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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 楊台鴻 | |
dc.contributor.author | Min-Chun Chung | en |
dc.contributor.author | 鍾明均 | zh_TW |
dc.date.accessioned | 2021-06-08T06:56:54Z | - |
dc.date.copyright | 2009-07-28 | |
dc.date.issued | 2009 | |
dc.date.submitted | 2009-07-21 | |
dc.identifier.citation | Ahn, K. M., J. H. Lee, et al. (2006). 'Fabrication of myomucosal flap using tissue-engineered bioartificial mucosa constructed with oral keratinocytes cultured on amniotic membrane.' Artif Organs 30(6): 411-23.
Alpar, B., G. Leyhausen, et al. (2000). 'Compatibility of resorbable and nonresorbable guided tissue regeneration membranes in cultures of primary human periodontal ligament fibroblasts and human osteoblast-like cells.' Clin Oral Investig 4(4): 219-25. Avery, J. K., Steele P.F., Avery NBFA., editors. (2002). 'Oral Development and Histology.' 3rd. New Youk, USA: Thieme Medical Publishers. pp. 72-106 Bartold, P. M., B. Reinboth, et al. (1990). 'Proteoglycans of bovine cementum: isolation and characterization.' Matrix 10(1): 10-9. Beck, G. R., Jr., E. C. Sullivan, et al. (1998). 'Relationship between alkaline phosphatase levels, osteopontin expression, and mineralization in differentiating MC3T3-E1 osteoblasts.' J Cell Biochem 68(2): 269-80. Benson-Martin, J., P. Zammaretti, et al. (2006). 'The Young's modulus of fetal preterm and term amniotic membranes.' Eur J Obstet Gynecol Reprod Biol 128(1-2): 103-7. Bouvier, M., A. Joffre, et al. (1990). 'In vitro mineralization of a three-dimensional collagen matrix by human dental pulp cells in the presence of chondroitin sulphate.' Arch Oral Biol 35(4): 301-9. Capeans, C., A. Pineiro, et al. (2003). 'Amniotic membrane as support for human retinal pigment epithelium (RPE) cell growth.' Acta Ophthalmol Scand 81(3): 271-7. Carrano, A. V., J. W. Gray, et al. (1979). 'Measurement and purification of human chromosomes by flow cytometry and sorting.' Proc Natl Acad Sci U S A 76(3): 1382-4. Casey, M. L. and P. C. MacDonald (1996). 'Interstitial collagen synthesis and processing in human amnion: a property of the mesenchymal cells.' Biol Reprod 55(6): 1253-60. Chen, R. S., Y. J. Chen, et al. (2007). 'Cell-surface interactions of rat tooth germ cells on various biomaterials.' J Biomed Mater Res A 83(1): 241-8. Chuck, R. S., J. M. Graff, et al. (2004). 'Biomechanical characterization of human amniotic membrane preparations for ocular surface reconstruction.' Ophthalmic Res 36(6): 341-8. Cooper, L. J., S. Kinoshita, et al. (2005). 'An investigation into the composition of amniotic membrane used for ocular surface reconstruction.' Cornea 24(6): 722-729. Covas, D. T., R. A. Panepucci, et al. (2008). 'Multipotent mesenchymal stromal cells obtained from diverse human tissues share functional properties and gene-expression profile with CD146+ perivascular cells and fibroblasts.' Exp Hematol 36(5): 642-54. de Bernard, B. (1982). 'Glycoproteins in the local mechanism of calcification.' Clin Orthop Relat Res(162): 233-44. Dua, H. S., J. A. Gomes, et al. (2004). 'The amniotic membrane in ophthalmology.' Surv Ophthalmol 49(1): 51-77. Duailibi, M. T., S. E. Duailibi, et al. (2004). 