Department of Periodontology and Operative Dentistry, Münster University Hospital, Münster, Germany.
Department of Periodontology, Johann Wolfgang Goethe-University Frankfurt, Frankfurt, Germany.
Front Cell Infect Microbiol. 2020 May 5;10:167. doi: 10.3389/fcimb.2020.00167. eCollection 2020.
The aim of this follow-up study was, to compare the effects of mechanical periodontal therapy with or without adjunctive amoxicillin and metronidazole on the subgingival microbiome of smokers with periodontitis using 16S rDNA amplicon next generation sequencing. Fifty-four periodontitis patients that smoke received either non-surgical periodontal therapy with adjunctive amoxicillin and metronidazole (n = 27) or with placebos (n = 27). Subgingival plaque samples were taken before and two months after therapy. Bacterial genomic DNA was isolated and the V4 hypervariable region of the bacterial 16S rRNA genes was amplified. Up to 96 libraries were normalized and pooled for Illumina MiSeq paired-end sequencing with almost fully overlapping 250 base pairs reads. Exact ribosomal sequence variants (RSVs) were inferred with DADA2. Microbial diversity and changes on the genus and RSV level were analyzed with non-parametric tests and a negative binomial regression model, respectively. Before therapy, the demographic, clinical, and microbial parameters were not significantly different between the placebo and antibiotic groups. Two months after the therapy, clinical parameters improved and there was a significantly increased dissimilarity of microbiomes between the two groups. In the antibiotic group, there was a significant reduction of genera classified as , and , and 22 other genera also decreased significantly, while , and five other genera significantly increased. In the placebo group, however, there was not a significant decrease in periodontal pathogens after therapy and only five other genera decreased, while and nine other genera increased. We conclude that in periodontitis patients who smoke, microbial shifts occurred two months after periodontal therapy with either antibiotics or placebo, but genera including periodontal pathogens decreased significantly only with adjunctive antibiotics.
本随访研究旨在比较机械牙周治疗联合或不联合阿莫西林和甲硝唑辅助治疗对吸烟牙周炎患者龈下微生物组的影响,方法是使用 16S rDNA 扩增子下一代测序。54 名吸烟的牙周炎患者分别接受非手术牙周治疗联合阿莫西林和甲硝唑辅助治疗(n = 27)或安慰剂治疗(n = 27)。在治疗前和治疗后两个月采集龈下菌斑样本。提取细菌基因组 DNA,扩增细菌 16S rRNA 基因 V4 高变区。多达 96 个文库进行归一化,并混合进行 Illumina MiSeq 配对末端测序,得到几乎完全重叠的 250 个碱基对读数。使用 DADA2 推断精确核糖体序列变体(RSV)。使用非参数检验和负二项式回归模型分别分析微生物多样性和属及 RSV 水平的变化。治疗前,安慰剂和抗生素组之间的人口统计学、临床和微生物参数无显著差异。治疗 2 个月后,临床参数改善,两组之间的微生物组差异显著增加。在抗生素组中,属于 、 和 的属显著减少,22 个其他属也显著减少,而 、 和 5 个其他属显著增加。然而,在安慰剂组中,治疗后牙周病致病菌并没有显著减少,只有 5 个其他属减少,而 、 和 9 个其他属增加。我们的结论是,在吸烟的牙周炎患者中,牙周治疗后两个月会发生微生物转移,无论是联合抗生素还是安慰剂治疗,但只有联合抗生素治疗才能显著减少包括牙周病致病菌在内的属。