H.L. (1990). Transforming growth factors and the regulation of cell proliferation. Eur J Biochem 187, 467-473. Massague, J. (1990). The transforming growth factor-beta family. Annu Rev Cell Biol 6, 597-641. Melikova, S., Dylla, S.J., and Verfaillie, C.M. (2004). Phosphatidylinositol-3-kinase activation mediates proline-rich tyrosine kinase 2 phosphorylation and recruitment to beta1-integrins in human CD34+ cells. Exp Hematol 32, 1051-1056. Mih, J.D., Marinkovic, A., Liu, F., Sharif, A.S., and Tschumperlin, D.J. (2012). Matrix stiffness reverses the effect of actomyosin tension on cell proliferation. J Cell Sci 125, 5974-5983. Mio, T., Adachi, Y., Romberger, D.J., Ertl, R.F., and Rennard, S.I. (1996). Regulation of fibroblast proliferation in three-dimensional collagen gel matrix. In Vitro Cell Dev Biol Anim 32, 427-433. Nakashima, M. (1992). The effects of growth factors on DNA synthesis, proteoglycan synthesis and alkaline phosphatase activity in bovine dental pulp cells. Arch Oral Biol 37, 231-236. Nakashima, M., and Akamine, A. (2005). The application of tissue engineering to regeneration of pulp and dentin in endodontics. J Endod 31, 711-718. Nakashima, M., Nagasawa, H., Yamada, Y., and Reddi, A.H. (1994). Regulatory role of transforming growth factor-beta, bone morphogenetic protein-2, and protein-4 on gene expression of extracellular matrix proteins and differentiation of dental pulp cells. Dev Biol 162, 18-28. Narayanan, K. (2001). Differentiation of embryonic mesenchymal cells to odontoblast-like cells by overexpression of dentin matrix protein 1. Proceedings of the National Academy of Sciences 98, 4516-4521. Nelson, C.M., and Chen, C.S. (2003). VE-cadherin simultaneously stimulates and inhibits cell proliferation by altering cytoskeletal structure and tension. J Cell Sci 116, 3571-3581. Park, J.S., Chu, J.S., Tsou, A.D., Diop, R., Tang, Z., Wang, A., and Li, S. (2011). The effect of matrix stiffness on the differentiation of mesenchymal stem cells in response to TGF-beta. Biomaterials 32, 3921-3930. Prescott, R.S., Alsanea, R., Fayad, M.I., Johnson, B.R., Wenckus, C.S., Hao, J., John, A.S., and George, A. (2008). In vivo generation of dental pulp-like tissue by using dental pulp stem cells, a collagen scaffold, and dentin matrix protein 1 after subcutaneous transplantation in mice. J Endod 34, 421-426. Prosecka, E., Rampichova, M., Vojtova, L., Tvrdik, D., Melcakova, S., Juhasova, J., Plencner, M., Jakubova, R., Jancar, J., Necas, A., et al. (2011). Optimized conditions for mesenchymal stem cells to differentiate into osteoblasts on a collagen/hydroxyapatite matrix. J Biomed Mater Res A 99, 307-315. Puceat, M. (2007). TGFβ in the differentiation of embryonic stem cells. Cardiovascular Research 74, 256-261. Ranchalis, J.E., Gentry, L., Ogawa, Y., Seyedin, S.M., McPherson, J., Purchio, A., and Twardzik, D.R. (1987). Bone-derived and recombinant transforming growth factor beta's are potent inhibitors of tumor cell growth. Biochem Biophys Res Commun 148, 783-789. Shiba, H., Fujita, T., Doi, N., Nakamura, S., Nakanishi, K., Takemoto, T., Hino, T., Noshiro, M., Kawamoto, T., Kurihara, H., et al. (1998). Differential effects of various