骨生成及免疫反應在骨母細胞與蝕骨前驅細胞共培養於矽酸鈣骨水泥上的評估

Osteogenic and immunogenesis potentials of co-cultured osteoblast cells and pre-osteoclast cells on calcium silicate cement

賴威勻(Wei-Yun Lai)

指導教授 ： 黃翠賢

中山醫學大學/口腔醫學院/牙醫學系碩士班/碩士(2013年)

摘要

MTA為近20年根管治療領域的重要材料，利用其優越的物化性質和生物相容性，於臨床上可進行破洞修補、根尖逆充填和各種覆髓相關的治療，本實驗室過去亦開發出新型的矽酸鈣水泥(CS cement) ，且與MTA具有相似的組成及性質，而當材料在臨床應用時常會與齒槽骨或是牙周組織有直接的接觸，因此材料與骨細胞接觸後的影響和關係是需要被深入了解的。本實驗目的在於探討骨母細胞與蝕骨前驅細胞共培養於材料表面時的骨生成反應和免疫反應。本實驗利用細胞適應性測試、電子顯微鏡、離子濃度分析、細胞週期分析以及西方墨點法等方法，分析MTA和CS對於細胞活性與細胞發炎反應的表現差異，並且以組織培養皿作為對照。結果發現CS對於骨母細胞都具有促進生長的效果，但CS卻會抑制蝕骨前驅細胞的生長，且在統計學上與TCP組相比有顯著差異。而MTA對於骨母細胞有顯著的促進生長效果，對蝕骨細胞則無顯著差異。電子顯微鏡下則顯示骨母細胞在MTA和CS表面都表現出良好的貼附，而蝕骨細胞在CS的表面則貼附不佳。在發炎因子和骨生成相關因子的實驗則顯示，CS能夠抑制發炎反應(TNF-α、iNOS、COX-2)(P<0.05)，且藉由刺激骨母細胞的生長來提高OPG的分泌和RANKL競爭RANK達到抑制蝕骨細胞的生長。

關鍵字

三氧礦化合物 ； 矽酸鈣水泥 ； 骨母細胞 ； 蝕骨細胞 ； 共同培養 ； 細胞分化

並列摘要

It is acknowledged that MTA has been one of the most significant developments in the endodontic field during the past 20 years. As MTA is characterized by superior physical, chemical properties along with biocompatibility, it is mainly used for perforation repair, root-end filling, and pulp capping related therapy. Our laboratory has also developed a novel calcium silicate cement (CS cement) with a similar composition and nature to the MTA. When in clinical application, these materials often directly contact with periodontal tissue and alveolar bone. Hence, the influence and relationship between the material and bone cell require an in-depth understanding. In this sense, the purpose of this study aims to investigate Osteogenic and immunogenesis potential of co-cultured osteoblast cells and pre-osteoclast cells on calcium silicate cement.
This study will employ cell compatibility test, cell cycle test, SEM, ion releasing test and western blotting to compare cell proliferation and inflammation between CS and MTA. Tissue culture plate was used as control. Results indicate that CS contributes greatly to stimulation for proliferation of osteoblast, while CS has an inhibiting effect on pre-osteoclast cells growth by showing statistical significance between CS and TCP(P<0.05). MTA also helps stimulate the proliferation of osteoblasts but no significant effect on pre-osteoclasts. SEM images show that osteoblast cells perform great cell adhesion on the surface of CS and MTA, while pre-osteoclast cells only adhere to the surface of MTA. In the experiments regarding inflammatory factors and osteogenic related facors, results reveal that CS can inhibit inflammation (TNF-α、iNOS、COX-2) by showing significant difference between CS and TCP (P<0.05), and restrain pre-osteoclast differentiation by stimulating osteoblast proliferation to increase OPG secretion which is competitive to RANKL for RANK.

並列關鍵字

MTA ； CS cement ； osteoblast ； pre-osteoclast ； co-culture ； cell differentiation

參考文獻

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