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通过桑格测序法分析种植体周围炎生物膜的微生物多样性

Microbiological diversity of peri-implantitis biofilm by Sanger sequencing.

作者信息

da Silva Ennyo S C, Feres Magda, Figueiredo Luciene C, Shibli Jamil A, Ramiro Fernanda S, Faveri Marcelo

机构信息

Department of Periodontology, Dental Research Division, Guarulhos University, Guarulhos, SP, Brazil.

出版信息

Clin Oral Implants Res. 2014 Oct;25(10):1192-9. doi: 10.1111/clr.12231. Epub 2013 Jul 12.

Abstract

BACKGROUND AND OBJECTIVE

To examine the microbial diversity associated with implants with or without peri-implantitis and to identify differences between the bacterial communities in these clinical conditions.

MATERIAL AND METHODS

Twenty subjects were assigned to a Control group consisting of subjects with healthy implants and a Test group consisting of subjects with peri-implantitis sites, as well as a healthy implant site (n = 10/group). In the Test group, subgingival biofilm samples were taken from the deepest sites of the diseased implant. In both groups, samples were collected from one site around a healthy implant. DNA was extracted and the 16S rRNA gene was amplified and sequenced, and the phylotypes were identified by comparison with known 16S rRNA sequences.

RESULTS

The phylogenetic identity of 1387 16S rRNA gene clones was determined. Healthy implants demonstrated higher proportions of Actinomyces, Atopobium, Gemella, Kingella and Rothia and lower levels of Campylobacter, Desulfobulbus, Dialister, Eubacterium, Filifactor, Mitsukella, Porphyromonas and Pseudoramibacter (Mann-Whitney U-test; P < 0.05). Fusobacterium nucleatum, Dialister invisus, Streptococcus sp. human oral taxon (HOT) 064, Filifactor alocis and Mitsuokella sp. HOT 131 presented a higher mean proportion, while Veillonella dispar, Actinomyces meyeri, Granulicatella adiacens showed lower mean proportions in the peri-implantitis sites when compared with healthy implants in both the Control and Test groups (Mann-Whitney U-test; P < 0.05).

CONCLUSION

Marked differences were observed in the composition of the subgingival biofilm between healthy and diseased implants. The biofilm associated with peri-implantitis harbored more pathogenic bacterial species from the orange complex and other "unusual" putative pathogens, such as F. alocis, D. invisus and Mitsuokella sp. HOT 131.

摘要

背景与目的

研究种植体周围有无种植体周围炎情况下的微生物多样性,并确定这些临床状况下细菌群落之间的差异。

材料与方法

20名受试者被分为两组,一组为对照组,由种植体健康的受试者组成;另一组为试验组,由有种植体周围炎部位以及一个健康种植体部位的受试者组成(每组n = 10)。试验组中,从患病种植体最深部位采集龈下生物膜样本。两组均从一个健康种植体周围的部位采集样本。提取DNA,扩增并测序16S rRNA基因,通过与已知16S rRNA序列比较鉴定系统发育型。

结果

确定了1387个16S rRNA基因克隆的系统发育特征。健康种植体中放线菌属、阿托波菌属、孪生球菌属、金氏菌属和罗氏菌属的比例较高,弯曲杆菌属、脱硫球菌属、纤毛菌属、真杆菌属、丝状杆菌属、米氏菌属、卟啉单胞菌属和假枝杆菌属的水平较低(曼-惠特尼U检验;P < 0.05)。与对照组和试验组的健康种植体相比,具核梭杆菌、隐匿纤毛菌、人口腔分类群(HOT)064链球菌、牙龈丝状杆菌和米氏菌属HOT 131在种植体周围炎部位的平均比例较高,而差异韦荣球菌、迈耶放线菌、毗邻颗粒链菌在种植体周围炎部位的平均比例较低(曼-惠特尼U检验;P < 0.05)。

结论

健康与患病种植体的龈下生物膜组成存在显著差异。与种植体周围炎相关的生物膜中含有更多来自橙色复合体的致病细菌种类以及其他“不常见”的假定病原体,如牙龈丝状杆菌、隐匿纤毛菌和米氏菌属HOT 131。

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