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標題: | 不同根管封填劑和氧化鋅顆粒對MG-63類骨細胞之細胞週期及活性氧產生的影響及其機制 The effect and mechanism of root canal sealers and Zinc oxide particles on cell cycle control and reactive oxygen species production of MG-63 osteoblast-like cells |
作者: | Chia-Mei Tang 唐佳鋂 |
指導教授: | 鄭景暉 |
關鍵字: | MG-63類骨細胞,氧化鋅,G2 / M期停滯,第一型血紅素氧化?,氧化鋅丁香油酚根管充填劑, MG-63 osteoblast-like cells,Zinc oxide-eugenol-based root canal,Zinc oxide,cell cycle G2/M arrest,Hemo oxygenase-1, |
出版年 : | 2017 |
學位: | 碩士 |
摘要: | 實驗目的: 氧化鋅是根管封填劑的主要成分之一。本篇研究的目的主要是在比較不同劑量之市售氧化鋅丁香油酚基底的根管充填劑粉末與微米及奈米等級的氧化鋅顆粒在MG-63類骨細胞中對細胞週期和活性氧產生的影響。 實驗方法: 我們先在掃描電子顯微鏡下觀察藥物粉末的外觀。並將MG-63類骨細胞暴露在不同濃度(0-100μg/ ml)的市售氧化鋅丁香油酚基底根管充填劑 (Roth,canals),微米氧化鋅以及奈米氧化鋅24小時。於光學顯微鏡下觀察加藥24小時後MG-63細胞形態變化。並以反轉錄聚合酶連鎖反應(RT-PCR)西方墨點法(Western Blot)和免疫螢光法 (Immunofluorescence) 探討藥物對MG-63類骨細胞細胞週期,抗氧化反應和第一型膠原蛋白表現的影響。 實驗結果: 市售氧化鋅丁香油酚基底根管充填劑 (Roth®、canals®),微米氧化鋅以及奈米氧化鋅在抑制細胞增殖,細胞週期相關酵素以及抗氧化酵素的表現均類似。在光學顯微鏡下發現,當藥物濃度增加時,細胞數量有顯著的下降。而在反轉錄聚合酶連鎖反應(RT-PCR)西方墨點法(Western Blot)和免疫螢光法 (Immunofluorescence)的檢測中,我們發現cdc25c、cdc2、cycin b1表現量下降而GADD45α 表現量會增加,表示細胞週期停滯在G2 / M期。第一型血紅素氧化酶(HO-1)的表現量也隨著試劑濃度升高而增加。然而,p-Nrf2的表現量卻是減少,這可能表示HO-1的表現並不經由Nrf2途徑的調節。 p-p38和p-ERK的升高和p-Akt的下降也顯示她們在細胞功能的調節扮演重要的角色。 結論: 市售氧化鋅丁香油酚基底根管充填劑 (Roth®、canals®),微米氧化鋅以及奈米氧化鋅對於抑制細胞增殖,細胞週期相關酵素以及抗氧化酵素的表現均類似,它們使MG-63類骨細胞出現細胞週期停滯在G2 / M期和HO-1的表現量增加,其可能與p-p38和p-ERK路徑有關。 Aim: Zinc oxide is one of the main ingredient of root canal sealer. The purpose of the study focus on the effect of commercial ZOE root canal sealer powder and different size of Zinc oxide particles in the cell cycle and Reactive Oxygen Species production of MG63 osteoblast-like-cell. Materials and methods: MG63 osteoblast-like-cell were treated with Canals, Roth, micro-ZnO and nano-ZnO in different concentration (10-100 µg/ml) for 24 hours. Canals®, Roth®, micro-ZnO and nano-ZnO were observed under scanning electron microscope. The MG-63 cell morphology was observed under optical microscope. Reverse Transcription Polymerase Chain Reaction) (RT-PCR), Western Blot and Immunofluorescence were used to detect the effect of reagents on cell-cycle regulatory, antioxidant responses and collagen-1 expression. Results: All reagents inhibited cell proliferation in a dose-dependent manner. The decrease of cdc25c, cdc2, cycin b1 and the increase of GADD45α expression suggest the cell cycle arrest at G2/M phase after treatment. Antioxidant enzyme: heme oxygenase-1(HO-1) also expression as the reagents concentration rise. However, the p-Nrf2 expression is decrease which may suggest the activation of HO-1 may not regulated through Nrf2 pathway. The elevation of p-p38 and p-ERK and decline of p-Akt may also play a role in regulation cell functions. Conclusions: Canals®, Roth®, micro-ZnO and nano-ZnO have the similarly effect on inhibited cell proliferation as the concentration rise. They also induced cell cycle arrest at G2/M phase and HO-1 expression which may have relation with p-p38 and p-ERK pathway. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/20650 |
DOI: | 10.6342/NTU201702360 |
全文授權: | 未授權 |
顯示於系所單位: | 臨床牙醫學研究所 |
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