Sandros J, Papapanou P, Dahlén G
Department of Periodontology, Faculty of Odontology, University of Göteborg, Sweden.
J Periodontal Res. 1993 May;28(3):219-26. doi: 10.1111/j.1600-0765.1993.tb01072.x.
The aim of the present study was to analyze the adhesive and invasive potential of a number of P. gingivalis strains, in an in vitro system utilizing cultures of human oral epithelial cells (KB cell line, ATCC CCL 17). P. gingivalis strains W50 and FDC 381 (laboratory strains) and OMGS 1738, 1743 and 1439 (clinical isolates) as well as E. coli strain HB 101 (non-adhering, non-invasive control) were used. Adherence was assessed by means of scintillation counting and light microscopy, after incubation of radiolabelled bacteria with epithelial cells. In the invasion assay, monolayers were infected with the P. gingivalis and E. coli strains and further incubated with an antibiotic mixture (metronidazole 0.1 mg/ml and gentamicin 0.5 mg/ml). Invasion was evaluated by (i) assessing presence of bacteria surviving the antibiotic treatment, and (ii) electron microscopy. All P. gingivalis strains adhered to and entered into the oral epithelial cells. After 3 hours of incubation, bacteria were frequently identified intracellularly by means of electron microscopy. The cellular membranes, encapsulating the microorganisms in early stages of the invasive process, appeared later to disintegrate. The presence of coated pits on the epithelial cell surfaces suggested that internalization of P. gingivalis was associated with receptor-mediated endocytosis (RME). Formation of outer membrane vesicles (blebs) by intracellular bacteria indicated that internalized P. gingivalis was able to retain its viability. E. coli strain HB 101 neither adhered to nor invaded epithelial cells.
本研究的目的是在利用人口腔上皮细胞(KB细胞系,ATCC CCL 17)培养物的体外系统中,分析若干牙龈卟啉单胞菌菌株的黏附及侵袭潜能。使用了牙龈卟啉单胞菌菌株W50和FDC 381(实验室菌株)、OMGS 1738、1743和1439(临床分离株)以及大肠杆菌菌株HB 101(非黏附、非侵袭对照)。在用放射性标记的细菌与上皮细胞孵育后,通过闪烁计数和光学显微镜评估黏附情况。在侵袭试验中,用牙龈卟啉单胞菌和大肠杆菌菌株感染单层细胞,并进一步与抗生素混合物(甲硝唑0.1 mg/ml和庆大霉素0.5 mg/ml)一起孵育。通过(i)评估在抗生素处理后存活的细菌的存在情况以及(ii)电子显微镜来评估侵袭情况。所有牙龈卟啉单胞菌菌株均黏附并进入口腔上皮细胞。孵育3小时后,通过电子显微镜经常在细胞内鉴定到细菌。在侵袭过程早期包裹微生物的细胞膜后来似乎解体。上皮细胞表面有被膜小窝表明牙龈卟啉单胞菌的内化与受体介导的内吞作用(RME)有关。细胞内细菌形成外膜囊泡(泡)表明内化的牙龈卟啉单胞菌能够保持其活力。大肠杆菌菌株HB 101既不黏附也不侵袭上皮细胞。