'Bioengineered teeth from cultured rat tooth bud cells.' J Dent Res 83(7): 523-8. Ducy, P., R. Zhang, et al. (1997). 'Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation.' Cell 89(5): 747-54. Espana, E. M., H. He, et al. (2003). 'Human keratocytes cultured on amniotic membrane stroma preserve morphology and express keratocan.' Invest Ophthalmol Vis Sci 44(12): 5136-41. Favata, M. F., K. Y. Horiuchi, et al. (1998). 'Identification of a novel inhibitor of mitogen-activated protein kinase kinase.' J Biol Chem 273(29): 18623-32. Franceschi, R. T. (1999). 'The developmental control of osteoblast-specific gene expression: role of specific transcription factors and the extracellular matrix environment.' Crit Rev Oral Biol Med 10(1): 40-57. Franceschi, R. T., G. Xiao, et al. (2003). 'Multiple signaling pathways converge on the Cbfa1/Runx2 transcription factor to regulate osteoblast differentiation.' Connect Tissue Res 44 Suppl 1: 109-16. Frau, S., J. Bernadou, et al. (1997). 'Nuclease activity and binding characteristics of a cationic 'manganese porphyrin-bis(benzimidazole) dye (Hoechst 33258)' conjugate.' Bioconjug Chem 8(2): 222-31. Ge, C., G. Xiao, et al. (2007). 'Critical role of the extracellular signal-regulated kinase-MAPK pathway in osteoblast differentiation and skeletal development.' J Cell Biol 176(5): 709-18. Goldberg, M. and J. J. Lasfargues (1995). 'Pulpo-dentinal complex revisited.' J Dent 23(1): 15-20. Griffiths, G. S., A. M. Moulson, et al. (1998). 'Evaluation of osteocalcin and pyridinium crosslinks of bone collagen as markers of bone turnover in gingival crevicular fluid during different stages of orthodontic treatment.' J Clin Periodontol 25(6): 492-8. Gronthos, S., S. E. Graves, et al. (1994). 'The STRO-1+ fraction of adult human bone marrow contains the osteogenic precursors.' Blood 84(12): 4164-73. Gronthos, S., M. Mankani, et al. (2000). 'Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo.' Proc Natl Acad Sci U S A 97(25): 13625-30. Grueterich, M. and S. C. Tseng (2002). 'Human limbal progenitor cells expanded on intact amniotic membrane ex vivo.' Arch Ophthalmol 120(6): 783-90. Hao, J., S. Shi, et al. (1997). 'Mineralized nodule formation by human dental papilla cells in culture.' Eur J Oral Sci 105(4): 318-24. Harada, M., N. Udagawa, et al. (1986). 'Inorganic pyrophosphatase activity of purified bovine pulp alkaline phosphatase at physiological pH.' J Dent Res 65(2): 125-7. Harris, R. J. (1998). 'Root coverage with a connective tissue with partial thickness double pedicle graft and an acellular dermal matrix graft: a clinical and histological evaluation of a case report.' J Periodontol 69(11): 1305-11. Harris, S. E., L. F. Bonewald, et al. (1994). 'Effects of transforming growth factor beta on bone nodule formation and expression of bone morphogenetic protein 2, osteocalcin, osteopontin, alkaline phosphatase, and type I collagen mRNA in long-term cultures of fetal rat calvarial osteoblasts.' J Bone Miner Res 9(6): 855-63. Higuchi, C., A. Myoui, et al. (2002). 'Continuous inhibition of MAPK signaling promotes the early osteoblastic differentiation and mineralization of the extracellular matrix.' J Bone Miner Res 17(10): 1785-94. Hitomi, K., Y. Torii, et al. (1992). 