【摘要】：目的通過體外實驗建立根尖生物膜模型,觀察糞腸球菌根尖生物膜模型的形態(tài)及結(jié)構(gòu)。方法選取人新鮮拔除的單根離體牙24顆,隨機分為8組,每組3顆,浸泡于糞腸球菌ATCC 29212菌懸液中,分別于1、2、7 d建立根尖生物膜模型。使用掃描電子顯微鏡(SEM)觀察生物膜形成過程;使用碘化丙啶(PI)和刀豆球蛋白A異硫氰酸熒光素(ConA-FITC)對生物膜細菌及細胞外基質(zhì)進行雙染色,通過激光共聚焦掃描電子顯微鏡(CLSM)觀察其形態(tài)和結(jié)構(gòu);計算生物膜覆蓋率。結(jié)果隨時間的增加,根尖樣本表面細菌覆蓋面積增加,7 d組與1、2 d組間的差異存在統(tǒng)計學(xué)意義(P0.05),1 d與2 d組間的差異無統(tǒng)計學(xué)意義(P0.05)。7 d組形成生物膜結(jié)構(gòu),其樣本表面的覆蓋率為17.23%±1.52%,樣本表面有大量細菌附著,細菌被細胞外基質(zhì)包繞,并呈群落分布。生物膜中的細菌被PI染成紅色,細胞外基質(zhì)被ConA-FITC染成綠色,可見多層次的空間結(jié)構(gòu)以及親水性通道。結(jié)論糞腸球菌根尖生物膜于培養(yǎng)7 d時形成完整的生物膜結(jié)構(gòu),由細菌群落及其周圍大量的不定型基質(zhì)組成。
[Abstract]:Objective to establish the root tip biofilm model in vitro and observe the morphology and structure of Enterococcus faecalis root tip biofilm model. Methods A total of 24 isolated teeth were randomly divided into 8 groups, 3 in each group, soaked in ATCC 29212 suspension of Enterococcus faecalis. The model of biofilm was established at 1, 2 and 7 days, respectively. The process of biofilm formation was observed by scanning electron microscope (SEM). The biofilm bacteria and extracellular matrix were double stained with pyridine iodide (PI) and concanavalin A isothiocyanate (ConA-FITC). The morphology and structure of biofilm were observed by confocal scanning electron microscopy (CLSM). The biofilm coverage was calculated. Results with the increase of time, the bacterial coverage area of root tip samples increased, and the difference between 7 d group and 1 2 d group was statistically significant (P0.05). There was no significant difference between the 1 d and 2 d groups (P0.05). The biofilm structure was formed in the 7th day group. The coverage of the sample surface was 17.23% 鹵1.52%. A large number of bacteria were attached to the sample surface and the bacteria were surrounded by extracellular matrix. And distributed in community. The bacteria in the biofilm were stained red by PI, and the extracellular matrix was stained green by ConA-FITC. Conclusion the biofilm of Enterococcus faecalis root tip formed a complete biofilm structure after 7 days of culture and was composed of a large number of amorphous substrates around the bacterial community.
【作者單位】： 首都醫(yī)科大學(xué)附屬北京口腔醫(yī)院牙體牙髓科;
【基金】：國家自然科學(xué)基金面上項目資助項目(81170952) 首都衛(wèi)生發(fā)展科研專項基金資助項目(首發(fā)2011-2014-05)
【分類號】：R781.05

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本文編號：2339696