In Vitro Effect of Porphyromonas gingivalis Methionine Gamma Lyase on Biofilm Composition and Oral Inflammatory Response.
In Vitro Effect of Porphyromonas gingivalis Methionine Gamma Lyase on Biofilm Composition and Oral Inflammatory Response.
复制标题
DOI：
10.1371/journal.pone.0169157
复制
发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Allaker RP
中科院分区：
文献类型：
--
作者：
Stephen AS;Millhouse E;Sherry L;Aduse-Opoku J;Culshaw S;Ramage G;Bradshaw DJ;Burnett GR;Allaker RP
甲硫醇（甲硫醇）是口腔恶臭和牙周组织破坏的重要因素。牙龈卟啉单胞菌、中间普雷沃菌和具核梭杆菌是通过甲硫氨酸γ裂解酶（mgl）活性产生甲硫醇的关键口腔微生物菌种。本研究的目的是使用具有口腔角质形成细胞的10种生物膜共培养模型比较牙龈卟啉单胞菌的mgl敲除菌株与其野生型及其对生物膜组成和炎性细胞因子产生的影响。使用来自野生型W50的插入诱变构建牙龈卟啉单胞菌mgl敲除菌株，气相色谱顶空分析证实缺乏甲硫醇产生。10-由缓症链球菌、口腔链球菌、中间链球菌、具核梭杆菌多形亚种、具核梭杆菌文森特亚种、韦荣氏球菌、内氏放线菌、中间普雷沃氏菌和伴放线菌聚集杆菌与野生型或突变体牙龈卟啉单胞菌组成的物种生物膜在Thermanox盖玻片上生长，并用于刺激口腔角质形成细胞（OKF 6-TERT 2），在厌氧条件下培养4和24小时。用SYBR绿色通过定量PCR分析生物膜的微生物生态学变化。使用细胞因子的多重珠免疫测定法分析角质细胞培养上清液。在突变体和野生型生物膜之间观察到显著的群体差异;舞毒菌比例增加（p<0.001），而A.内氏链球菌（P<0.01）和链球菌（P <0.01）。（p<0.05）。与野生型相比，当用突变体生物膜刺激时，角质形成细胞产生较少的IL-8、IL-6和IL-1α。牙龈卟啉单胞菌中缺乏mgl已显示影响体外微生物生态，产生显著不同的生物膜组成，观察到来自角质形成细胞的更多促炎细胞因子应答。甲硫醇在生物膜形成和细胞因子反应中可能发挥作用，并对口腔恶臭和牙周炎产生影响。
Methanethiol (methyl mercaptan) is an important contributor to oral malodour and periodontal tissue destruction. Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum are key oral microbial species that produce methanethiol via methionine gamma lyase (mgl) activity. The aim of this study was to compare an mgl knockout strain of P. gingivalis with its wild type using a 10-species biofilm co-culture model with oral keratinocytes and its effect on biofilm composition and inflammatory cytokine production. A P. gingivalis mgl knockout strain was constructed using insertion mutagenesis from wild type W50 with gas chromatographic head space analysis confirming lack of methanethiol production. 10-species biofilms consisting of Streptococcus mitis, Streptococcus oralis, Streptococcus intermedius, Fusobacterium nucleatum ssp polymorphum, Fusobacterium nucleatum ssp vincentii, Veillonella dispar, Actinomyces naeslundii, Prevotella intermedia and Aggregatibacter actinomycetemcomitans with either the wild type or mutant P. gingivalis were grown on Thermanox cover slips and used to stimulate oral keratinocytes (OKF6-TERT2), under anaerobic conditions for 4 and 24 hours. Biofilms were analysed by quantitative PCR with SYBR Green for changes in microbial ecology. Keratinocyte culture supernatants were analysed using a multiplex bead immunoassay for cytokines. Significant population differences were observed between mutant and wild type biofilms; V. dispar proportions increased (p<0.001), whilst A. naeslundii (p<0.01) and Streptococcus spp. (p<0.05) decreased in mutant biofilms. Keratinocytes produced less IL-8, IL-6 and IL-1α when stimulated with the mutant biofilms compared to wild type. Lack of mgl in P. gingivalis has been shown to affect microbial ecology in vitro, giving rise to a markedly different biofilm composition, with a more pro-inflammatory cytokine response from the keratinocytes observed. A possible role for methanethiol in biofilm formation and cytokine response with subsequent effects on oral malodor and periodontitis is suggested.
登录
查看更多内容
影响因子：
64.8
作者：
Bettelli, E;Carrier, YJ;Kuchroo, VK
通讯作者：
Kuchroo, VK
Keystone-Pathogen假设。
DOI：
10.1038/nrmicro2873
发表时间：
2012-10
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
通讯作者：
--
影响因子：
3.8
作者：
Hamedi, M.;Belibasakis, G. N.;Bostanci, N.
通讯作者：
Bostanci, N.
影响因子：
4.8
作者：
Calkins, CC;Platt, K;Travis, J
通讯作者：
Travis, J
影响因子：
2.6
作者：
GHARBIA, SE;SHAH, HN;WELCH, SG
通讯作者：
WELCH, SG