'Increase in the activity of alkaline phosphatase by L-ascorbic acid 2-phosphate in a human osteoblast cell line, HuO-3N1.' J Nutr Sci Vitaminol (Tokyo) 38(6): 535-44. Ikeda, E., M. Hirose, et al. (2006). 'Osteogenic differentiation of human dental papilla mesenchymal cells.' Biochem Biophys Res Commun 342(4): 1257-62. Inanc, B., A. E. Elcin, et al. (2006). 'Osteogenic induction of human periodontal ligament fibroblasts under two- and three-dimensional culture conditions.' Tissue Eng 12(2): 257-66. Ivanovski, S., H. Li, et al. (2001). 'Expression of bone associated macromolecules by gingival and periodontal ligament fibroblasts.' J Periodontal Res 36(3): 131-41. Julien, M., D. Magne, et al. (2007). 'Phosphate stimulates matrix Gla protein expression in chondrocytes through the extracellular signal regulated kinase signaling pathway.' Endocrinology 148(2): 530-7. Kamalia, N., C. A. McCulloch, et al. (1992). 'Dexamethasone recruitment of self-renewing osteoprogenitor cells in chick bone marrow stromal cell cultures.' Blood 79(2): 320-6. Kamata, N., R. Fujimoto, et al. (2004). 'Immortalization of human dental papilla, dental pulp, periodontal ligament cells and gingival fibroblasts by telomerase reverse transcriptase.' J Oral Pathol Med 33(7): 417-23. Kandavel, G. R. and R. S. Chuck (2005). 'Staining properties of deepithelialized human amniotic membrane.' Cornea 24(7): 853-6. Kasperk, C., U. Schneider, et al. (1995). 'Differential effects of glucocorticoids on human osteoblastic cell metabolism in vitro.' Calcif Tissue Int 57(2): 120-6. Kasugai, S., S. Shibata, et al. (1993). 'Characterization of a system of mineralized-tissue formation by rat dental pulp cells in culture.' Arch Oral Biol 38(9): 769-77. Kemoun, P., S. Laurencin-Dalicieux, et al. (2007). 'Localization of STRO-1, BMP-2/-3/-7, BMP receptors and phosphorylated Smad-1 during the formation of mouse periodontium.' Tissue Cell 39(4): 257-66. Kim, Y. J., R. L. Sah, et al. (1988). 'Fluorometric assay of DNA in cartilage explants using Hoechst 33258.' Anal Biochem 174(1): 168-76. Koide, N., K. Sakaguchi, et al. (1990). 'Formation of multicellular spheroids composed of adult rat hepatocytes in dishes with positively charged surfaces and under other nonadherent environments.' Exp Cell Res 186(2): 227-35. Komori, T. (2002). '[Cbfa1/Runx2, an essential transcription factor for the regulation of osteoblast differentiation].' Nippon Rinsho 60 Suppl 3: 91-7. Kono, S. J., Y. Oshima, et al. (2007). 'Erk pathways negatively regulate matrix mineralization.' Bone 40(1): 68-74. Lai, C. F., L. Chaudhary, et al. (2001). 'Erk is essential for growth, differentiation, integrin expression, and cell function in human osteoblastic cells.' J Biol Chem 276(17): 14443-50. Lallier, T. E., A. Spencer, et al. (2005). 'Transcript profiling of periodontal fibroblasts and osteoblasts.' J Periodontol 76(7): 1044-55. Langer, R. and J. P. Vacanti (1993). 'Tissue engineering.' Science 260(5110): 920-6. Langer, R. and J. P. Vacanti (1995). 'Artificial organs.' Sci Am 273(3): 130-3. Lekic, P., J. Sodek, et al. (1996). 'Osteopontin and bone sialoprotein expression in regenerating rat periodontal ligament and alveolar bone.' Anat Rec 244(1): 50-8. Lian, J. B. and G. S. Stein (1995). 