Analyses of Host-Biofilm Interactions: A Novel Polymicrobial Model

宿主-生物膜相互作用的分析：一种新型的多微生物模型

• 批准号: 7387045
• 负责人: Karen F. Novak
• 金额: $ 21.98万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7387045
• 关键词: Actinobacteria class; Actinomyces; Address; Adherent Culture; Antibiotic Resistance; Area; Bacteria; Biological; Biological Markers; Biological Models; Biological Process; Body Surface; Cell Culture System; Cell model; Cells; Characteristics; Chronic; Clinical; Complex; Confocal Microscopy; Contact Lenses; Coupled; DNA; Data; Dental Plaque; Deposition; Depth; Development; Disease; Disinfection; Disorder by Site; Documentation; Ecology; End Point; Epithelial; Epithelial Cells; Evaluation; Extended-Wear Contact Lenses; Fibroblasts; Fusobacteria; Fusobacterium; Gases; Genes; Gingiva; Gingivitis; Glycoproteins; Gram-Negative Anaerobic Bacteria; Grant; Harvest; Health Transition; Human; Immune; Immune response; Immunologics; In Situ; In Vitro; Individual; Infection; Inflammatory; Inflammatory Response; Knowledge; Lead; Lesion; Literature; Lymphocyte; Metabolic; Microbial Biofilms; Modeling; Natural Immunity; Nature; Numbers; Nutrient; Oral; Oral cavity; Pattern; Periodontal Diseases; Periodontitis; Polymerase Chain Reaction; Polymers; Principal Investigator; Process; Proteins; Reproducibility; Research; Research Design; Seminal; Silicones; Site; Standards of Weights and Measures; Stimulus; Streptococcus; Structure; Suspension substance; Suspensions; System; Testing; Time; Tissues; Tooth eruption; Tooth structure; Variant; Veillonella; alveolar bone; base; cell type; commensal microbes; copolymer; in vitro Model; innovation; lens; loss of function; macrophage; microbial; microorganism; novel; novel strategies; oral bacteria; oral biofilm; oral streptococci; pathogen; programs; quorum sensing; response; size; soft tissue; subgingival biofilm; theories; tooth surface

DESCRIPTION (provided by applicant): Periodontitis represents the most broadly distributed polymicrobial disease of mankind. The tissue destructive processes that lead to loss of function of periodontal soft tissues and alveolar bone result from a chronic polymicrobial biofilm activation of host inflammatory, innate immune, and adaptive immune responses in the local tissues. Development of the polymicrobial biofilm is a sequential process that begins with deposition of host and bacterial glycoproteins onto the clean tooth surface. Initial colonization by oral microorganisms occurs next, with oral streptococci being the primary early colonizers. This initial colonization by streptococci is followed by increasing proportions of Actinomyces, Veillonella, and Fusobacterium spp. (middle colonizers) with subsequent colonization predominated by Gram-negative, anaerobic bacteria (late colonizers). This shift from a predominance of Gram-positive, facultative microorganisms to a microbiota dominated by Gram- negative, anaerobic species is manifest clinically by inflammatory changes in the periodontal tissues as the host responds to this microbial challenge. It has been proposed that this multicellular bacterial-host interaction possesses unique features compared to planktonic bacterial infectious challenges; however, there is little objective evidence to support this contention. This R21 exploratory/development grant will specifically focus on developing this evidence using a novel polymicrobial biofilm model. The General Hypotheses to be tested are: (1) Oral bacteria in biofilms will elicit different patterns of host responses from gingival epithelial cells; and (2) Host cells will respond differently to bacterial species present as monospecies versus multispecies biofilms. We have developed a novel in vitro biofilm-host cell model system using a rigid, gas permeable contact lens material. In Specific Aim 1 we will determine the characteristics of single species and multispecies biofilms created on this material. The biofilms developed in this model system will then be used in Specific Aim 2 to challenge oral epithelial cells, allowing us to characterize the patterns of responses elicited in these host cells by the bacterial challenge. In accomplishing these aims, we will have developed a novel approach for evaluating host response patterns to planktonic, single species biofilm and polymicrobial biofilm challenges. The results obtained will enable us to provide seminal data to enable a further exploration of host responses to biofilms of various compositions and complexities. The bacteria found in dental plaque biofilms interact with tissues supporting the teeth to initiate an inflammatory response. The clinical diseases associated with this inflammatory response are gingivitis and/or periodontitis. However, the exact host responses elicited by this interaction have not been determined. We are developing an in vitro model of this interaction to gain a better understanding of the pathologenesis of gingivitis and periodontitis. This understanding is necessary to develop and evaluate new strategies for controlling the destructive features of these chronic inflammatory diseases.

描述（申请人提供）：牙周炎是人类分布最广泛的多种微生物疾病。导致牙周软组织和牙槽骨功能丧失的组织破坏性过程是由局部组织中宿主炎症、先天免疫和适应性免疫反应的慢性多微生物生物膜激活引起的。多微生物生物膜的形成是一个连续的过程，首先是宿主和细菌糖蛋白沉积到清洁的牙齿表面。口腔微生物的最初定植发生在接下来，口腔链球菌是主要的早期定植者。链球菌的最初定殖随后是放线菌、韦荣氏球菌和梭杆菌属的比例增加。（中期定殖者），随后的定殖由革兰氏阴性厌氧菌（晚期定殖者）主导。从占优势的革兰氏阳性兼性微生物到由革兰氏阴性厌氧物种占优势的微生物群的这种转变在临床上通过牙周组织中的炎性变化来表现，因为宿主响应于这种微生物挑战。有人提出，这种多细胞细菌-宿主相互作用具有独特的功能相比，嗜热菌感染的挑战，但是，很少有客观的证据来支持这一论点。这项R21探索/开发资助将特别侧重于使用新型多微生物生物膜模型开发这一证据。待检验的一般假设是：（1）生物膜中的口腔细菌将引起牙龈上皮细胞的不同宿主反应模式;（2）宿主细胞将对作为单种生物膜与多种生物膜存在的细菌物种作出不同的反应。我们已经开发了一种新的体外生物膜宿主细胞模型系统，使用刚性，透气接触透镜材料。在具体目标1中，我们将确定在这种材料上产生的单物种和多物种生物膜的特性。然后，在该模型系统中开发的生物膜将用于特定目标2以挑战口腔上皮细胞，使我们能够表征细菌挑战在这些宿主细胞中引起的反应模式。在实现这些目标，我们将开发一种新的方法来评估宿主的反应模式，以适应性，单一物种的生物膜和多微生物生物膜的挑战。所获得的结果将使我们能够提供开创性的数据，使主机响应各种组合物和复杂性的生物膜的进一步探索。在牙菌斑生物膜中发现的细菌与支持牙齿的组织相互作用，引发炎症反应。与这种炎症反应相关的临床疾病是牙龈炎和/或牙周炎。然而，这种相互作用引起的确切宿主反应尚未确定。我们正在开发一种这种相互作用的体外模型，以更好地了解牙龈炎和牙周炎的病理发生。这种理解对于开发和评估控制这些慢性炎症性疾病的破坏性特征的新策略是必要的。