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DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 陳羿貞(Yi-Jane Chen) | |
dc.contributor.author | Lien-Chi Wang | en |
dc.contributor.author | 王簾綺 | zh_TW |
dc.date.accessioned | 2021-06-13T15:47:54Z | - |
dc.date.available | 2013-10-05 | |
dc.date.copyright | 2011-10-05 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-08-10 | |
dc.identifier.citation | Addae-Mensah K. A., Wikswo J. P. (2008) “Measurement techniques for cellular biomechanics in vitro” Exp Biol Med (Maywood). 233(7):792-809.
Basso N., Heersche J. N. M. (2002) “Characteristics of in vitro osteoblastic cell loading models” Bone. 30(2):347-51. Bolcato-Bellemin A. L., et al. (2000) “Expression of mRNAs encoding for α and β integrin subunits, MMPs, and TIMPs in strtched human periodontal ligament and gingival fibroblasts” J Dent Res. 79(9):1712-6. Chen S., et al. (2009) “Runx2, Osx, and DSPP in tooth development” J Dent Res. 88(10):904-9. Ding G., et al. “Effect of cryopreservation on biological and immunological properties of stem cells from apical papilla” J Cell Physiol. 223(2):415-22. D’Souza R.N., et al. (1997) “Gene expression patterns of murine dentin matrix protein 1 (DMP-1) and dentin sialophosphoprotein (DSPP) suggest distinct developmental functions in vivo” J Bone Miner Res. 12(12):2040-9. Han M. J., et al. (2008) “Effect of mechanical tension on the human dental pulp cells” Biotechnol Bioprocess Eng. 13(4):410-417Honda M. J., et al. (2006) “Shear stress facilitateds tissue-engineered odontogenesis” Bone. 39(1):125-33. Huang C. H., et al. (2009) “Interactive effects of mechanical stretching and extracellular matrix proteins on the initiating osteogenic differentiation of human mesenchymal stem cells” J Cell Biochem. 108(6):1263-73. Huang G. T. J., et al. “The hidden treasure in apical papilla: the potential role in pulp/dentin regeneration and bioroot engineering” J Endod. 34(6):645-51. Ikeda E., et al. (2006) “Osteogenic differentiation of human dental papilla mesenchymal cells” Biochem Biophys Res Commun. 342(4):1257-62. Koike M., et al. (2005) “Effects of mechanical strain on proliferation and differentiation of bone marrow stromal cell line ST2” J Bone Miner Metab. 23(3):219-25. Kraft D. C. E., et al. (2010) “Mechnosensitivity of dental pulp stem cells is related to their osteogenic maturity” Eur J Oral Sci. 118(1):29-38. Krishnan V., Davidovitch Z. (2009) “On a path to onfolding the biological mechanisms of orthodontic tooth movement” J Dent Res. 88(7):597-608. Lee S. K., et al. (2010) “Mechanical stress promotes odontoblastic differentiation via the heme oxygenase-1 pathway in human dental pulp cell line” Life Sci. 86(34):107-14. Magne D., et al. (2004) “Development of an odontoblast in vitro model to study dentin mineralization” Connect Tissue Res. 45(2):101-8. Matsuda N., et al. (1998) “Role of epidermal growth factor and its receptor in mechanical stress-induced differentiation of human periodontal ligament cells in vitro” Arch Oral Biol. 43(12):987-97. Miura M., et al. (2003) “SHED: stem cells from human exfoliated deciduous teeth” Proc Natl Acad Sci U S A. 100(10):5807-12. Morsczeck C., et al. (2008) “Somatic stem cells for regenerative dentistry” Clin Oral Investig. 