Biofilm Architecture of Subgingival Plaque

龈下菌斑的生物膜结构

• 批准号: 7472252
• 负责人: Renate Lux
• 金额: $ 7.9万
• 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 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7472252
• 关键词: Actinobacillus actinomycetemcomitans; Address; Adult; Affect; Architecture; Bacteria; Bacterial Infections; Biological Models; Body Fluids; Cell surface; Cells; Chronic; Communities; Complex; Computer software; Condition; Culture Techniques; Data Analyses; Detection; Development; Diagnostic; Diagnostic tests; Disease; Electron Microscopy; Environment; Equipment and supply inventories; Fluorescence; Fluorescence Microscopy; Fluorescent in Situ Hybridization; Forsythia; Fusobacteria; Fusobacterium; Genetic; Heart Diseases; Image; Image Analysis; Imagery; Immune system; In Situ Hybridization; In Vitro; Individual; Infection; Integration Host Factors; Label; Laboratories; Lead; Maps; Medical; Methods; Microbial Biofilms; Microscopic; Mission; Molecular; Monitor; Nature; Numbers; 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; 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）：全球大量的成年人群受牙周病的影响。这些是牙齿缺失组织的慢性生物膜感染也与全身性疾病有关。即使可以提供有关口腔生物膜的口腔菌群和一般结构信息的大量清单，但在很大程度上缺少了对物种分布及其在生物膜中的相互作用的详细分析。该项目的长期目的是建立荧光原位杂交（FISH），以详细检查重要的亚生物生物膜特征，包括物种存在和丰度，本地化模式，生物膜特异性形型，ASET培养有机体的作用以及对生物生物结构形成的细菌作用。选择鱼类作为工具，因为这种多功能技术可以在所有分类水平上独立培养微生物的培养物检测，并在不受干扰的样品中可视化它们的分布。此外，特定微生物在建筑生物膜方面及其相互作用的作用将在定量水平上进行检查。根据NIDCR的任务，这将为开发新的诊断和医疗靶标的重要信息，以治疗牙周疾病，由于感染的持续性，这些牙周疾病仍然难以治愈。在此特定应用中，我们建议使用鱼类来确定生物膜群落中选定推定的牙周病原体的比例，空间分布和共同定位模式。该方法将与基于载体的体内模型系统结合使用，该系统允许自然种植的牙周生物膜采样。使用特定的鱼类探针，将检查这些生物膜是否存在可疑的口腔病原体（例如口腔treponemes，fusobacteriumes，fusobacteriums ssp。，牙卟啉单胞菌，Tannerella Forsythia，Tannerella Forsythia，filifactor alocis，tm7 Complect）。对整个载体和切片载体的显微镜检查将对它们的丰度，空间分布和共定位（微生物之间相互作用的指示）进行定性评估。为了对获得的数据进行Quanitative分析，将采用针对被标记为样品的鱼类优化的新型数字图像分析程序DAIME。 这项研究将验证当前的牙周疾病发展思想，并提供有关涉及细菌的详细信息，包括在实验室试管培养物中无法研究的细菌。知道哪些细菌在牙周炎中生物膜群落积聚中起着核心作用，可以鉴定出新的标记生物以进行诊断测试。这可能会导致治疗方法与不幸免的有益细菌的现有疗法的“杀死”方法相比，只有关键的有害生物被消除。此外，可以轻松监控治疗和治疗结果的失败。 项目叙述：牙齿支持组织（牙龈和骨骼）的慢性细菌感染，牙周疾病会影响大部分成年人。它是牙齿脱落的主要原因，可能与所谓的全身状况（例如心脏病）有关。牙周疾病很难永久治愈。重复治疗对于保留这些感染的原因是必要的，因为它们的原因，高度结构化的细菌群落和牙周口袋内的未知细菌很难完全去除。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，这些结构将其他细菌固定在适当的位置，最丰富的细菌以及哪种类型的细菌可能相互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。