Biofilm Architecture of Subgingival Plaque

龈下菌斑的生物膜结构

• 批准号: 7629051
• 负责人: Renate Lux
• 金额: $ 6.55万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7629051
• 关键词: Actinobacillus actinomycetemcomitans; Address; Adult; Affect; Architecture; Bacteria; Bacterial Infections; Biological Models; Body Fluids; Cell surface; Cells; Chronic; Communities; Complex; Computer software; Culture Techniques; Data Analyses; Detection; Development; Diagnostic; Diagnostic tests; Disease; Electron Microscopy; Environment; Equipment and supply inventories; Fluorescence Microscopy; Fluorescent in Situ Hybridization; Forsythia; Fusobacteria; Fusobacterium; Genetic; Heart Diseases; Image; Image Analysis; Imagery; Immune system; In Vitro; Individual; Infection; Integration Host Factors; Label; Laboratories; Lead; Maps; Medical; Methods; Microbial Biofilms; Microscopic; Mission; Molecular; Monitor; National Institute of Dental and Craniofacial Research; Nature; Oral; Organism; Patients; Pattern; Periodontal Diseases; Periodontal Pocket; Periodontitis; Play; Population; Porphyromonas gingivalis; Prevalence; Prevotella intermedia; Process; Qualitative Evaluations; Research; Role; Sampling; Selenomonas sputigena; Severity of illness; Site; Spatial Distribution; Structure; Systemic disease; Techniques; Testing; Therapeutic; Time; Tissues; Tooth Loss; Tooth structure; Treatment outcome; Treponema; Tube; Work; antimicrobial; base; bone; digital imaging; in vivo; in vivo Model; insight; killings; microbial; microbial community; microorganism; novel; novel diagnostics; novel marker; oral bacteria; oral biofilm; oral pathogen; pathogen; programs; subgingival biofilm; tool

DESCRIPTION (provided by applicant): A significant number of the adult population world-wide is affected by periodontal diseases. These chronic biofilm infections of the tooth-supporting tissues that are the major cause of tooth loss have also been associated with systemic diseases. Even though extensive inventory of the oral flora and general structural information about oral biofilms are available, detailed analysis of the species distribution and their interactions in biofilms causing periodontal disease is largely missing. The long-term objective of this project is to establish fluorescence in situ hybridisation (FISH) for the detailed examination of important subgingival biofilm features, including species presence and abundance, localization pattern, biofilm-specific morphotype, the role of as-yet-uncultured organisms and indentification of bacteria with pivotal role in biofilm structure formation. FISH is chosen as a tool since this versatile technique enables culture independent detection of microorganisms on all taxonomic levels and visualisation of their distribution in undisturbed samples. Furthermore, the role of specific microorganisms in architectoral biofilm aspects as well as their interaction will be examined on a quantitative level. In compliance with the mission of the NIDCR, this will provide important information for the development of new diagnostic and medical targets in the treatment of periodontal diseases which are still difficult to cure due to the persistent nature of the infection. In this specific application we propose to employ FISH to determine proportion, spatial distribution and co- localisation pattern of selected putative periodontal pathogens within the biofilm community. This method will be applied in combination with a carrier-based in vivo model system that allows sampling of naturally grown periodontal biofilms. These biofilms will be examined for the presence of suspected oral pathogens (e.g. oral treponemes, Fusobacterium ssp., Porphyromonas gingivalis, Tannerella forsythia, Filifactor alocis, TM7 complex) using specific FISH probes. Microscopic examination of whole and sectioned carriers will enable qualitative evaluation of their respective abundance, spatial distribution and co-localisation (indicative for interactions between microorganisms). To enable quanitative analysis of the data obtained, the novel digital image analysis program daime that was optimized for FISH labeled samples will be employed. This study will validate current ideas of periodontal disease development and provide detailed information about the bacteria involved including those that cannot be studied in laboratory test tube cultures. Knowing which bacteria play central roles in the build-up of the biofilm communities in periodontitis could identify new marker organisms for diagnostic tests. This could lead to treatments in which only key harmful organisms will be eliminated compared to the "kill all" approaches of existing therapies in which beneficial bacteria are not spared. Furthermore, failed treatments and treatments outcomes could be easily monitored. Project Narrative: Chronic bacterial infections of the tooth-supporting tissue (gum and bone), the periodontal diseases affect a significant portion of adults. It is the major cause for tooth loss and may be related to so-called systemic conditions such as heart disease. Periodontal diseases are difficult to cure permanently. Repetitive treatments are necessary to keep these infections in check since their cause, highly structured bacterial communities and yet unknown bacteria that reside within the periodontal pockets are difficult to remove completely. Our research will help to understand which type of bacteria are relevant for formation of these anchoring structures that keep other bacteria in place, who is most abundant and which types of bacteria may interact with each other to promote disease. If successful, this study will provide useful information for the development of new diagnostic and therapeutic tools.