growth factors and cytokines on the syntheses of DNA, type I collagen, laminin, fibronectin, osteonectin/secreted protein, acidic and rich in cysteine (SPARC), and alkaline phosphatase by human pulp cells in culture. J Cell Physiol 174, 194-205. Sonoyama, W., Liu, Y., Fang, D., Yamaza, T., Seo, B.M., Zhang, C., Liu, H., Gronthos, S., Wang, C.Y., Wang, S., et al. (2006). Mesenchymal stem cell-mediated functional tooth regeneration in swine. PLoS One 1, e79. Sonoyama, W., Liu, Y., Yamaza, T., Tuan, R.S., Wang, S., Shi, S., and Huang, G.T. (2008). Characterization of the apical papilla and its residing stem cells from human immature permanent teeth: a pilot study. J Endod 34, 166-171. Tarone, G., Hirsch, E., Brancaccio, M., De Acetis, M., Barberis, L., Balzac, F., Retta, S.F., Botta, C., Altruda, F., and Silengo, L. (2000). Integrin function and regulation in development. Int J Dev Biol 44, 725-731. Togo, S., Sato, T., Sugiura, H., Wang, X., Basma, H., Nelson, A., Liu, X., Bargar, T.W., Sharp, J.G., and Rennard, S.I. (2011). Differentiation of embryonic stem cells into fibroblast-like cells in three-dimensional type I collagen gel cultures. In Vitro Cell Dev Biol Anim 47, 114-124. Tsuchiya, S., Honda, M.J., Shinohara, Y., Saito, M., and Ueda, M. (2007). Collagen type I matrix affects molecular and cellular behavior of purified porcine dental follicle cells. Cell and Tissue Research 331, 447-459. Tziafas, D., Alvanou, A., Papadimitriou, S., Gasic, J., and Komnenou, A. (1998). Effects of recombinant basic fibroblast growth factor, insulin-like growth factor-II and transforming growth factor-beta 1 on dog dental pulp cells in vivo. Arch Oral Biol 43, 431-444. Viale-Bouroncle, S., Vollner, F., Mohl, C., Kupper, K., Brockhoff, G., Reichert, T.E., Schmalz, G., and Morsczeck, C. (2011). Soft matrix supports osteogenic differentiation of human dental follicle cells. Biochem Biophys Res Commun 410, 587-592. Yang, B., Chen, G., Li, J., Zou, Q., Xie, D., Chen, Y., Wang, H., Zheng, X., Long, J., Tang, W., et al. (2012). Tooth root regeneration using dental follicle cell sheets in combination with a dentin matrix - based scaffold. Biomaterials 33, 2449-2461. Yeung, T., Georges, P.C., Flanagan, L.A., Marg, B., Ortiz, M., Funaki, M., Zahir, N., Ming, W., Weaver, V., and Janmey, P.A. (2005). Effects of substrate stiffness on cell morphology, cytoskeletal structure, and adhesion. Cell Motil Cytoskeleton 60, 24-34. Zhang, W., Frank Walboomers, X., van Kuppevelt, T.H., Daamen, W.F., Bian, Z., and Jansen, J.A. (2006). The performance of human dental pulp stem cells on different three-dimensional scaffold materials. Biomaterials 27, 5658-5668. Zhao, S., Sloan, A.J., Murray, P.E., Lumley, P.J., and Smith, A.J. (2000). Ultrastructural localisation of TGF-beta exposure in dentine by chemical treatment. Histochem J 32, 489-494. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62575 | - |
dc.description.abstract | 實驗目的:牙齒再生組織工程中,細胞生長因子和細胞支架對細胞生長、分化、胞外間質形成以及牙本質生成扮演著重要角色。本實驗之目的在於探討β1型轉型生長因子(Transforming growth factor-β1, TGF-β1)和膠原蛋白(Collagen)對牙根尖細胞生長與分化所造成的影響。我們假設膠原蛋白可以影響根尖細胞的貼附、活性、分化、鹼性磷酸酶的表現。此外,在體外的環境將根尖細胞培養在細胞支架中,並加入轉型生長因子可以成功地刺激組織再生。