12(2):113-8. Norton L. A., et al. (1995) “A methodical study of shape changes in human oral cells perturbed by a simulated orthodontic strain in vitro” Arch Oral Biol. 40(9):863-72. Park B. W., et al. (2009) “In vitro osteogenic differentiation of cultured human dental papilla-derived cells” J Oral Maxillofac Surg. 67(3):507-14. Qi M. C., et al. (2008) “Mechanical strain induces osteogenic differentiation: Cbfa1 and Ets-1 expression in stretched rat mesenchymal stem cells” Int J Oral Maxillofac Surg. 37(5):453-8. Qin C., et al. (2002) “The expression of dentin sialophosphoprotein gene in bone” J Dent Res. 81(6):392-4. Qin C., O. Baba, W. T. Butler. (2004) “Post-translational modifications of SIBLING proteins and their roles in osteogenesis and dentinogenesis” Crit Rev Oral Biol Med. 15(3):126-36. Rodriguez A. P., et al. (2009) “Influence of the microenvironment on gene and protein expression of odontogenic-like and osteogenic-like cells” Biocell. 33(1):39-47. Saito M., et al. (1991) “Interleukin 1 beta and prostaglandin E are involved in the response of periodontal cells to mechanical stress in vivo and in vitro” Am J Orthod Dentofacial Orthop. 99(3):226-40 Shih I.M., et al. (1999) “The role of CD146 (Mel-CAM) in biology and pathology” J Pathol. 189(1):4-11 Simmons P. J., Torok-Storb B. (1991) “Identification of stromal cell precursors in human bone marrow by a novel monoclonal antibody, STRO-1” Blood. 78(1):55-62. Simon S., et al. (2009) “Molecular characterization of young and mature odontoblasts” Bone. 45(4):693-703. Epub 2009 Jun 23. Sonoyama W., et al. (2006) “Mesenchymal stem cell-mediated functional tooth regeneration in swine” PLoS One. 1:e79. Sonoyama W., 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. Valentin D., et al. (2007) “Mechanical loading down-regumates peroxisome proliferator-activated receptor γ in bone marrow stroma cells and favors osteoblastogenesis at the expense of adipogenesis” Endocrinology. 148(5):2553-62. Visconti L. A., E. H. K. Yen, R. B. Johnson. (2004) “Effect of strain on bone nodule formation by rat osteogenic cells in vitro” Arch Oral Biol. 49(6):485-92. Wei X., et al. (2007) “Expression of mineralization markers in dental pulp cells” J Endod. 33(6):703-8. Yoshino H., et al. (2003) “Mechanical stress induces production of angiogenic regulators in cultured human gingival and periodontal ligament fibroblasts” J Periodontal Res. 38(4):405-10. Yu V., et al. (2009) “Dynamic hydrostatic pressure promotes differentiation of human dental pulp stem cells” Biochem Biophys Res Commun. 386(4):661-5. Zhao Y. H., et al. (2008) “Expression of Osterix in mechanical stress-induced osteogenic differentiation of periodontal ligament cells in vitro” Eur J Oral Sci. 116(3):199-206. Ziros P. G., et al. (2202) “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/123456789/37862 | - |
dc.description.abstract | 隨著生物組織工程研究的發展,研究者已經可以從胚胎或成體組織中分離取得幹細胞;含有幹細胞的牙齒組織包括齒槽骨、牙周韌帶、牙髓、牙濾泡、牙胚等等,由牙齒組織培養幹細胞,相對於胚胎或其它器官而言,則是較不具爭議性、較易取得且傷害較小。體外研究已證實在適當礦化誘導刺激下,齒源幹細胞具有細胞外基質礦化能力,可做為硬組織工程研究發展所需的細胞來源,未來也可能應用在牙齒或齒槽骨缺失的修復。由牙根尖乳突組織分離出來的牙根尖乳突細胞擁有間葉細胞及幹細胞的特性,亦被證實在適當刺激下,具有與骨髓間葉細胞相當的成骨分化能力。回顧文獻中許多研究證實機械力量刺激會促進骨髓間葉幹細胞的成骨分化,但是機械力量刺激對牙根尖乳突幹細胞的生長及成骨/成牙本質分化有何影響,則至今尚未被探討。
