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

本研究旨在观察人类牙周袋内某些菌斑细菌的定位和分布，包括牙周病相关细菌。在提供知情同意后，对患有晚期成人牙周炎的牙齿及其周围牙周组织进行仔细检查，尽量不改变牙周袋的结构。将其加工成连续切片，用改良革兰氏染色法和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体染色。光镜下观察人牙周袋内细菌种类的分布和定位。少量牙龈卟啉单胞菌散在牙周袋内。艾肯菌锈菌主要分布在中深袋区根表面附近，形成小块状。多核梭杆菌在中间和深袋区也有小团块，但其定位在未附着的斑块区。直弯杆菌一般在牙周袋的中部和深部形成大团块，位于牙周袋的根表面和上皮表面。未附着斑块区主要位于囊袋中部，可见密螺旋体。在此之前，我们发现e.c rodens 1073具有细胞相关的凝集素样物质（EcLS），它在牙周病相关细菌粘附在各种细胞表面的过程中起作用。利用单克隆抗体对EcLS进行免疫电镜研究，发现EcLS定位于外膜外空间，而不是在细菌具有粘附素的细胞表面附属物如菌毛中。当与BALB/c小鼠的B淋巴细胞培养时，发现该EcLS具有强大的有丝分裂活性。EcLS不仅能刺激小鼠B淋巴细胞增殖，还能向抗体分泌细胞分化。这些发现表明，EcLS可以刺激宿主细胞，而不仅仅是介导宿主细胞与e.o rodens之间的粘附。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