實驗方法:第一部分使用不同濃度的膠原蛋白培養人類牙根尖細胞 (Stem cells from apical papilla, SCAP),檢測細胞的貼附能力並觀察細胞形態的改變,同時也以MTT測量細胞的活性並利用鹼性磷酸酶染色 (Alkaline phosphatase staining)觀察細胞分化的情形。第二部分使用不同濃度的TGF-β1和人類牙根尖細胞培養,以反轉錄聚合酶反應(RT-PCR)觀察結締組織生長因子(CTGF)和α1型組合蛋白(α1-integrin)的表現。最後將人類牙根尖細胞培養在兩種細胞支架(collagen/dentin scaffold)中並加入TGF-β1的刺激,以組織切片方式觀察細胞型態以及新生成的組織結構。 實驗結果:人類牙根尖細胞在覆有膠原蛋白的表面有助於細胞貼附的效果;在低濃度膠原蛋白表面生長會促進細胞的生長,而在高濃度膠原蛋白表面細胞活性會降低;鹼性磷酸酶染色也發現可看到根尖細胞在高濃度膠原蛋白的環境中鹼性磷酸酶的活性會略微降低。人類牙根尖細胞經過TGF-β1的刺激之後,CTGF和α1-integrin的表現量會隨著TGF-β1濃度升高而增加。將細胞培養在膠原蛋白細胞支架內再加入TGF-β1的刺激後初步結果可觀察到新生成的細胞間質堆積物;進一步將細胞放入牙本質細胞支架(dentin scaffold)中也可看到類似牙本質(dentin-like)的組織產生。 結論:TGF-β1和膠原蛋白對人類牙根尖細胞的附著、生長和分化都有調控的作用,在不同的條件配合下也可能誘導細胞產生不同的特性和分化的方向。在牙齒組織再生工程中,膠原蛋白和牙本質都是方便操作且具有潛力的細胞支架,若能合併適當的細胞和生長因子,未來可進一步應用在動物模型上甚至是臨床治療上,針對牙髓壞死的牙齒誘導牙本質和牙髓的再生。 | zh_TW |
dc.description.abstract | Aim: In tissue regeneration, growth factors and scaffolds play important roles in cell proliferation, differentiation, extracellular matrix production and dentin formation. The purpose of this study is to investigate the effects of TGF-β1 and collagen on behaviors of stem cells from apical papilla (SCAP). We hypothesize that collagen can affect cell attachment, viability, and alkaline phosphatase (ALP) expression in SCAP. Besides, culturing SCAP in scaffold with combination of TGF-β1 can regenerate tissues successfully in vitro.
Materials and Methods: SCAP were cultured on the surface coated with different densities of collagen. Then, SCAP were evaluated for cell attachment ability, cell viability by MTT test and cell differentiation by ALP activity. Besides, SCAP were treated with TGF-β1 with different concentrations. CTGF and α1-integrin expression were determined by reverse-transcription polymerase chain reaction(RT-PCR). Finally, we cultured SCAP in collagen scaffolds and dentin disc scaffolds with combination of TGF-β1 to observe the morphological changes of cells and tissue regeneration under histological analysis. Results: SCAP cultured on surfaces coated with collagen showed better attachment ability. SCAP viability elevated when cultured on the surface coated with lower densities of collagen. However, decreased cell viability and ALP activity were observed when cultured SCAP on the surfaces coated with high densities of collagen. TGF-β1 could promote CTGF and α1-integrin gene expression of SCAP in a dose-dependent manner. Furthermore, we cultured SCAP in collagen scaffold with TGF-β1 stimulation and observed the prominent cell proliferation and matrix-like tissue regeneration. When SCAP were cultured in dentin disc scaffold with TGF-β1 stimulation, regenerated dentin-like tissues containing dentinal tubules and calcified nodules were found. Conclusion: SCAP cultured on the collagen matrices with various concentrations showed different biological characteristics like cell attachment, proliferation, and differentiation. Besides, collagen and dentin disc scaffolds with combination of SCAP and TGF-β1 may be practical models in regeneration of dentin-pulp tissues. Further research may be engaged in applying the models in animal studies, and even in clinical practice to repair the teeth with pulp necrosis or immature root formation. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T16:04:46Z (GMT). No. of bitstreams: 1 ntu-102-R99422025-1.pdf: 1505511 bytes, checksum: 0c3855cb70f182f032b1f81e40c8ff05 (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 中文摘要 1