牙齒矯正移動時,矯正力對牙周組織造成張力刺激,當牙齒的牙根發育未完全時,則牙根尖乳突組織也須承受張力刺激。本實驗藉由體外實驗模式,以Flexercell system施予週期性張力刺激(3%、0.5 Hz,48小時), 探討張力刺激對牙根尖乳突細胞的形態及排列之影響,以及成骨/成牙本質標記基因或蛋白質表現之變化。 本實驗對於所培養的人類牙根尖乳突細胞進行初步的細胞表面抗原鑑定,結果顯示部份細胞對STRO-1及CD146抗體呈現陽性反應,表示此群細胞內含有間葉細胞及幹細胞的組成。受週期性張力刺激的細胞在形態及細胞骨架排列較具規則性,可知細胞接收外力刺激,將訊息傳入細胞核內進行一連串的基因及蛋白質製造,並表現於形態及排列上,以應對外界環境變化。此外,即時聚合酶鏈鎖反應分析發現受力後6小時及24小時,Cbfa1/Run2(Core-binding factor alpha-1/Runt-related transcription factor-2)、ALP(Alkaline phosphatase)及DSPP(Dentin sialophosphoprotein)基因表現都隨時間增加而上升,但皆未達到統計上的顯著性,週期性張力刺激作用48小時後,Cbfa1、ALP及DSPP基因顯著被向上調控,顯示張力刺激促使牙根尖乳突細胞往成骨/成牙本質分化的方向發展。 本實驗另以西方墨點法觀察Cbfa1/Runx2蛋白質及磷酸化的表現,Cba1/Runx2是成骨分化的重要轉錄因子,也與牙齒硬組織的成熟有關。本實驗結果發現牙根尖乳突細胞受力6小時後,無論是Cbfa1/Runx2總量或其磷酸化之比例,受力組及對照組之間皆無明顯的差異,但在24小時及48小時,受力組的Cbfa1/Runx2總量及磷酸化的程度都較明顯,且在24小時組別達到統計上的顯著性。 本研究首次發現周期性張力刺激可促進人類牙根尖乳突細胞在成骨/成牙本質分化標記基因的向上調控,也說明了受張力的人類牙根尖乳突細胞會往成骨/成牙本質分化的方向發展,但成骨與成牙本質分化的標記往往有所重疊而難以單獨討論,故期許以此研究為基礎,日後能繼續對成骨/成牙本質分化標記及成牙本質分化機制有更多探討。 | zh_TW |
dc.description.abstract | Recent advancements in tissue engineering enable us to discover stem cells from human embryonic and somatic origins. However, because of ethical consideration and risks of operation approaching vital organs, dental tissues such as alveolar bone, periodontal ligaments, dental pulp, dental follicle, and dental papilla, are more easily accessible sources without life-threatened surgical intervention. It was reported that the cells derived from apical papilla (APCs) of developing teeth were capable of osteogenic differentiation as bone marrow stromal cells did while cultured in osteogenic induction. Previous studies showed that mechanical force drive the differentiation of marrow stromal cells into osteogenic lineage. However, there is little understanding of how does the mechanical force affect on osteogenic/dentinogenic differentiation of APCs.
The orthodontic forces applied to an immature tooth might produce the tensional stress to the apical papilla tissue. The aim of this study was to investigate the effects of cyclic mechanical tensional force on the shape, alignment, or the gene or protein expressions of osteogenic/dentinogenic differentiation of the human APCs. By using Flexercell strain unit, we applied the mechanical stimulation with the protocol of 3% elongation at 0.5Hz for 48 hours.. The cultured human APCs were characterized by anti-STRO-1 antibody and anti-CD146 antibody. The stretched APCs were well-oriented in the alignment of cytoskeleton compared to unstretched control. The expressions of Cbfa1/Runx2 (Core-binding factor alpha-1/Runt-related transcription factor-2), ALP (alkaline phosphatase) and DSPP (dentin sialophosphoprotein) genes were slightly, but not significantly elevated in the APCs subjected to cyclic mechanical tensional force for 6-hr and 24-hr. However, the mRNA levels of Cbfa1/Runx2, ALP and DSPP significantly iuncreased in APCs subjected to mechanical stimulation for 48-hr. This results implied that the stretched APCs exhibited the potential to differentiate into osteoblastic/odontoblastic cells. Furthermore, we investigated