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Role of coaggregation among periodontopathic bacteria in formation of periodontopathic biofilm

牙周病细菌共聚集在牙周病生物膜形成中的作用

基本信息

批准号：
16591837
负责人：
ISHIHARA Kazuyuki
金额：
$ 1.79万
依托单位：
Tokyo Dental College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

项目摘要

Objects : Periodontitis is caused by Gram-negative rods and spirochetes. Porphyromonas gingivalis is a major pathogen of chronic periodontal disease, This microorganism has a number of virulence factors, including fimbriae, lipopolysaccharides and proteases, and is often co-isolated with Tannerella forsythensis and Treponema denticola. The combination of these three microorganisms is known as the "red complex", and is thought to play an important role in the progression of chronic periodontitis. In this project, we investigated coaggregation and synergistic effects among these periodontopathic bacteria to clarify the underlying mechanisms of periodontopathic biofilm formation.Materials and Methods : Coaggregation between P.gingivalis ATCC 33277 and T.denticola ATCC 35405 was evaluated by absorbance at 660 nm in coaggregation buffer. To clarify which P.gingivalis ligands were involved in coaggregation, knockout mutants lacking gingipains and fimbriae were constructed. Synergistic effect … More s on biofilm formation were evaluated by using a two-compartment co-culture system incorporating a 0.4 m filter unit.Results and Discussion : Coaggregation reaction between P.gingivalis and T.denticola was decreased in the Mfa1 deficient mutant, kgp, RgpA and RgpB deficient mutant and Kgp, RgpA and HagA deficient mutant. These results indicated that gingipains and Mfa1 were involved in the coaggregation reaction. In particular, activity decreased significantly in the Kgp, RgpA and RgpB deficient strain and Kgp, RgpA and HagA deficient strain. In addition, coaggregation activity was observed in the Mfa1 FimA mutant. This suggests that the hemagglutinin/ahesion domains of gingipains and HagA play a key role in the coaggregation reaction. Using our two-compartment system, Fusobacterium nucleatum showed an approximately four-fold increase in biofilm formation in co-culture with P.gingivalis compared with that obtained with single culture alone. Taken together, these results suggest that the coaggregation reaction of P.gingivalis with other periodontopathogens is involved not only in the colonization of gingival crevicular biofilms, but also in the formation of periodontopathic biofilms. Less

目的：牙周炎是由革兰氏阴性杆菌和螺旋体引起的。牙龈卟啉单胞菌是慢性牙周病的主要病原菌，该菌具有菌毛、脂多糖和蛋白酶等多种毒力因子，常与福赛氏单胞菌和齿密螺旋体共分离。这三种微生物的组合被称为“红色复合体”，被认为在慢性牙周炎的进展中起着重要作用。材料和方法：用共聚集缓冲液中660nm处的吸光度来评价牙周炎假单胞菌ATCC 33277和齿假单胞菌ATCC 35405之间的共聚集性。为了弄清牙龈假单胞菌的哪些配体参与共聚集，构建了缺失牙龈痛和菌毛的基因敲除突变体。协同效应…结果和讨论：在Mfa1缺陷突变株、Kgp、RgpA和RgpB缺陷突变株以及Kgp、RgpA和HagA缺陷突变株中，牙龈假单胞菌与齿状念珠菌之间的共聚集反应降低。这些结果表明，牙周疼痛和MFA1参与了共聚集反应。尤其是Kgp、RgpA、RgpB缺陷株和Kgp、RgpA、HagA缺陷株，酶活性显著降低。此外，在Mfa1FimA突变体中观察到共聚集活性。这表明牙龈痛和HagA的血凝素/粘附区在共聚集反应中起着关键作用。使用我们的两室系统，核梭杆菌与牙龈假单胞菌共培养时，与单独培养相比，生物膜的形成大约增加了四倍。综上所述，这些结果表明，牙龈假单胞菌与其他牙周病原体的共聚反应不仅参与了牙周沟生物膜的定植，也参与了牙周病理性生物膜的形成。较少