种植修复患者唾液与龈下菌斑微生物群落结构关联及其影响因素研究

Correlation between saliva and subgingival plaque microbial community structure and its influencing factors in patients with implant restoration

目的 利用高通量测序技术研究种植修复患者唾液与龈下菌斑微生物群落结构的关联及其影响因素,为种植体周微生物研究的样本选择提供参考。方法 选取于2014年6月至2015年9月在第四军医大学口腔医院口腔修复科完成种植修复的10例患者(12颗种植体),于2021年11月至2022年1月随访复诊时采集其唾液和种植体周龈下菌斑样本,行16S rRNA V3~V4区双端测序,通过Alpha多样性分析和LEfSe多级物种差异判别分析其微生物群落丰富度、多样性及物种丰度的差异性。结果 在Alpha多样性分析中,Sobs指数和Chao指数显示,唾液中微生物群落丰富度大于龈下菌斑,差异均有统计学意义(均P <0.05);而Shannon指数和Simpson指数显示,唾液与龈下菌斑微生物群落多样性差异无统计学意义(P> 0.05)。进一步分析发现,种植体周围黏膜炎患者和吸烟患者的唾液与龈下菌斑微生物群落丰富度差异无统计学意义(均P> 0.05);男性、不饮酒、种植体周最大探诊深度≥5 mm的患者唾液与龈下菌斑微生物群落多样性差异有统计学意义(均P <0.05);其他因素如不同年龄、软垢指数、牙石指数、角化黏膜宽度、固位方式、改良龈沟出血指数、边缘骨吸收量、牙周炎患病状况的患者唾液与龈下菌斑微生物群落丰富度差异有统计学意义(均P <0.05),而微生物群落多样性差异均无统计学意义(均P> 0.05)。LEfSe分析显示,唾液与龈下菌斑微生物组成相似,在属水平、种水平分别仅有7种和12种微生物具有差异(均P <0.05)。结论 种植修复患者唾液与龈下菌斑微生物群落结构关联密切,男性、不饮酒、种植体周最大探诊深度≥5 mm对种植修复患者唾液及龈下菌斑微生物群落关联有影响,若无此类影响因素可考虑用唾液替代种植体周围龈下菌斑进行种植体周微生物群落多样性的研究。

Objective To investigate the relationship between saliva and the subgingival plaque microbial community structure of patients with implant restorations and its influencing factors using high-throughput sequencing technology,and to serve as a guide for the sample selection of peri-implant microbiological research. Methods Saliva and peri-implant subgingival plaque samples were collected from 10 patients(12 implants)at their follow-up visit between November 2021 and September 2022,who completed implant restorations in the Department of Prosthodontics,Stomatological Hospital of the Fourth Military Medical University from June 2014 to September 2015. The double-ended 16S r RNA V3-V4 sequencing was performed. Alpha diversity analysis and LEfSe analysis were used to study the differences in microbial community richness,diversity,and species abundance. Results In the Alpha diversity analysis,Sobs and Chao index showed that the microbial community richness in saliva was greater than that in subgingival plaque,and the differences were statistically significant(P < 0.05). The Shannon and Simpson index,however,revealed that there were no significant differences between subgingival plaque and saliva in terms of the richness of microbial commu-nities(P > 0.05). Further analysis showed that there were no significant differences in the richness of microbial communities in saliva and subgingival plaque between patients with peri-implant mucositis and those who smoked(P > 0.05),while the differences in saliva and subgingival plaque community diversity were statistically significant in male,non-alcoholic patients and patients with implant maximum probing depth ≥ 5 mm(P < 0.05). Other factors such as age,debris index,calculus index,keratinized mucosal width,retention methods,modified sulcus bleeding index,marginal bone resorption volume,and periodontal health status had significant effects on the microbial community richness of saliva and subgingival plaque,and the differences were statis-tically significant,while community diversity had no statistical difference(P > 0.05). LEfSe analysis showed that the microbial compositions of saliva and sub-gingival plaque were similar,and only 7 and 12 microbial species were different at the genus and species levels,respectively(P < 0.05). Conclusion Saliva is closely correlated with the microbial community structure of the subgingival plaque in patients with implant restoration. The link between saliva and the subgingival plaque microbial community in patients with implant restoration is influenced by male,no alcohol consumption,and peri-implant maximum probing depth ≥ 5 mm. For the study of peri-implant microbial community diversity,saliva can be used to replace peri-implant subgingival plaque in the absence of these elements.