the protein expression and phosphorylation of Cbfa1/Runx2. Cbfa1/Runx2 is a key transcriptional factor in osteogenic differentiation, and also plays lots role in maturation and formation of dental hard tissue. In the groups with short-term mechanical stretching (6-hr), there was no obvious difference in Cbfa1/Runx2 expression between stretched group and control group. However, the expression of total protein and phosphorylation of Cbfa1/Runx2 significantly increased in the APCs subjected to cyclic mechanical tensional force for 24-hr and 48-hr. Our study was the first to report the upregulation of osteogenic/dentinogenic genes in human APCs by cyclic tensional force. It implied that appropriate mechanical stimulation could promote the osteogenic/dentinogenic differentiation of human APCs. Further studies are needed to discover more the underlying mechanism for controlling the osteogenic/dentinogenic differentiation of human dental apical papilla cells. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T15:47:54Z (GMT). No. of bitstreams: 1 ntu-100-R97422026-1.pdf: 3041985 bytes, checksum: 154e470dcb088b33b15a340c3511f03d (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | 目 錄
口試委員會審定書 ⅰ 誌謝 ⅱ 中文摘要 ⅳ 英文摘要 ⅵ 論文正文 第一章 引言 1 一、人類牙齒的發育 3 二、牙本質生成及成牙本質分化標記 4 三、人類牙根尖乳突細胞 5 四、機械性張力刺激 7 五、細胞受力文獻回顧 8 第二章 實驗目的 11 第三章 材料與方法 一、細胞培養 12 二、流式細胞儀分析 12 三、張力系統及週期性張力刺激 13 四、細胞形態及排列之觀察 14 五、肌動蛋白細胞骨架及細胞焦點附著染色 14 六、基因表現之觀察 15 七、Cbfa1/Runx2蛋白質表現之觀察 17 八、鹼性磷酸酶染色 19 九、統計分析 19 第四章 結果 一、人類牙根尖乳突細胞之形態觀察 20 二、流式細胞儀分析人類牙根尖乳突細胞 20 三、細胞受週期性張力刺激後形態、排列及數量之變化 20 四、細胞受週期性張力刺激後細胞骨架之變化 21 五、Real-time PCR分析人類牙根尖乳突細胞受週期性張力刺激後成骨/成牙本質基因之表現 22 六、西方墨點法分析人類牙根尖乳突細胞受週期性張力刺激後而Cbfa1/Runx2蛋白質表現 23 七、人類牙根尖乳突細胞受週期性張力刺激後鹼性磷酸酶的表現 24 第五章 討論 一、人類牙根尖乳突細胞之形態觀察 25 二、牙根尖乳突細胞之鑑定 26 三、週期性張力刺激 27 四、細胞受週期性張力刺激後形態、排列及數量之變化 28 五、促進人類牙根尖乳突細胞分化為成骨母細胞/成牙本質母細胞之討論 30 六、週期性張力刺激對人類牙根尖乳突細胞分化的機制 35 第六章 結論 37 第七章 未來研究方向 38 參考文獻 40 圖 目 錄 圖1 牙根尖乳突組織之組織結構 45 圖2 早期張伸設備 46 圖3 特製張伸設備 47 圖4 本實驗使用之張伸設備Flexercell system 48 圖5 流式細胞儀分析人類牙根尖乳突細胞。 49 圖6 細胞受週期性張力刺激之數量變化 51 圖7 由即時聚合酶鏈鎖反應的分析結果,探討細胞受週期性張力刺激後成骨/成牙本質基因表現 - Cbfa1/Runx2。 53 圖8 由即時聚合酶鏈鎖反應的分析結果,探討細胞受週期性張力刺激後成骨/成牙本質基因表現 - ALP 55 圖9 由即時聚合酶鏈鎖反應的分析結果,探討細胞受週期性張力刺激後成骨/成牙本質基因表現 -OCN。 57 圖10 由即時聚合酶鏈鎖反應的分析結果,探討細胞受週期性張力刺激後成骨/成牙本質基因表現 -DSPP 59 圖11 西方墨點法分析Cbfa1/Runx2蛋白質、磷酸化蛋白質含量及磷酸化蛋白比例 61 圖12 實驗總結 63 表 目 錄 表1 本實驗於即時聚合酶鏈鎖反應所使用引子 64 表2 人類牙根尖乳突細胞之形態觀察 65 表3 細胞受週期性張力刺激之形態觀察 67 表4 細胞受週期性張力刺激後細胞骨架之變化 70 表5 細胞受週期性張力刺激後細胞骨架之變化 72 表6 鹼性磷酸酶染色比較張力刺激後ALP的變化 73 | |
dc.language.iso | zh-TW | |
dc.title | 周期性張力刺激對人類牙根尖乳突細胞生長及分化之影響 | zh_TW |
dc.title | Effects of cyclic tensional force on growth and differentiation of human dental apical papilla cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 鄭景暉(Jiiang-Huei Jeng) | |
dc.contributor.oralexamcommittee | 張宏博(Hong-Po Chang),李勝揚(Sheng-Yang Lee) | |
dc.subject.keyword | 人類牙根尖乳突細胞,週期性張力刺激,成骨/成牙本質分化, | zh_TW |
dc.subject.keyword | Human apical papilla cell,Cyclic mechanical tensional force,Osteogenic/dentinogenic differetiation, | en |
dc.relation.page | 73 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-08-10 | |
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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