'Development of the osteoblast phenotype: molecular mechanisms mediating osteoblast growth and differentiation.' Iowa Orthop J 15: 118-40. Liu, Y., D. A. Peterson, et al. (1997). 'Mechanism of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction.' J Neurochem 69(2): 581-93. Longui, C. A., M. C. Santos, et al. (2005). 'Antiproliferative and apoptotic potencies of glucocorticoids: nonconcordance with their antiinflammatory and immunosuppressive properties.' Arq Bras Endocrinol Metabol 49(3): 378-83. Luan, X., Y. Ito, et al. (2006). 'Dental follicle progenitor cell heterogeneity in the developing mouse periodontium.' Stem Cells Dev 15(4): 595-608. Lynch, M. P., J. L. Stein, et al. (1995). 'The influence of type I collagen on the development and maintenance of the osteoblast phenotype in primary and passaged rat calvarial osteoblasts: modification of expression of genes supporting cell growth, adhesion, and extracellular matrix mineralization.' Exp Cell Res 216(1): 35-45. Malak, T. M., C. D. Ockleford, et al. (1993). 'Confocal immunofluorescence localization of collagen types I, III, IV, V and VI and their ultrastructural organization in term human fetal membranes.' Placenta 14(4): 385-406. Mariotti, A. (1993). 'The extracellular matrix of the periodontium: dynamic and interactive tissues.' Periodontol 2000 3: 39-63. Morsczeck, C., W. Gotz, et al. (2005). 'Isolation of precursor cells (PCs) from human dental follicle of wisdom teeth.' Matrix Biol 24(2): 155-65. Nakashima, M. and A. H. Reddi (2003). 'The application of bone morphogenetic proteins to dental tissue engineering.' Nat Biotechnol 21(9): 1025-32. Nohutcu, R. M., L. K. McCauley, et al. (1997). 'Expression of extracellular matrix proteins in human periodontal ligament cells during mineralization in vitro.' J Periodontol 68(4): 320-7. Otto, F., A. P. Thornell, et al. (1997). 'Cbfa1, a candidate gene for cleidocranial dysplasia syndrome, is essential for osteoblast differentiation and bone development.' Cell 89(5): 765-71. Owen, T. A., M. Aronow, et al. (1990). 'Progressive development of the rat osteoblast phenotype in vitro: reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix.' J Cell Physiol 143(3): 420-30. Park, B. W., Y. S. Hah, et al. (2009). 'In vitro osteogenic differentiation of cultured human dental papilla-derived cells.' J Oral Maxillofac Surg 67(3): 507-14. Parry, S. and J. F. Strauss, 3rd (1998). 'Premature rupture of the fetal membranes.' N Engl J Med 338(10): 663-70. Peter, S. J., C. R. Liang, et al. (1998). 'Osteoblastic phenotype of rat marrow stromal cells cultured in the presence of dexamethasone, beta-glycerolphosphate, and L-ascorbic acid.' J Cell Biochem 71(1): 55-62. Pruyne, D. and A. Bretscher (2000). 'Polarization of cell growth in yeast.' J Cell Sci 113 ( Pt 4): 571-85. Quarles, L. D., D. A. Yohay, et al. (1992). 'Distinct proliferative and differentiated stages of murine MC3T3-E1 cells in culture: an in vitro model of osteoblast development.' J Bone Miner Res 7(6): 683-92. Rinastiti, M., Harijadi, et al. (2006). 'Histological evaluation of rabbit gingival wound healing transplanted with human amniotic membrane.' Int J Oral Maxillofac Surg 35(3): 247-51. Rodriguez-Ares, M. T., R. Tourino, et al. (2004). 