牙周菌斑生物膜体外模型建立.doc

牙周菌斑生物膜体外模型建立[摘要]目的观察血链球菌、牙龈卟啉单胞菌和具核梭杆菌在人工牙根面上随培养时间变化形成单菌种、双菌种和三菌种生物膜的能力，期望在体外建立龈下菌斑生物膜模型。方法培养血链球菌、牙龈卟啉单胞菌和具核梭杆菌试验菌株，制备胶原包被羟磷灰石的人工牙根面，在人工牙根面上培养形成试验菌株的单菌种、双菌种和多菌种生物膜，并用扫描电镜观察三种试验菌株分别在培养24、48和72 h后形成单菌种、双菌种和三菌种生物膜的情况。结果单独培养24 h的血链球菌在人工牙根面上形成的完整生物膜，具备三维立体结构；培养48和72 h后，细菌密度逐渐增大，生物膜更加成熟。单独培养48 h的牙龈卟啉单胞菌形成的完整生物膜，具备三维立体结构；培养72 h后，细菌密度有所降低。牙龈卟啉单胞菌和具核梭杆菌培养24 h形成的完整的双菌种生物膜，具备三维立体结构；培养48和72 h后已看不出完整的生物膜结构，两菌的数量大幅度降低。血链球菌、牙龈卟啉单胞菌和具核梭杆菌在培养24 h时形成的较完整的三菌种生物膜，初步具备三维立体结构；三菌中，血链球菌、具核梭杆菌所占比列远大于牙龈卟啉单胞菌；培养48 h后，三菌种生物膜更加成熟，牙龈卟啉单胞菌所占比例有所增加，此时的生物膜已具备三维立体结构；培养72 h后，三菌种仍保持较完整的生物膜结构，但数量均有所下降。结论血链球菌、牙龈卟啉单胞菌和具核梭杆菌在培养24 h时间点已基本形成了完整的单菌种、双菌种和三菌种生物膜结构，故在今后建立龈下菌斑生物膜模型时可选取24 h这一时间点。 [关键词]血链球菌；具核梭杆菌；牙龈卟啉单胞菌；生物膜；人工牙根面 [中图分类号]R 780.2[文献标志码]A[doi]10.3969/j.issn.1673-5749.2012.01.010 A pilot study of an model of periodontal biofilm in vitroLi Yongkai1, Duan Dingyu2, Zhao Lei2，3, Wu Yafei2，3, Xu Yi2, 3.（1. Dept. of Periodontics and Mucosa Disease, The Affiliated Hospital of Stomatology, Chongqing Medical University, Chongqing 401147, China; 2. State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, China; 3. Dept. of Periodontics, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China） [Abstract]ObjectiveThe ability of the single-species and multi-species biofilm formation on the artificial root surface by Streptococcus sanguis（S.sanguis）, Fusobacterium nucleatum（F.nucleatum）and Porphyromonas gingivalis（P.gingivalis）was observed in order to establish an in vitro subgingival plaque biofilm model. MethodsS.sanguis, F.nucleatum and P.gingivalis were cultured, and collagen-treated hydroxyapatite was prepared as the artificial root surface. The formation of the single-species, dual-species and multi-species biofilms of S.sanguis, F.nucleatum and P.gingivalis on the artificial root surface were detected at 24, 48 and 72 h by scanning electron microscopy. ResultsS.sanguis biofilm was formed on the artificial root surface at 24 h, and showed a three-dimens