Section of Preventive Dentistry, Department of Conservative and Preventive Dentistry, Academic Centre for Dentistry Amsterdam, ACTA, University of Amsterdam and Vrije University Amsterdam, Amsterdam, The Netherlands.
Arch Immunol Ther Exp (Warsz). 2012 Dec;60(6):469-76. doi: 10.1007/s00005-012-0196-8. Epub 2012 Sep 5.
Porphyromonas gingivalis is a Gram-negative, anaerobic bacterium involved in periodontitis and peri-implantitis that can invade and survive inside host cells in vitro. P. gingivalis can invade human gingival fibroblasts (GF), but no data are available about the role of P. gingivalis' capsule in GF invasion. In the current study, we aimed to determine the ability of three strains of P. gingivalis (encapsulated wild type W83, non-encapsulated HG91 and the non-encapsulated insertional isogenic knockout mutant of W83, ΔEpsC) to invade GF and the ability of internalized P. gingivalis to survive in vitro antibiotic treatment. The ability of P. gingivalis strains to invade GF was tested using an antibiotic protection assay at multiplicity of infection (MOI) 100 and 1000. The survival of internalized P. gingivalis cells was further analyzed by subsequent in vitro treatment with either metronidazole or amoxicillin alone or a combination of metronidazole and amoxicillin and anaerobic culture viability counts. All strains of P. gingivalis used in this study were able to invade GFs. The non-encapsulated mutant of W83 (ΔEpsC mutant) was significantly more invasive than the wild type W83 at MOI 100 (p value 0.025) and MOI 1000 (p value 0.038). Furthermore, internalized P. gingivalis was able to resist in vitro antibiotic treatment. As demonstrated by the differences in invasion efficiencies of P. gingivalis strain W83 and its isogenic mutant ΔEpsC, the capsule of P. gingivalis makes it less efficient in invading gingival fibroblasts. Moreover, internalized P. gingivalis can survive antibiotic treatment in vitro.
牙龈卟啉单胞菌是一种革兰氏阴性、厌氧细菌,与牙周炎和种植体周围炎有关,能够在体外侵入并在宿主细胞内存活。牙龈卟啉单胞菌可以侵入人牙龈成纤维细胞(GF),但目前尚无关于牙龈卟啉单胞菌荚膜在 GF 入侵中的作用的数据。在本研究中,我们旨在确定三种牙龈卟啉单胞菌菌株(包膜野生型 W83、无包膜 HG91 和 W83 的非包膜插入同源缺失突变体ΔEpsC)侵入 GF 的能力,以及内化的牙龈卟啉单胞菌在体外抗生素治疗中的存活能力。使用抗生素保护测定法在感染复数(MOI)为 100 和 1000 时测试了牙龈卟啉单胞菌菌株侵入 GF 的能力。通过随后用单独的甲硝唑或阿莫西林或甲硝唑和阿莫西林联合治疗以及厌氧培养存活计数,进一步分析内化的牙龈卟啉单胞菌细胞的存活情况。本研究中使用的所有牙龈卟啉单胞菌菌株都能够侵入 GFs。W83 的无包膜突变体(ΔEpsC 突变体)在 MOI 100(p 值 0.025)和 MOI 1000(p 值 0.038)时比野生型 W83 更具侵袭性。此外,内化的牙龈卟啉单胞菌能够抵抗体外抗生素治疗。正如牙龈卟啉单胞菌菌株 W83 及其同源缺失突变体ΔEpsC 的入侵效率差异所表明的那样,牙龈卟啉单胞菌的荚膜使其侵入牙龈成纤维细胞的效率降低。此外,内化的牙龈卟啉单胞菌可以在体外抗生素治疗中存活。