'Multilayer amniotic membrane transplantation in the treatment of corneal perforations.' Cornea 23(6): 577-83. Sawa, Y., Y. Yamaoka, et al. (2004). 'Reduction of alkaline phosphatase activity in aged human osteogenic periodontal ligament fibroblasts exhibiting short telomeres.' Cell Tissue Res 315(3): 331-7. Seibel, M. J., Robins, S. P., and Bilezikian, J. P. (1999). 'Acid and Alkaline phosphatases. Dynamics of Bone and Cartilage Metacollism.' New York: Academic Press. pp. 127-135 Seo, B. M., M. Miura, et al. (2004). 'Investigation of multipotent postnatal stem cells from human periodontal ligament.' Lancet 364(9429): 149-55. Shiga, M., Y. L. Kapila, et al. (2003). 'Ascorbic acid induces collagenase-1 in human periodontal ligament cells but not in MC3T3-E1 osteoblast-like cells: potential association between collagenase expression and changes in alkaline phosphatase phenotype.' J Bone Miner Res 18(1): 67-77. Shih, I. M. (1999). 'The role of CD146 (Mel-CAM) in biology and pathology.' J Pathol 189(1): 4-11. Simmons, P. J. and B. Torok-Storb (1991). 'Identification of stromal cell precursors in human bone marrow by a novel monoclonal antibody, STRO-1.' Blood 78(1): 55-62. Smith, P. K., R. I. Krohn, et al. (1985). 'Measurement of protein using bicinchoninic acid.' Anal Biochem 150(1): 76-85. Sonoyama, W., Y. Liu, et al. (2008). 'Characterization of the apical papilla and its residing stem cells from human immature permanent teeth: a pilot study.' J Endod 34(2): 166-71. Springer, T. A. (1996). 'Immunopreceipitation.' In Current Proposols in Immunology (J.E. Coligan, A.M. Kruisbeck, D.H. Margulies, E.M. Shevach, and W. Strober, eds.) pp.8.3.1-8.3.11 John Wiley & Sons, New York. Stock, M. and F. Otto (2005). 'Control of RUNX2 isoform expression: the role of promoters and enhancers.' J Cell Biochem 95(3): 506-17. Subrahmanyam, M. (1995). 'Amniotic membrane as a cover for microskin grafts.' Br J Plast Surg 48(7): 477-8. Sudha, B., S. Jasty, et al. (2009). 'Signal transduction pathway involved in the ex vivo expansion of limbal epithelial cells cultured on various substrates.' Indian J Med Res 129(4): 382-9. Tabata, Y. (2009). 'Biomaterial technology for tissue engineering applications.' J R Soc Interface 6 Suppl 3: S311-24. Taira, M., H. Nakao, et al. (2003). 'Effects of two vitamins, two growth factors and dexamethasone on the proliferation of rat bone marrow stromal cells and osteoblastic MC3T3-E1 cells.' J Oral Rehabil 30(7): 697-701. Takeuchi, Y., M. Suzawa, et al. (1997). 'Differentiation and transforming growth factor-beta receptor down-regulation by collagen-alpha2beta1 integrin interaction is mediated by focal adhesion kinase and its downstream signals in murine osteoblastic cells.' J Biol Chem 272(46): 29309-16. Ten Cate (1998). 'Oral Histology: Development, structure, and Function.' Mosby. Thesleff, I. (1986). 'Dental papilla cells in culture. Comparison of morphology, growth and collagen synthesis with two other dental-related embryonic mesenchymal cell populations.' Cell Differ 18(3): 189-98. Thesleff, I. (2003). 'Epithelial-mesenchymal signalling regulating tooth morphogenesis.' J Cell Sci 116(Pt 9): 1647-8. Thesleff, I. and P. Sharpe (1997). 