Division of Periodontics, Department of Stomatology, School of Dentistry, University of São Paulo, São Paulo, Brazil.
Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
J Periodontal Res. 2019 Apr;54(2):115-127. doi: 10.1111/jre.12608. Epub 2018 Oct 4.
Although previous studies revealed the potential use of probiotics in the control of periodontitis, little is known about their interactions with gingival epithelial cells (GECs). Since GECs comprise the first defense in the subgingival microenvironment, the aim of this study was to evaluate the effect of probiotic lactobacilli and bifidobacteria strains on OBA-9 cells challenged with Porphyromonas gingivalis.
Immortalized human GECs (OBA-9) were challenged with live P. gingivalis (strains W83 and ATCC33277) and co-infected with one of 12 tested probiotic strains at a multiplicity of infection (MOI) of 1:1000 for 2 hours. Bacterial adhesion and invasion were determined by antibiotic exclusion analysis and CFU counting. OBA-9 viability was assessed by MTT assay, and levels of inflammatory mediators (TNF-α, IL-1β, and CXCL8) in the supernatants were determined by ELISA. The expression of genes encoding Toll-like receptors (TLR2, TLR4) was evaluated by RT-qPCR.
Both strains of P. gingivalis were able to adhere and invade OBA-9 cells, with significant loss in cell viability, increase in the levels of TNF-α and IL-1β, and upregulation of TLR4. However, co-infection with probiotics attenuated these effects in P. gingivalis challenged GECs. Most probiotics maintained OBA-9 viability and reduced pathogens adhesion and invasion. Furthermore, probiotics were able to adhere to GECs, which was enhanced for most strains in the presence of P. gingivalis. The synthesis of IL-1β and TNF-α by P. gingivalis in challenged GECs was reduced in co-culture with most of the tested probiotics, whereas the secretion of CXCL8 increased, and TLR4 was downregulated.
Probiotics can alter the interaction of GECs with P. gingivalis by modulating the pathogen's ability to adhere and invade these cells, as well as by regulating the innate immune response. Such properties are strain-specific and may indicate the most efficient probiotics to control periodontitis.
虽然先前的研究揭示了益生菌在控制牙周炎方面的潜在用途,但人们对它们与牙龈上皮细胞(GECs)的相互作用知之甚少。由于 GECs 构成了龈下微环境中的第一道防线,因此本研究旨在评估益生菌乳杆菌和双歧杆菌菌株对受牙龈卟啉单胞菌挑战的 OBA-9 细胞的影响。
将永生化人 GEC(OBA-9)用活的牙龈卟啉单胞菌(菌株 W83 和 ATCC33277)进行挑战,并以感染复数(MOI)为 1:1000 将其与 12 种测试的益生菌菌株之一共感染 2 小时。通过抗生素排除分析和 CFU 计数来确定细菌黏附和侵袭。通过 MTT 测定评估 OBA-9 细胞活力,通过 ELISA 测定上清液中炎症介质(TNF-α、IL-1β 和 CXCL8)的水平。通过 RT-qPCR 评估编码 Toll 样受体(TLR2、TLR4)的基因的表达。
两种牙龈卟啉单胞菌菌株均能黏附和侵袭 OBA-9 细胞,导致细胞活力显著下降,TNF-α 和 IL-1β 水平升高,TLR4 上调。然而,与益生菌共感染可减轻受牙龈卟啉单胞菌挑战的 GEC 中的这些作用。大多数益生菌维持 OBA-9 细胞活力并减少病原体黏附和侵袭。此外,益生菌能够黏附在 GEC 上,而在存在牙龈卟啉单胞菌的情况下,大多数菌株的黏附能力增强。在与大多数测试的益生菌共培养时,受挑战的 GEC 中牙龈卟啉单胞菌合成的 IL-1β 和 TNF-α 减少,而 CXCL8 的分泌增加,TLR4 下调。
益生菌可以通过调节病原体黏附和侵袭这些细胞的能力以及调节先天免疫反应来改变 GEC 与牙龈卟啉单胞菌的相互作用。这些特性是菌株特异性的,可能表明最有效的益生菌来控制牙周炎。