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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 | Bo-Hao Zhong | en |
dc.contributor.author | 仲柏澔 | zh_TW |
dc.date.accessioned | 2021-07-11T15:28:22Z | - |
dc.date.available | 2021-10-09 | |
dc.date.copyright | 2018-10-09 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-08-18 | |
dc.identifier.citation | AAE. (2016). Glossary of endodontic terms. Chicago: American Association of Endodontists.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78908 | - |
dc.description.abstract | 實驗目的:褪黑素因具有多種不同的生物學功能,已經被廣泛應用於幹細胞和組織工程等研究領域。而根據先前的研究,人類牙根尖細胞 (apical papilla cells) 顯示出具有幹細胞的特徵跟能力,並被稱為根尖乳突幹細胞 (Stem cells from the apical papilla, SCAP)。本研究的目的是想探討褪黑素對牙根尖細胞的影響以及PKA和PKC信號通路的作用。實驗方法:本實驗用不同濃度的褪黑素來刺激與培養牙根尖細胞,部分組別加入H89 (PKA抑制劑) 或H7 (PKC抑制劑) 做預處理。再以顯微鏡觀察細胞形態,並用MTT 做細胞存活率分析。使用反轉錄聚合酶連鎖反應 (RT-PCR)、西方墨點法(western blot) 檢測褪黑素的受體表現,還有齒原細胞和成骨細胞分化相關之基因跟蛋白的表現。也透過西方墨點法和免疫螢光染色 (immunofluorescent) 染色檢測信號分子的磷酸化。並使用 phalloidin 染色檢測肌動蛋白絲。實驗結果: 在人類牙根尖細胞中,有表現褪黑素受體MT1、MT2、MT3和RORα。加入褪黑素處理會增強成骨相關的轉錄因子Runx2和SP7的表現。對於跟細胞移動有關的作用,我們發現褪黑素會促進cofilin的磷酸化並刺激肌動蛋白絲聚合。但褪黑素並沒有改變cofilin-1本身的基因和蛋白表現。而H7會抑制褪黑素誘導的cofilin磷酸化,H89則反而更刺激磷酸化cofilin的表現。結論:這是首次檢測出人類牙根尖細胞中有表現褪黑素受體的研究。因為褪黑素對牙根尖細胞的影響是複雜的,並且可能會根據給藥物的劑量和作用時間而不同。值得進一步深入研究褪黑素的相關生物功能,這將會對未來的臨床治療很有幫助,無論是在再生牙髓病學領域或是骨頭再生跟重建的領域。 | zh_TW |
dc.description.abstract | Aim: Melatonin owns multiple biological functions in various tissues, and becomes widely used in multiple areas including stem cell biology and tissue engineering. Human apical papilla cells have been reported to show characteristics of stem cells and are known as stem cells from apical papilla (SCAP). The purpose of this study is to investigate the effects of melatonin on apical papilla cells and the role of PKA and PKC signaling pathways. Materials and methods: Primary-cultured human apical papilla cells were treated with different concentrations of melatonin and also with/without H89 (an inhibitor of PKA) or H7 (an inhibitor of PKC) pretreatment. Microscope observed cell morphology. Cell proliferation was measured by MTT assay. The expression of melatonin receptors, odontogenic/osteogenic transcription factors, differentiation markers, and actin depolymerizing factors were examined by reverse transcription polymerase chain reaction (RT-PCR) and western blot. Phosphorylation of signaling molecules was examined by western blot and immunofluorescent (IF) staining. Actin filaments were detected by phalloidin staining. Results: In human apical papilla cells, the expression of melatonin receptors, MT1, MT2, MT3/NQO2, and RORα were found. Treatment with melatonin (10~500 μg/ml) enhanced the expression of osteogenic transcription factors, Runx2 and SP7. In cell migration and morphogenesis, melatonin increased the phosphorylation of cofilin and stimulated actin filament polymerization. However, the mRNA and protein levels of cofilin1 were not changed by melatonin treatment. The enhancement of cofilin phosphorylation which induced by melatonin was repressed by H7, while H89 even stimulated the expression of p-cofilin. Conclusion: This is the first study to detect the expression of melatonin receptors in human apical papilla cells. However, the effect of melatonin on apical papilla cells is complicated and might be divergent depending on the dose and duration of treatment. Further investigating the biological activity of melatonin would be useful for clinical therapies in the future, including regenerative endodontics and bone regeneration. | en |