Abstract 3 Chapter I Literature Review 1. Introduction 5 1.1 Stem cells from apical papilla 5 1.2 Transforming growth factor-β superfamily (TGF-βs) and transforming growth factor-β (TGF-β) 8 1.2.1 The Structure and isoforms of TGF-β 8 1.2.2 TGF-β1 in dental-pulp complex 10 1.3 Scaffold 10 1.3.1 Extracellular matrix (ECM) 11 1.3.2 Type I collagen as scaffold 12 Chapter II The Purposes and Hypothesis of the Study 14 Chapter III Materials and Methods 15 3.1 Isolation and culture of SCAP 16 3.2 Surface markers of SCAP 16 3.3 Collagen coating preparation 17 3.4 Cell attachment 17 3.5 Cell viability 18 3.6 Alkaline phosphatase expression in SCAP (ALP stain) 18 3.7 mRNA expression analyzed by RT-PCR 19 3.7.1 Isolation of total RNA 19 3.7.2 RNA quantification 20 3.7.3 Reverse transcription (RT) 21 3.7.4 Polymerase chain reaction (PCR) 21 3.8 Collagen gel incubation 22 3.8.1 Collagen gel preparation 22 3.8.2 Cell seeding 22 3.8.3 Histological analysis 23 3.9 Dentin disc and collagen gel scaffold 23 3.9.1 Dentin disc preparation 23 3.9.2 Cell seeding and histology analysis 24 3.10 Statistical analysis 24 Chapter IV Result 4.1 Morphological observation of SCAP 25 4.2 Surface marker of SCAP 25 4.3 Attachment ability of SCAP on culture wells coated with collagen 25 4.3.1 Morphological changes 25 4.3.2 Determination of attached SCAP on the surfaces coated with collagen 26 4.4 Effects of collagen on cell viability of SCAP - MTT assay 27 4.5 Effect of collagen on ALP activity of SCAP 28 4.6 Effect of TGF-β1 on the CTGF, α1-integrin gene expression of SCAP 28 4.7 The growth characteristics of SCAP in the collagen gels 28 4.8 The growth characteristics of SCAP in the dentin disc supported by collagen 29 Chapter V Discussion Part I: Collagen and SCAP 5.1 Cultured on the collagen may change cell morphology and improve attachment ability of SCAP 30 5.2 The stiffness of collagen matrices may regulate viability of SCAP 32 5.3 The stiffness of collagen matrices may regulate the ALP activity of SCAP 33 Part II: SCAP in scaffolds with TGF-β1 5.4 TGF-β1 up-regulated the gene expression of CTGF and α1-integrin 35 5.5 Tissue regeneration by SCAP cultured in 3D collagen scaffold with TGF-β1 in vitro 37 5.6 Tissue regeneration by SCAP cultured in collagen-supported dentin disc scaffold in combination with TGF-β1 in vitro 39 Chapter IV Conclusion 41 References 43 Figures and tables 50 | |
dc.language.iso | en | |
dc.title | Collagen和TGF-β1對於牙根尖幹細胞行為的影響 | zh_TW |
dc.title | Effect of collagen and TGF-β1 on the behavior of human stem cells from apical papilla | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 張曉華 | |
dc.contributor.oralexamcommittee | 洪善鈴,楊淑芬,張美姬 | |
dc.subject.keyword | 牙根尖細胞,轉型生長因子,膠原蛋白,細胞支架, | zh_TW |
dc.subject.keyword | SCAP,TGF-β1,collagen,scaffold, | en |
dc.relation.page | 71 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2013-06-25 | |
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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