'Signalling networks regulating dental development.' Mechanisms of Development 67(2): 111-123. Tseng, S. C., D. Q. Li, et al. (1999). 'Suppression of transforming growth factor-beta isoforms, TGF-beta receptor type II, and myofibroblast differentiation in cultured human corneal and limbal fibroblasts by amniotic membrane matrix.' J Cell Physiol 179(3): 325-35. Tsuchiya, S., M. J. Honda, et al. (2008). 'Collagen type I matrix affects molecular and cellular behavior of purified porcine dental follicle cells.' Cell Tissue Res 331(2): 447-59. Wagshall, E., Z. Lewis, et al. (2002). 'Acellular dermal matrix allograft in the treatment of mucogingival defects in children: illustrative case report.' ASDC J Dent Child 69(1): 39-43, 11. Xie, J. M., W. D. Tian, et al. (2005). '[Culture and characteristics of human dental papilla cells in vitro].' Hua Xi Kou Qiang Yi Xue Za Zhi 23(3): 187-90. Young, C. S., S. Terada, et al. (2002). 'Tissue engineering of complex tooth structures on biodegradable polymer scaffolds.' J Dent Res 81(10): 695-700. Zhang, Y., M. Ni, et al. (2003). 'Calcium phosphate-chitosan composite scaffolds for bone tissue engineering.' Tissue Eng 9(2): 337-45. Zhao, M., G. Xiao, et al. (2002). 'Bone morphogenetic protein 2 induces dental follicle cells to differentiate toward a cementoblast/osteoblast phenotype.' J Bone Miner Res 17(8): 1441-51. Ziros, P. G., A. P. Gil, et al. (2002). 'The bone-specific transcriptional regulator Cbfa1 is a target of mechanical signals in osteoblastic cells.' J Biol Chem 277(26): 23934-41. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/25907 | - |
dc.description.abstract | 人類羊膜是一天然性生物材料,目前已有臨床報告指出,羊膜可被成功應用為貼附材料以促進傷口癒合、眼角膜重建等;隨著幹細胞研究的發展,利用組織工程修復口腔組織缺損已不是遙不可及的夢想,近年來已有研究證實牙根尖乳突細胞群在適當的誘導下能表現多重分化的潛能,因此,本研究的目的在探討羊膜作為口腔細胞生長基材的情形,以及羊膜對於牙根尖乳突細胞生長及功能的影響,以評估將羊膜應用於口腔內組織修復的可能性。
為探討羊膜是否適合口腔細胞之附著及生長,我們分別以牙根尖乳突細胞、牙齦纖維母細胞、牙周韌帶纖維母細胞、牙髓細胞與骨髓間葉幹細胞等五種細胞進行基本測試,測試各種細胞在鋪植後不同時間點(1、3、5、7天)的MTT活性,比較羊膜基材用於口腔細胞培養的適合性。將各種細胞培養在經去細胞處理的羊膜基底層與實質層上,對其貼附、活性及形態做探討,從結果中發現,牙根尖乳突細胞在羊膜基材上的貼附與活性並不如tissue culture polystrene(TCPS),牙髓細胞與骨髓間葉幹細胞在生長活性上也有類似的結果,但對於牙齦纖維母細胞及牙周韌帶纖維母細胞來說,羊膜是有利於細胞生長的基材。 本研究對於培養自發育中第三大臼齒根尖牙乳突部位之細胞進行初步鑑定,利用STRO-1與CD146抗體做免疫螢光染色及流式細胞儀分析,探討牙根尖乳突細胞是否具有間葉幹細胞的特性,結果顯示部分牙根尖乳突細胞群對此兩種抗體的表現為陽性,可說明本研究所培養測試之人類牙根尖乳突細胞為具有間葉細胞特性的前驅細胞。在促進細胞基質礦化之培養液中,人類牙根尖乳突細胞的鹼性磷酸酶活性表現隨培養時間增長而上升,顯示其具有類似成骨細胞分化的基本特性;而相對於TCPS,生長在羊膜基材上的牙根尖乳突細胞都具有較高的ALP活性,細胞外基質礦化程度也較強,且有提早促進COL I(collagen type I)、Cbfa1(Core-binding factor-1)、ALP(alkaline phospatase)及OC(osteocalcin)基因表現的效應。 本研究更進一步利用ERK1/2抑制劑(U0126)探討牙根尖乳突細胞在羊膜上之分化促進是否與ERK1/2訊息傳導路徑有關,結果証明牙根尖乳突細胞的分化確實與ERK1/2有關,但關於羊膜基質對於細胞的成骨分化能力之促進作用,ERK或許並不是最主要的角色。 | zh_TW |
dc.description.abstract | Human amniotic membrane (AM) is a nature biomaterial which has been successfully used in cornea reconstruction and as a biological dressing for wounds. Recently, it was reported that apical papilla cells were multipotent and their ability of differentiating into specific lineage could be induced in an appropriate environment. Therefore, the purpose of this study was to investigate the possibility of applying human amnions for oral tissue repair by evaluating the growth and function of human apical papilla cells cultured in the de-cellular human amniotic matrix.
