Extracrevicular Invasion by Periodontal Pathogens

牙周病原体的沟外侵袭

• 批准号: 6917237
• 负责人: Joel D. Rudney
• 金额: $ 26.07万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6917237
• 关键词: Actinobacillus actinomycetemcomitans; Bacteroides gingivalis; atherosclerotic plaque; bacteria infection mechanism; chlorhexidine; clinical research; confocal scanning microscopy; cytokine; denaturing gradient gel electrophoresis; disease /disorder proneness /risk; fluorescent in situ hybridization; gene expression; green fluorescent proteins; human subject; messenger RNA; oral mucosa; periodontitis; polymerase chain reaction; ribosomal RNA; saliva; stroke; tissue /cell culture

Gingival infection with periodontal pathogens often persists after treatment, and mucosa are thought to be a reservoir for recolonization. Invaded mucosal cells may provide a protected environment for these fastidious anaerobes. Preliminary studies used fluorescent in situ hybridization (FISH) with universal and specific rRNA probes and confocal microscopy (LSCM) to detect Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and unidentified bacteria inside buccal cells from 23 of 24 subjects. This suggests that intracellular mucosal bacteria exist in a multi-species community which may maintain itself by modulating or overcoming host cell defenses, with exfoliated cells providing a protected route for bacterial transmission. Those postulates will be tested by these specific aims: 1.) Determine the composition of the mucosal intracellular community by probing for additional periodontal pathogens and other mucosal species. 16S rRNA temperature gradient electrophoresis will be used to design probes for uncultured species. Multicolor FISH will determine whether more than one species can occupy a cell. 2.) Determine whether intracellular bacteria modulate or overcome cell defenses in vivo by using multicolor FISH/LCSM to compare invaded and uninvaded cells for expression of cytokine and antimicrobial peptide mRNA. 3.) Determine whether exfoliated invaded cells may be a vector for transmission by using a tissue culture model. Washed mucosal cells will be suspended in autologous or heterologous clarified saliva with or without antibiotics, and then incubated with green fluorescent protein (GFP)-labeled KB cells. Transmission will be evaluated by multicolor FISH/LSCM. 4.) Determine whether the intracellular mucosal community can establish and maintain itself in the absence of the gingival crevice by using FISH/LSCM to look for such communities in predentate infants, and to verify whether they occur in edentulous adults. 5.) Determine whether mucosal invasion protects pathogens from elimination in periodontal patients who require aggressive treatment, by using FISH/LSCM and multiplex PCR to compare mucosal and gingival colonization before and after scaling and root planing, topical chlorhexidine, and systemic antibiotics.

牙周病原体引起的牙龈感染通常在治疗后仍然存在，而粘膜被认为是重新定植的储存库。侵入的粘膜细胞可能为这些挑剔的厌氧菌提供了一个受保护的环境。初步研究使用荧光原位杂交（FISH）和通用和特异性rRNA探针和共聚焦显微镜（LSCM）检测了24名受试者中的23名口腔细胞内的放线菌、牙龈卟啉单胞菌和未识别的细菌。这表明细胞内粘膜细菌存在于一个多物种群落中，它们可能通过调节或克服宿主细胞防御来维持自身，脱落的细胞为细菌传播提供了一条受保护的途径。这些假设将受到以下具体目标的检验：通过探测额外的牙周病原体和其他粘膜物种来确定粘膜细胞内群落的组成。16S rRNA温度梯度电泳将用于设计未培养物种的探针。多色FISH将决定是否有多个物种可以占据一个细胞。2)。通过使用多色FISH/LCSM比较入侵和未入侵细胞中细胞因子和抗菌肽mRNA的表达，确定细胞内细菌是否调节或克服体内细胞防御。3)。通过组织培养模型确定脱落的入侵细胞是否可以作为传播载体。将清洗后的粘膜细胞悬浮在有或不含抗生素的自体或异源澄清唾液中，然后与绿色荧光蛋白（GFP）标记的KB细胞孵育。透射率将通过多色FISH/LSCM进行评估。4.） 通过FISH/LSCM在有牙的婴儿中寻找细胞内粘膜群落，并验证它们是否发生在无牙的成人中，确定细胞内粘膜群落是否可以在没有龈缝的情况下建立和维持自身。5)。在需要积极治疗的牙周患者中，通过FISH/LSCM和多重PCR来比较刮治和根治、局部氯己定和全身抗生素前后的粘膜和牙龈定植情况，以确定粘膜侵入是否保护病原体不被清除。

项目成果

Oral bacteria in plaque and invading buccal cells of young orthodontic patients.

• DOI: 10.1016/j.ajodo.2006.05.028
• 发表时间: 2006-12
• 期刊: American journal of orthodontics and dentofacial orthopedics : official publication of the American Association of Orthodontists, its constituent societies, and the American Board of Orthodontics
• 作者: Nancy M Leung;Ruoqiong Chen;J. Rudney

Persistence of extracrevicular bacterial reservoirs after treatment of aggressive periodontitis.

• DOI: 10.1902/jop.2008.080254
• 发表时间: 2008-12
• 期刊: JOURNAL OF PERIODONTOLOGY
• 影响因子: 4.3
• 作者: Johnson, Jason D.;Chen, Ruoqiong;Lenton, Patricia A.;Zhang, Guizhen;Hinrichs, James E.;Rudney, Joel D.
• 通讯作者: Rudney, Joel D.

Streptococcus cristatus attenuates Fusobacterium nucleatum-induced interleukin-8 expression in oral epithelial cells.

冠状链球菌减弱具核梭杆菌诱导的口腔上皮细胞中白细胞介素 8 的表达。

• DOI: 10.1111/j.1600-0765.2007.01057.x
• 发表时间: 2008
• 期刊: Journal of periodontal research
• 影响因子: 3.5
• 作者: Zhang,G;Chen,R;Rudney,JD
• 通讯作者: Rudney,JD