PDF(Pubmed)
Abstract：
UNASSIGNED: Periodontal diseases are known to be associated with polymicrobial biofilms and inflammasome activation. A deeper understanding of the subgingival cytological (micro) landscape, the role of extracellular DNA (eDNA) during periodontitis, and contribution of the host immune eDNA to inflammasome persistence, may improve our understanding of the mechanisms underlaying severe forms of periodontitis.
UNASSIGNED: In this work, subgingival biolfilms developing on biologically neutral polyethylene terephthalate films placed in gingival cavities of patients with chronic periodontitis were investigated by confocal laser scanning microscopy (CLSM). This allowed examination of realistic cytological landscapes and visualization of extracellular polymeric substances (EPS) including amyloids, total proteins, carbohydrates and eDNA, as well as comparison with several single-strain in vitro model biofilms produced by oral pathogens such as Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus gordonii, S. sanguinis and S. mitis. Fluorescence in situ hybridization (FISH) analysis was also used to identify eDNA derived from eubacteria, streptococci and members of the Bacteroides-Porphyromonas-Prevotella (BPP) group associated with periodontitis.
UNASSIGNED: Analysis of subgingival biofilm EPS revealed low levels of amyloids and high levels of eDNA which appears to be the main matrix component. However, bacterial eDNA contributed less than a third of the total eDNA observed, suggesting that host-derived eDNA released in neutrophil extracellular traps may be of more importance in the development of biofilms causing periodontitis.
UNASSIGNED: eDNA derived from host immunocompetent cells activated at the onset of periodontitis may therefore be a major driver of bacterial persistence and pathogenesis.
UNASSIGNED: In this work, subgingival biolfilms developing on biologically neutral polyethylene terephthalate films placed in gingival cavities of patients with chronic periodontitis were investigated by confocal laser scanning microscopy (CLSM). This allowed examination of realistic cytological landscapes and visualization of extracellular polymeric substances (EPS) including amyloids, total proteins, carbohydrates and eDNA, as well as comparison with several single-strain in vitro model biofilms produced by oral pathogens such as Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus gordonii, S. sanguinis and S. mitis. Fluorescence in situ hybridization (FISH) analysis was also used to identify eDNA derived from eubacteria, streptococci and members of the Bacteroides-Porphyromonas-Prevotella (BPP) group associated with periodontitis.
UNASSIGNED: Analysis of subgingival biofilm EPS revealed low levels of amyloids and high levels of eDNA which appears to be the main matrix component. However, bacterial eDNA contributed less than a third of the total eDNA observed, suggesting that host-derived eDNA released in neutrophil extracellular traps may be of more importance in the development of biofilms causing periodontitis.
UNASSIGNED: eDNA derived from host immunocompetent cells activated at the onset of periodontitis may therefore be a major driver of bacterial persistence and pathogenesis.
摘要：
■已知牙周疾病与多微生物生物膜和炎性体活化有关。对龈下细胞学（微观）景观有了更深入的了解，细胞外DNA（eDNA）在牙周炎中的作用，以及宿主免疫eDNA对炎症小体持久性的贡献，可以提高我们对严重牙周炎的机制的理解。
■在这项工作中，通过共聚焦激光扫描显微镜（CLSM）研究了在慢性牙周炎患者牙龈腔中放置的生物中性聚对苯二甲酸乙二醇酯膜上形成的牙龈下生物膜。这允许检查真实的细胞学景观和细胞外聚合物质（EPS）的可视化，包括淀粉样蛋白，总蛋白质，碳水化合物和eDNA，以及与肺炎克雷伯菌等口腔病原体产生的几种单菌株体外模型生物膜的比较，铜绿假单胞菌,金黄色葡萄球菌,格氏链球菌,S、血统和米蒂斯。荧光原位杂交（FISH）分析也用于鉴定源自真细菌的eDNA，与牙周炎相关的链球菌和拟杆菌-卟啉-普雷沃氏菌（BPP）组的成员。
■龈下生物膜EPS的分析揭示了低水平的淀粉样蛋白和高水平的eDNA，这似乎是主要的基质成分。然而,细菌eDNA对所观察到的总eDNA的贡献不到三分之一，表明在中性粒细胞胞外陷阱中释放的宿主来源的eDNA可能在导致牙周炎的生物膜的发展中更为重要。
■来源于牙周炎发作时活化的宿主免疫活性细胞的eDNA因此可能是细菌持久性和发病机理的主要驱动因素。
■在这项工作中，通过共聚焦激光扫描显微镜（CLSM）研究了在慢性牙周炎患者牙龈腔中放置的生物中性聚对苯二甲酸乙二醇酯膜上形成的牙龈下生物膜。这允许检查真实的细胞学景观和细胞外聚合物质（EPS）的可视化，包括淀粉样蛋白，总蛋白质，碳水化合物和eDNA，以及与肺炎克雷伯菌等口腔病原体产生的几种单菌株体外模型生物膜的比较，铜绿假单胞菌,金黄色葡萄球菌,格氏链球菌,S、血统和米蒂斯。荧光原位杂交（FISH）分析也用于鉴定源自真细菌的eDNA，与牙周炎相关的链球菌和拟杆菌-卟啉-普雷沃氏菌（BPP）组的成员。
■龈下生物膜EPS的分析揭示了低水平的淀粉样蛋白和高水平的eDNA，这似乎是主要的基质成分。然而,细菌eDNA对所观察到的总eDNA的贡献不到三分之一，表明在中性粒细胞胞外陷阱中释放的宿主来源的eDNA可能在导致牙周炎的生物膜的发展中更为重要。
■来源于牙周炎发作时活化的宿主免疫活性细胞的eDNA因此可能是细菌持久性和发病机理的主要驱动因素。