In addition to apical papilla cells, several kinds of dental mesenchymal cells were used to check the feasibility of de-cellular AM matrix for cellular adhesion and growth, including dental pulp cells, gingival fibroblasts, periodontal ligament fibroblasts, and bone marrow mesenchymal stem cells. The apical papilla cells were cultured from human apical papilla of developing third molar extracted due to orthodontic reason. The dental progenitor cells in the mixed population cultured from apical papilla was characterized by using anti- STRO-1 and anti- CD146 antibody for immunofluorescence stain and flow cytometry. The osteogenic differentiation of apical papilla cells grown on different substrates (tissue culture plate (TCPS), basement membrane of AM, and collagenous matrix of AM) was investigated by measuring the alkaline phosphotase activity and extra-cellular matrix mineralization revealed by von Kossa stain. The mRNA expressions of osteoblast-related genes (COL I, Cbfa1, ALP, and OC) were checked by using RT-PCR technique. Protein expression and phosphorylation of RUNX-2 were detected via the procedures of imunoprecipitation and western blot. Moreover, we used U0126, an inhibitor of ERK1/2 pathway, to check if the ERK 1/2 pathway is involved in the osteogenic differentiation of apical papilla cells grown on AM. The result showed that the adhesion and proliferation of apical papilla cells cultured on amniotic membrane were not as good as those on TCPS. The similar difference between amniotic membrane and TCPS for cellular growth was noted for dental pulp cells and bone marrow mesenchymal stem cells. However, amniotic membrane was a good matrix to support the growth of gingival fibroblasts and periodontal ligament fibroblasts. The osteoblastic phenotype of human apical papilla cells was demonstrated by their progressive increase of ALP activity and extracellular matrix mineralization in long-term culture. Compared to TCPS, AM could promote osteogenic differentiation of apical papilla cells. The mRNA expression of COL I, Cbfa1, ALP, and OC was also up-regulated in apical papilla cells grown on AM. Moreover, the osteogenic differentiation of apical papilla cells, either on TCPS or on AM, was significantly down-regulated in the presence of U0126. It implied that ERK1/2 signaling pathway was activated in the osteogenic differentiation of apical papilla cells, followed by up-regulating the expression and phosphorylation of RUNX-2, subsequently increasing ALP activity and mineralized matrix deposition. However, the promoting effect of AM on the osteogenic differentiation of apical papilla cells may not directly involve the ERK1/2 signaling pathway. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T06:56:54Z (GMT). No. of bitstreams: 1 ntu-98-R96548016-1.pdf: 4605839 bytes, checksum: 2e286794efe30e674c55f1fa449fb7b4 (MD5) Previous issue date: 2009 | en |
dc.description.tableofcontents | 摘要 i
