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三次元立体培養ヒト歯根膜線維芽細胞を用いた最適矯正力に関する検討

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Please use this identifier to cite or link to this item:http://doi.org/10.14943/doctoral.k11248
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Title: 三次元立体培養ヒト歯根膜線維芽細胞を用いた最適矯正力に関する検討
Other Titles: A study on optimum orthodontic force using human periodontal ligament cells on 3D culture system
Authors: 羽二生, 芽里 Browse this author
Keywords: 三次元立体培養ヒト歯根膜線維芽細胞
メカニカルストレス
サイ トカイン
Issue Date: 25-Mar-2014
Abstract: Osteoclast differentiation on the pressure side with the tooth movement mechanism by orthodontic forces is believed to be controlled by cytokines that periodontal ligament fibroblasts secrete; in experiments thus far, there have been reports using the methods of adding mechanical irritation directly to the cells. Therefore, in this study, We developed a three-dimensional culture system of periodontal ligament fibroblasts which assumes the periodontal ligament of in vivo and mechanical compression was applied. The aim of this study is to clarify the relationship of the compression amount and the changes of the cytokine gene expression levels that are related to the osteoclast differentiation. We cultured human periodontal ligament fibroblasts from teeth that were extracted for the purpose of orthodontic treatment. After a one week culture in which the culture cells of passage 5 were packed in a φ5×3 mm collagen sponge, the collagen sponges were compressed by1/3, 1/4. We searched for changes in the cytokine gene expression level in O (the control) after 1, 3, 6, 12, and 24 hours in real-time PCR. Although the packed state of the cells when compressed by 1/3 and 1/4 was similar in comparison to the control. The mRNA expression of cytokines TNF-α, FGF-2, IL-1β and VEGF related to osteoclast differentiation increased significantly more than the control at each point of 1/3 and 1/4 compression. The mRNA expression level of OPG was decreased without 24 hours after the starts of 1/3 compression. Overall, the relative a mounts of the mRNA expression were similar to the expression tendencies between the time of 1/3 compression and the time of 1/4 compression, but there was a higher tendency at the time of 1/4 compression. From these results, it has been suggested that when mechanical compression is added to the periodontal ligament fibroblasts in an experimental system that is assumed as in vivo, the cells produce cytokines and may play a part in bone remodeling. In addition, we believe it is effective to move teeth in or thodontic treatment when the tooth was pushed by a distance of a quarter of the width of the periodontal ligament.
矯正力による歯の移動機構で圧迫側における破骨細胞分化は,歯根膜線維芽細胞が分泌するサイトカインにより制御されると考えられており,今までの実験では,直接細胞に機械的刺激を加えた報告が多かった.そこで,本研究ではinvivoに近い歯根膜を想定し,三次元立体培養歯根膜線維芽細胞を作製して機械的圧縮を加え,産生される破骨細胞分化関連サイトカイン遺伝子発現量の変化と圧縮量の関係を明らかにすることを目的とした.矯正治療時便宜抜去された歯からヒト歯根膜線維芽細胞を培養し,5継代目の培養細胞をφ5º3mmのコラーゲンスポンジに充填し1週間培養後,コラーゲンスポンジを1/3,1/4圧縮した.0(コントロール),1,3,6,12,24時間後にサイトカイン遺伝子発現量の変化をReal time PCRにて検索した.1/3, 1/4圧縮時の細胞の充填状態は,コントロールと比べ同程度であった.破骨細胞分化関連サイトカインであるTNF-α,FGF2,IL-1β,VEGFのmRNA発現は,1/3, 1/4圧縮すべての時点でコントロールより有意に増加し,OPGのmRNA発現量は,1/3圧縮24時間後の条件以外有意に減少していた.1/3圧縮24時間後のOPGのmRNA発現量は,コントロールと同程度であった.また,総じて,mRNA発現の相対量は1/3圧縮時と1/4圧縮時の間で発現傾向は類似していたが,1/4圧縮時の方が多い傾向であった.これらの結果からin vivoに近い実験系で歯根膜線維芽細胞に機械的圧縮を加えたところ,細胞がサイトカインを産生し,骨リモデリングの一端を担っている可能性が示唆された.また,矯正治療において歯根膜幅の1/4の移動を行って歯を移動させることが効率的であると考えられる.
Conffering University: 北海道大学
Degree Report Number: 甲第11248号
Degree Level: 博士
Degree Discipline: 歯学
Examination Committee Members: (主査) 教授 飯田 順一郎, 教授 鈴木 邦明, 教授 田村 正人
Degree Affiliation: 歯学研究科(口腔医学専攻)
Type: theses (doctoral)
URI: http://hdl.handle.net/2115/55748
Appears in Collections:学位論文 (Theses) > 博士 (歯学)
課程博士 (Doctorate by way of Advanced Course) > 歯学院(Graduate School of Dental Medicine)

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