dc.description.provenance | Made available in DSpace on 2021-07-11T15:28:22Z (GMT). No. of bitstreams: 1 ntu-107-R04422005-1.pdf: 3735772 bytes, checksum: 944af26fd42ba16d49b1bcec85ee48df (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | 中文摘要 i
Abstract iii Table of Content vi List of Figures x List of Tables xiii Abbreviations List xiv Chapter I Literature Review 1 1.1 Regenerative endodontics 1 1.2 Stem cells from apical papilla 4 1.3 Signaling molecules and scaffold 6 1.4 Treatment procedures for regenerative endodontics 7 1.5 Melatonin 8 1.5.1 Mechanisms of melatonin action 9 1.5.2 Melatonin in tissue engineering and regenerative medicine 11 1.6 Odontogenic/osteogenic transcription factors 12 1.7 Odontogenic/osteogenic differentiation markers 12 1.8 Chemotaxis and cell migration 14 Chapter II The Purposes and Hypothesis of This Study 16 Chapter III Materials and Methods 17 3.1 The culture of human apical papilla cells 17 3.2 Observation of cell morphology 17 3.3 MTT assay 18 3.4 RNA extraction and reverse transcription-PCR 19 3.4.1 Isolation of total RNA 19 3.4.2 RNA quantitation 20 3.4.3 Reverse transcription (RT) 20 3.4.4 Polymerase chain reaction (PCR) 21 3.4.5 Gel electrophoresis 21 3.5 Western blot 22 3.5.1 Protein extraction 22 3.5.2 Protein quantification 22 3.5.3 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) 23 3.5.4 Transfer gel 23 3.5.5 Blocking and antibody hybridization 24 3.5.6 Chemiluminescence photography 25 3.6 Immunofluorescence (IF) staining 25 3.7 Phalloidin staining 26 3.8 Statistical analysis 27 Chapter IV Results 28 4.1 Expression of melatonin receptors 28 4.2 Cell morphological observation 28 4.3 Effects of melatonin on cell viability of apical papilla cells - MTT assay 28 4.4 Effect of melatonin on the expression of transcription factors 29 4.5 Effect of melatonin on the expression of odontogenic/osteogenic differentiation markers 29 4.6 Effect of melatonin on the regulation of cofilin 30 4.7 Effect of melatonin on the expression of N-cadherin 30 4.8 Effect of H89 on the melatonin regulation 30 4.9 Effect of H7 on the melatonin regulation 31 4.10 Effect of melatonin on the actin filament formation 31 4.11 Effect of melatonin on the expression of p-CREB 31 Chapter V Discussion 32 5.1 Tissue distribution of melatonin receptors 32 5.2 Effect of melatonin in odontogenesis and osteogenesis 33 5.3 Effect of melatonin on actin filaments 35 5.4 Potential signaling pathways involved in the biological activity of melatonin on human apical papilla cells 37 Chapter VI Conclusions 39 References 40 Figures 47 Tables 65 | |
dc.language.iso | en | |
dc.title | 褪黑素對於牙根尖細胞生物功能之影響 | zh_TW |
dc.title | Effect of Melatonin on the Biological Activity of Human Apical Papilla Cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 謝易修,楊順發,張美姬,林巧雯 | |
dc.subject.keyword | 再生牙髓病學,牙根尖細胞,根尖乳突幹細胞,褪黑素,褪黑素受體, | zh_TW |
dc.subject.keyword | regenerative endodontics,apical papilla cells,stem cells from apical papilla,melatonin,melatonin receptors, | en |
dc.relation.page | 69 | |
dc.identifier.doi | 10.6342/NTU201803949 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-08-19 | |
dc.contributor.author-college | 醫學院 | 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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