Abstract iii 目錄 v 表目錄 ix 圖目錄 xi 第一章 緒論 1 1-1 研究背景 1 1-1-1 牙周組織與缺損 1 1-1-2 牙周修復與牙齒再生 2 1-1-3 組織工程(tissue engineering)原理 3 1-2 研究目的 4 第二章 文獻回顧 5 2-1 牙齒發育及牙囊組織的特徵 5 2-2 牙根尖乳突細胞的特性 6 2-3 細胞分化與成骨指標蛋白的基因表現 8 2-3-1 第一型膠原蛋白(Collagen type I,COL I) 9 2-3-2 Cbfa1/Runx2(Core-binding factor-1/Runt-related transcription factor-2) 9 2-3-3 鹼性磷酸酶(Alkaline phosphatase,ALP) 10 2-3-4 骨鈣蛋白(Osteocalcin,OC) 10 2-4 人類羊膜的組織形態 10 2-5 人類羊膜於體外培養實驗的模式 11 2-6 應用羊膜於促進傷口癒合的模式 12 第三章 實驗材料與方法 15 3-1 細胞培養(cell culture) 15 3-1-1 人類牙根尖乳突細胞(Human Apical papilla cells,DF) 15 3-1-2 人類牙周韌帶纖維細胞(Human Periodontal Ligament Fibroblasts,PF) 16 3-1-3 人類牙齦纖維細胞(Human Gingival Fibroblasts,GF) 16 3-1-4 人類牙髓細胞(Human Dental Pulp cells,DP) 16 3-1-5 人類骨髓幹細胞(Human Bone Marrow Stem cell,BMSC) 17 3-2 去細胞羊膜之配製 17 3-3 礦化培養液(Mineralizing Medium)之配製 18 3-4 細胞生長測試(MTT assay) 18 3-5 免疫螢光染色法(immunofluorescence stain) 19 3-5-1 STRO-1與CD146染色 19 3-5-2 肌動蛋白細胞骨架染色(Actin Cytoskeleton Staining) 20 3-5-3 Hoechst 33258染色 20 3-6 流式細胞儀分析(Flow cytometry) 21 3-7 促進基質礦化實驗之成骨基因表現 21 3-7-1 RNA萃取 21 3-7-2 反轉錄聚合酶連鎖反應(Reverse Transcription-Polymerase Chain Reaction,RT-PCR) 22 3-7-3 瓊脂凝膠電泳分析(Agarose gel electrophoresis) 23 3-8 鹼性磷酸酶活性(Alkaline Phosphatase Activity)測定 23 3-9 鈣化節點染色法(von Kossa stain) 25 3-10 西方墨點法(Western Blot) 25 3-10-1 細胞內蛋白質分離 26 3-10-2 免疫沉澱(immunoprecipitation,IP) 26 3-10-3 西方墨點法(Western Blot) 27 3-11 統計分析 28 第四章 結果 29 4-1 MTT活性測試 29 4-1-1 人類牙根尖乳突細胞培養於TCPS及羊膜基材之生長情形 29 4-1-2 人類牙周韌帶纖維母細胞培養於TCPS及羊膜基材之生長情形 29 4-1-3 人類牙齦纖維母細胞培養於TCPS及羊膜基材之生長情形 30 4-1-4 人類牙髓細胞培養於TCPS及羊膜基材之生長情形 30 4-1-5 人類骨髓間葉幹細胞培養於TCPS及羊膜基材之生長情形 30 4-2 牙根尖乳突細胞形態與鑑定 30 4-2-1 牙根尖乳突細胞形態觀察 30 4-2-2 牙根尖乳突細胞鑑定 31 4-3 牙根尖乳突細胞培養於TCPS與羊膜的情形 31 4-3-1 細胞貼附的形態 31 4-3-2 Hoechst33258染色結果 32 4-4 成骨基因表現 33 4-4-1 COL I 基因表現 33 4-4-2 Cbfa1 基因表現 33 4-4-3 ALP基因表現 34 4-4-4 OC基因表現 34 4-5 鹼性磷酸酶活性測定 34 4-6 鈣化節點染色法觀察 35 4-7 Cbfa1/Runx2蛋白質表現 36 4-8 U0126作用的影響 36 4-8-1 不同濃度之U0126對於牙根尖乳突細胞生長活性之測試 37 4-8-2 不同濃度之U0126對於牙根尖乳突細胞之鹼性磷酸酶活性測試 37 4-8-3 U0126在不同材料上對ERK1/2磷酸化的影響 37 4-8-4 U0126在不同材料上對成骨化基因的影響 38 4-8-5 U0126在不同材料上之鹼性磷酸酶活性差異 38 4-8-6 U0126在不同材料上之鈣化節點染色法 39 4-8-7 U0126在不同材料上之Cbfa1/Runx2蛋白磷酸化的影響 39 第五章 討論 40 5-1 牙根尖乳突細胞形態 40 5-2 牙根尖乳突細胞鑑定 40 5-3 羊膜對牙根尖乳突細胞生長的影響 41 5-4 促進牙根尖乳突細胞基質礦化的研究 42 5-5 人類羊膜對牙根尖乳突細胞成骨分化的影響 45 5-5-1 COL I(collagen type I) 46 5-5-2 Cbfa1/Runx2(Core-binding factor-1/Runt-related transcription factor-2) 46 5-5-3 ALP(alkaline phosphase) 47 5-5-4 OC(osteocalcin) 48 5-6 牙根尖乳突細胞鈣化沉積物的產生 49 5-7 羊膜影響牙根尖乳突細胞分化能力的機制 50 第六章 結論 53 第七章 未來研究方向 55 參考文獻 56 附錄(表與圖) 66 | |
dc.language.iso | zh-TW | |
dc.title | 人類牙根尖乳突細胞培養於羊膜基材之研究 | zh_TW |
dc.title | Study of human apical papilla cells cultured on amniotic membrane matrix | en |
dc.type | Thesis | |
dc.date.schoolyear | 97-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 陳羿貞 | |
dc.contributor.oralexamcommittee | 林俊彬,鄭景暉 | |
dc.subject.keyword | 人類羊膜,牙根尖乳突細胞,成骨分化, | zh_TW |
dc.subject.keyword | human amniotic membrane,apical papilla cells,osteogenic differentiation, | en |
dc.relation.page | 94 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2009-07-21 | |
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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