Guggenheim B, Giertsen E, Schüpbach P, Shapiro S
Institute for Oral Microbiology and General Immunology, Center for Dentistry, Oral Medicine, and Maxillofacial Surgery, University of Zürich, Switzerland.
J Dent Res. 2001 Jan;80(1):363-70. doi: 10.1177/00220345010800011201.
The study of biofilm structure and function mandates the use of model systems for which a host of environmental variables can be rigorously controlled. We describe a model of supragingival plaque containing Actinomyces naeslundii, Veillonella dispar, Fusobacterium nucleatum, Streptococcus sobrinus, and Streptococcus oralis wherein cells are cultivated anaerobically in a saliva-based medium on hydroxyapatite discs coated with a salivary pellicle, with material and pieces of apparatus common to all microbiology laboratories. After 0.5 hr, 16.5 hrs, 40.5 hrs, and 64.5 hrs, the composition of adherent biofilms was analyzed by culture techniques, live/dead fluorescence staining, and confocal laser scanning microscopy. Repeated independent trials demonstrated the repeatability of biofilm formation after 40.5 hrs and 64.5 hrs. Brief exposures of biofilms to chlorhexidine or Triclosan produced losses in viability similar to those observed in vivo. This biofilm model should prove very useful for pre-clinical testing of prospective anti-plaque agents at clinically relevant concentrations.
生物膜结构与功能的研究需要使用能够严格控制大量环境变量的模型系统。我们描述了一种包含内氏放线菌、差异韦荣球菌、具核梭杆菌、远缘链球菌和口腔链球菌的龈上菌斑模型，其中细胞在涂有唾液薄膜的羟基磷灰石圆盘上，于基于唾液的培养基中进行厌氧培养，使用的材料和仪器均为所有微生物实验室常用的。在0.5小时、16.5小时、40.5小时和64.5小时后，通过培养技术、活/死荧光染色和共聚焦激光扫描显微镜对附着生物膜的组成进行分析。重复的独立试验证明了在40.5小时和64.5小时后生物膜形成的可重复性。生物膜短暂暴露于洗必泰或三氯生后，活力损失与体内观察到的相似。这种生物膜模型对于在临床相关浓度下对潜在抗菌斑剂进行临床前测试应该非常有用。