描述（由申请人提供）：全世界有相当数量的成年人受到牙周病的影响。牙齿支持组织的这些慢性生物膜感染是牙齿脱落的主要原因，也与全身性疾病有关。尽管广泛的库存的口腔植物群和一般结构的信息，口腔生物膜，详细分析的物种分布和它们之间的相互作用，导致牙周病在很大程度上是缺失的。本项目的长期目标是建立荧光原位杂交（FISH），用于详细检查重要的龈下生物膜特征，包括物种存在和丰度、定位模式、生物膜特异性形态型、尚未培养的生物体的作用以及在生物膜结构形成中起关键作用的细菌的鉴定。选择FISH作为工具，因为这种多功能技术能够在所有分类学水平上独立检测微生物，并可视化其在未受干扰样品中的分布。此外，特定微生物在建筑生物膜方面的作用以及它们之间的相互作用将在定量水平上进行研究。根据NIDCR的使命，这将为开发治疗牙周病的新诊断和医疗目标提供重要信息，由于感染的持续性，牙周病仍然难以治愈。在该具体应用中，我们提出采用FISH来确定生物膜群落内选定的推定牙周病原体的比例、空间分布和共定位模式。该方法将与基于载体的体内模型系统结合使用，该系统允许对自然生长的牙周生物膜进行采样。将检查这些生物膜是否存在疑似口腔病原体（例如口腔密螺旋体、梭杆菌属，牙龈卟啉单胞菌、Tannerella gingivalis、Filifactoralocis、TM7复合物）。整个和切片载体的显微镜检查将能够对其各自的丰度、空间分布和共定位（指示微生物之间的相互作用）进行定性评价。为了能够对获得的数据进行定量分析，将采用针对FISH标记的样品进行优化的新型数字图像分析程序daime。 这项研究将验证目前牙周病发展的想法，并提供有关细菌的详细信息，包括那些不能在实验室试管培养研究。了解哪些细菌在牙周炎生物膜群落的建立中起着核心作用，可以为诊断测试确定新的标记生物。这可能导致只有关键的有害生物将被消除的治疗方法相比，“杀死所有”的方法，其中有益的细菌也不能幸免。此外，可以容易地监测失败的治疗和治疗结果。 项目叙述：牙齿支持组织（牙龈和骨骼）的慢性细菌感染，牙周病影响了相当一部分成年人。它是牙齿脱落的主要原因，可能与所谓的全身性疾病有关，如心脏病。牙周病很难根治。重复治疗是必要的，以保持这些感染的检查，因为他们的原因，高度结构化的细菌群落，但未知的细菌，居住在牙周袋是难以完全清除。我们的研究将有助于了解哪种类型的细菌与这些锚定结构的形成有关，这些锚定结构使其他细菌保持在适当的位置，谁是最丰富的，以及哪种类型的细菌可能相互作用以促进疾病。如果成功，这项研究将为开发新的诊断和治疗工具提供有用的信息。

项目成果

Co-localized or randomly distributed? Pair cross correlation of in vivo grown subgingival biofilm bacteria quantified by digital image analysis.

共定位还是随机分布？

• DOI: 10.1371/journal.pone.0037583
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Schillinger,Claudia;Petrich,Annett;Lux,Renate;Riep,Birgit;Kikhney,Judith;Friedmann,Anton;Wolinsky,LawrenceE;Göbel,UlfB;Daims,Holger;Moter,Annette
• 通讯作者: Moter,Annette

Molecular characterization of the microbial flora residing at the apical portion of infected root canals of human teeth.

• DOI: 10.1016/j.joen.2011.06.020
• 发表时间: 2011-10
• 期刊: Journal of endodontics
• 影响因子: 4.2
• 作者: Chugal N;Wang JK;Wang R;He X;Kang M;Li J;Zhou X;Shi W;Lux R
• 通讯作者: Lux R