Impact of Oral bacterial interactions on periodontitis

口腔细菌相互作用对牙周炎的影响

• 批准号: 6606340
• 负责人: ERIC C REYNOLDS
• 金额: $ 16.2万
• 依托单位: UNIVERSITY OF MELBOURNE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6606340
• 关键词: Bacteroides gingivalis; Fusobacterium nucleatum; Treponema; bacteria infection mechanism; bacterial antigens; bacterial proteins; bactericidal immunity; bone disorder; disease /disorder model; host organism interaction; infection; laboratory mouse; mandible /maxilla; mass spectrometry; membrane proteins; microorganism culture; oral bacteria; periodontitis; two dimensional gel electrophoresis; western blottings

DESCRIPTION (provided by applicant): Periodontal diseases are bacterial-associated inflammatory diseases of the supporting tissues of the teeth, and one of the more aggressive forms of chronic periodontitis is characterized by the destruction of the tooth's supporting structures. Chronic periodontitis has a large economic burden and is a major public health problem. Three bacterial species (Porphyromonas gingivalis, Treponema denticola, Bacteroides forsythus) have been shown to be closely associated with chronic periodontitis in humans, and a commensal oral bacterium Fusobacterium nucleatum has been implicated in aiding the colonization of the periodontopathic bacteria. P. gingivalis, T. denticola, B. forsythus and F. nucleatum exist as part of a high density mixed species (polymicrobial) biofilm in the gingival crevice of humans. Recent evidence using rodent abdominal lesion models has indicated that co-infection of B. forsythus or T. denticola with P. gingivalis enhances lesion development. This synergistic interaction that enhances lesion severity, and potentially virulence of the bacteria, has not been studied in an animal periodontitis model. Further, outer membrane proteins that are altered during this interaction also have not been studied. The broad objective of this application is to determine the impact of interactions between P. gingivalis, T. denticola, B. forsythus and F. nucleatum on the level of bone loss and host antibody response in the murine periodontitis model and to characterize the major antigenic outer membrane proteins of these bacteria using 2D PAGE and mass spectrometric methods. Using these techniques, we have already identified over thirty P. gingivalis outer membrane proteins. We have also established a murine periodontitis model based on oral infection of P. gingivalis. These methodologies will be applied to determine the impact of bacterial interactions on the expression of major antigenic outer membrane proteins and on colonization, bone loss and host immune response. The significance of this work is that it will increase our knowledge of the role of bacterial interaction on polymicrobial virulence in the murine periodontitis model and may have a broader significance in the study of human mucosal pathogens. The specific aims of the application are: 1) To determine the extent of alveolar bone loss and level of bacterial colonization in the murine periodontitis model when co-inocula of P. gingivalis, T. denticola, B. forsythus and/or F. nucleatum grown as monocultures are used to orally infect mice compared with mono-inoculation of each species. 2) To determine the extent of alveolar bone loss and bacterial colonization in the murine periodontitis model when polymicrobial cultures of P. gingivalis, T. denticola, B. forsythus and/or F. nucleatum are used to orally infect mice compared with mono- and co-inocula of the bacteria grown as monocultures as in (1) above. 3) To determine the host antibody isotype response to each of the bacteria in the murine periodontitis model when co-inocula or polymicrobial cultures of P. gingivalis, T. denticola, B. forsythus and/or F. nucleatum are used to orally infect mice. 4) To identify the major outer membrane and antigenic proteins of P. gingivalis, T. denticola, B. forsythus and F. nucleatum grown in monoculture and as polymicrobial cultures using 2D-gel electrophoresis, mass spectrometry and Western blotting.

描述（由申请人提供）：牙周疾病是牙齿支撑组织的细菌相关炎症性疾病，而慢性牙周炎的一种更具侵略性的形式之一的特征是破坏了牙齿的支撑结构。 慢性牙周炎具有巨大的经济负担，是一个主要的公共卫生问题。 已经证明，三种细菌种类（斑岩菌，treponema牙齿牙皮植物，细菌牙齿，细菌剂）与人类的慢性牙周炎密切相关，共同口腔细菌核细菌已涉及拟宣布牙周病性细菌的共同化。 P. gingivalis，T。Denticola，B。Forsythus和F. nuteatum是人类牙龈缝隙中高密度混合物种（多数菌）生物膜的一部分。 使用啮齿动物腹部病变模型的最新证据表明，与牙龈疟原虫的Forsythus或T. denticola共同感染可增强病变的发育。 在动物牙周炎模型中尚未研究这种增强病变严重程度和潜在毒力的协同相互作用。 此外，在这种相互作用过程中改变的外膜蛋白也尚未研究。 该应用的广泛目的是确定牙龈疟原虫，牙齿状球芽孢杆菌，B。forsythus和F. nucleatum对鼠牙周炎模型中骨质流失和宿主抗体反应水平的影响，并使用2D页和质量谱法的这些细菌的主要抗原外膜外膜蛋白来表征这些主要的抗原外膜外膜蛋白。 使用这些技术，我们已经确定了三十多张牙龈牙周膜外膜蛋白。 我们还建立了基于口服牙龈疟原虫感染的鼠牙周炎模型。 这些方法将应用于确定细菌相互作用对主要抗原外膜蛋白表达以及定植，骨丢失和宿主免疫反应的影响。 这项工作的意义在于，它将增加我们对鼠牙周炎模型中细菌相互作用对多因素毒力的作用的了解，并且在人类粘膜病原体的研究中可能具有更广泛的意义。 该应用的具体目的是：1）确定鼠牙周炎模型中肺泡骨质流失和细菌定殖水平的程度，当牙龈疟原虫，T。Denticola，牙霉菌，B。Forsythus和/或F. forsythus和/或F. nucleatum种植，单次培养物与单单培养物相比，与每种物种相比。 2）当在鼠牙周炎模型中确定肺泡骨质流失和细菌定殖的程度，当时牙龈疟原虫的多菌血培养物，T。denticola，B。forsythus和/或F. nucleatum用于口服小鼠与单核和单培养物相比（1）等于（1），而不是生长的细菌。 3）确定鼠牙周炎模型中对每种细菌的宿主抗体同种型反应，当时牙龈疟原虫，T。denticola，B。Forsythus和/或F. nucletum用于口服感染小鼠时，牙齿牙周炎模型中的每种细菌。 4）鉴定牙龈疟原虫的主要外膜和抗原蛋白，T。denticola，B。Forsythus和F. nucleatum在单一培养物中生长的核和核，并使用2D-凝胶电泳，质谱和蛋白质印迹。