Localization of periodontal disease-associated bacteria in human periodontal pockets and its relationship to tissue destruction

本研究旨在观察包括牙周病相关细菌在内的某些菌斑细菌在人牙周袋中的定位和分布。在提供知情同意后，仔细获取患有晚期成人牙周炎的牙齿及其周围的牙周组织，尽量不改变牙周袋的结构。连续切片，用Brown-Brenn改良革兰氏染色和LASB法对包括牙周病相关细菌在内的15种菌斑细菌进行特异性兔抗菌染色。在光镜下观察牙周袋内细菌的分布和定位。牙龈卟啉单胞菌的小聚集物散布在牙周袋中。腐蚀型艾氏球藻主要分布在中袋和深袋的根面附近，形成小团块。梭杆菌…更多的核仁在中袋和深袋也有小团块，但其定位在未附着的斑块区。而直弯杆菌一般在牙周袋的中深部形成大团块，紧密分布在牙周袋的根面和上皮面。齿密螺旋体主要分布在菌斑的中部，先前我们发现腐蚀性螺旋体1073有一种细胞相关凝集素样底物(ECLS)，它在牙周病相关细菌与多种细胞表面的黏附中起作用。用抗ECLS的单抗进行免疫电子显微镜研究表明，ECLS定位于外膜的外层空间，而不是定位于许多细菌具有粘附素的菌毛等细胞表面附属物。这种ECLS与BALB/c小鼠的B淋巴细胞培养后具有较强的促分裂活性。ECLS不仅能刺激小鼠B淋巴细胞增殖，而且能分化为抗体分泌细胞。这些发现表明，ECLS可以刺激宿主细胞，而不仅仅是调节宿主细胞和E.腐蚀之间的黏附。较少

This study was designed to observe the localization and distribution of certain plaque bacteria including periodontal disease-associated bacteria in human periodontal pocket. After providing informed consent, teeth and their surrounding periodontal tissue with advanced adult periodontitis were obtained carefully attempting not to change the structure of the periodontal pockets. They were processed into serial sections, and those were stained with Brown-Brenn modified Gram stain and LASB method with specific rabbit antiboeies against fifteen plaque bacterial species including periodontal disease-associated bacteria. The localization and distribution of the bacterial species in human periodontal pockets were observed under a light microscope. A small aggregates of Porphyromonas gingivalis were distributed spattering throughout the periodontal pocket. Eikenella corrodens, shich formed small clumps, was located mainly close to root surface in the middle and deep pocket zones. Fusobacterium … More nucleatum also consisted small clumps in the middle and deep pocket zones, but its localization was in the unattached plaque area. On the other hand, Camphylobacter rectus generaly formed large clumps in the middle and deep pocket zones, and located closely on both root surface and epithelial surface in periodontal pockets. Treponema denticola was observed over the unattached plaque area located mainly in the middle zone of pockets.previously, we found that E.corrodens 1073 had a cell-associated lectin-like substans (EcLS) and it functioned in the adherence of this periodontal disease-associated bacteria to various cell surface. The immunoelectron microscopic study using monoclonal antibody against EcLS revealed that EcLS was localized in the outer space of outer membrane, not in cell surface appendages such as fimbriae where many bacteria possessed adhesin. This EcLS was found to have potent mitogenic activity when cultured with B lymphocytes from BALB/c mice. EcLS stimulated murine B lymphocytes not only to proliferate, but also to differentiate into antibody-secreting cells. These findings suggest that EcLS can stimulate host cells, and does not just mediate adherence between the host cell and E.corrodens. Less