Analysis of bacterial biofilms as a virulence factor for peri-implantitis

我们调查了形态学上的初始生物膜形成的人类种植沟愈合基台。在获得7名患者的知情同意后，在插入后的第10、30、90和150天小心地移除20个愈合基台，并进行扫描电子显微镜检查。在第10天，单层生物膜已部分形成在基牙表面上。观察到所谓的早期殖民者：球菌和丝状菌。30天后，生物膜形成，在人类种植体沟中主要观察到长杆和短杆。取4例患者的8枚种植体，取半薄切片，用4种抗牙周病菌抗体进行BrownBrenn改良革兰氏染色和链霉亲和素生物素法染色。在种植体表面和基台螺钉内均检测到革兰氏阳性菌，在人种植体龈沟中未检测到种属特异性抗体标记的牙龈卟啉单胞菌和直弯曲杆菌。（3）检测了生物膜形成所必需的基因（多磷酸激酶（ppk）的同源物）。生物膜中ATP含量的增加率很低，为正常细胞的1/3.0 × 10 ~ 5。葡萄糖酸氯己定可有效降低牙龈卟啉单胞菌生物膜的活力，而盐酸米诺环素和甲硝唑则无可变效应。所克隆的牙龈卟啉单胞菌ppk基因与大肠杆菌ppk基因的同源性为35.8%，其中牙龈卟啉单胞菌ppk基因高度保守

We investigated morphologically initial biofilm formation in the human implant sulcus using healing abutments. After obtaining informed consent from 7 patients, twenty healing abutments were removed carefully on day 10, 30, 90 and 150 after insertion and treated for a scanning electron microscopy. On day 10, mono-layer biofilm have partially formed on the abutment surfaces. So called early colonizers: cocci and filaments, were observed. After 30 days, biofilms developed, and long and short rods were predominantly observed in human implant sulcusNext, human periimplantitis associated bacteria have been examined immunohistochemically. Eight implants obtained from 4 voluntary patients were extracted and processed into semithin sections They were stained BrownBrenn modified Gram stain and streptavidinbiotin method using 4 kinds of antibodies with periodontopathic bactria. Gram positive bacteria were shown not only on the implant surface but also at the inner site of abutment screw, Porphyromonas gingivalis and Campylobacter rectus labeled with speciesspecific antibody were not found in the human implant sulcusBy use of the in vitro model with P. gingivalis biofilm, 1) the biological (ATP) activities, 2) the susceptibility to antimicrobial agents, and 3) the essential gene for development of biofilm (the homologue for polyphosphate kinase (ppk)) were examined. The rate of increase of ATP contents in the biofilm was quite low, and 1/3.0x10^5 compared with that of planktonic cells. Chlorhexidine gluconate was effective at reducing the viability of P. gingivalis biofilm, while minocycline hydrochloride and metronidazole did not have a variable effect. Identity of P. gingivalis ppk gene that was carried out cloning to Esherihia coli ppk gene was 35.8%, including \hat ppk from P. gingivalis was highly conserved