Influence of mechanical debridement on the subgingival microbiome in chronic periodontitis

doi:10.12016/j.issn.2096-1456.2021.06.002

Abstract

Abstract:

Objective To investigate the influence of mechanical debridement on the subgingival microbiome in chronic periodontitis by 16S rRNA high-throughput sequencing. Methods Patients with generalized moderate to severe chronic periodontitis were recruited into this study and received oral hygiene instruction and supragingival scaling. One week later, they received ultrasonic and manual subgingival scaling and root planning. Clinical parameters were recorded and subgingival plaques were sampled at baseline and 3 months and 6 months after treatment. The comprehensive profiles of the subgingival microbiome were analyzed by sequencing the V3-4 region of 16S rRNA with the Illumina MiSeq platform. Results Alpha diversity analysis showed that the richness and diversity of the subgingival community were consistent before and after treatment, but a significant difference in community structure was detected only between baseline and month 3 by principal coordinates analysis (PCoA). After 3 months, the clinical parameter as probing depth (PD) decreased significantly and the relative abundances of the genera related to periodontitis such as Porphyromonas, Treponema, Tannerella, and Filifactor decreased significantly. Meanwhile, the relative abundances of the genera associated with periodontal health increased, such as Capnocytophaga, Kingella. Six months later, however, less genera related to periodontitis decreased significantly from the baseline level, such as Filifactor. PD decreased significantly compared with baseline, but increased significantly compared with 3 months after treatment. Conclusion Mechanical debridement alone could relieve periodontal inflammation and balance microbial dysbiosis and the greater efficacy occurred 3 months after treatment.

Key words: chronic periodontitis, subgingival microbiome, dysbacteriosis, mechanical debridement, subgingival scaling and root planing, high-throughput sequencing, 16S rRNA, longitudinal survey

摘要：

目的采用16S rRNA高通量测序技术,探讨单纯机械治疗对慢性牙周炎龈下菌群微生态的影响。方法纳入广泛型中重度慢性牙周炎患者,接受口腔卫生宣教,并同时进行龈上洁治。一周后进行超声结合手工龈下刮治及根面平整,在基线、治疗后3个月和治疗后6个月,记录临床指标并采集龈下菌斑。利用Illumina MiSeq平台进行16S rRNA V3-4区测序,比较治疗前后临床指标及微生物群落的特征变化。结果 Alpha多样性分析显示各时间点样本中微生物多样性和丰富度无明显变化,PCoA显示仅治疗后3个月与基线样本微生物群落结构存在显著性差异。治疗后3个月,临床指标探诊深度显著减小,卟啉单胞菌属、密螺旋体属、坦纳氏菌属、产线菌属等致病相关微生物的相对丰度显著降低,二氧化碳噬纤维菌属、金氏菌属等牙周健康相关微生物的相对丰度显著升高;治疗后6个月,虽产线菌属等致病相关微生物的相对丰度仍较基线显著降低,但种类数量较3个月时减少;探诊深度较基线明显降低,但较治疗后3个月显著增加。结论单纯机械治疗有助于缓解牙周炎症并改善龈下菌群的失调状态,在治疗后3个月内治疗效果较为显著。

关键词: 慢性牙周炎, 龈下菌群微生态, 菌群失调, 机械治疗, 龈下刮治及根面平整, 高通量测序, 16S rRNA, 纵向观察

CLC Number:

WANG Yanfeng,ZENG Jiajun,YUAN Qiao,LUAN Qingxian. Influence of mechanical debridement on the subgingival microbiome in chronic periodontitis[J]. Journal of Prevention and Treatment for Stomatological Diseases, 2021, 29(6): 368-376.

王延峰,曾佳骏,袁乔,栾庆先. 单纯机械治疗对慢性牙周炎患者龈下菌群微生态的影响[J]. 口腔疾病防治, 2021, 29(6): 368-376.

Figures/Tables 7

Table 1 Changes in the clinical parameters of patients with chronic periodontitis before and after treatment

Parameters	Baseline	3 months	6 months
PD（all sites,n=233,mm）	5.27 ± 1.37	3.49 ± 1.27^1）	3.67 ± 1.33^1）2）
PD（sampling sites,n=12,mm）	6.33 ± 1.30	3.58 ± 1.08^1）	4.00 ± 1.26^1）
BI	2.67 ± 0.81	1.90 ± 0.80^1）	1.82 ± 0.68^1）
PLI	0.93 ± 0.70	0.65 ± 0.65^1）	0.61 ± 0.70^1）

Table 2 Microbial richness and diversity of subgingival samples before and after mechanical debridement

Time	Chao 1	Observed species	Shannon index
Baseline	306.23 ± 35.09	248.98 ± 33.73	4.71 ± 0.44
3 months	325.83 ± 31.44	265.07 ± 28.06	4.81 ± 0.59
6 months	326.27 ± 49.14	262.48 ± 47.60	4.68 ± 0.32

Figure 1 Principal coordinates analysis of subgingival samples before and after treatment

Figure 2 Mean relative abundance of dominant phyla before and after treatment

Figure 3 Mean relative abundance at the genus level before and after treatment a: the 25 most abundant genera; b: genera with a significant difference before and after treatment; *: 3 months versus baseline, P < 0.05; #: 6 months versus baseline, P < 0.05

Figure 4 Mean relative abundance at the species level before and after treatment a: the 25 most abundant genera; b&c: genera with significant differences before and after treatment; *: 3 months versus baseline, P < 0.05; #: 6 months versus baseline, P < 0.05; §: 6 months versus 3 months, P < 0.05

Figure 5 Microbial analysis based on LEfSe method Microbial cladogram showing phylogenetic distribution of subgingival microbiota, LEfSe: linear discriminant analysis effect size

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