Discrimination of Bacterial Community Structures among Healthy, Gingivitis, and Periodontitis Statuses through Integrated Metatranscriptomic and Network Analyses

ABSTRACT Periodontal disease is an inflammatory condition caused by polymicrobial infection. The inflammation is initiated at the gingiva (gingivitis) and then extends to the alveolar bone, leading to tooth loss (periodontitis). Previous studies have shown differences in bacterial composition between periodontal healthy and diseased sites. However, bacterial metabolic activities during the health-to-periodontitis microbiome shift are still inadequately understood. This study was performed to investigate the bacterial characteristics of healthy, gingivitis, and periodontitis statuses through metatranscriptomic analysis. Subgingival plaque samples of healthy, gingivitis, and periodontitis sites in the same oral cavity were collected from 21 patients. Bacterial compositions were then determined based on 16S rRNA reads; taxonomic and functional profiles derived from genes based on mRNA reads were estimated. The results showed clear differences in bacterial compositions and functional profiles between healthy and periodontitis sites. Co-occurrence networks were constructed for each group by connecting two bacterial species if their mRNA abundances were positively correlated. The clustering coefficient values were 0.536 for healthy, 0.600 for gingivitis, and 0.371 for periodontitis sites; thus, network complexity increased during gingivitis development, whereas it decreased during progression to periodontitis. Taxa, including Eubacterium nodatum, Eubacterium saphenum, Filifactor alocis, and Fretibacterium fastidiosum, showed greater transcriptional activities than those of red complex bacteria, in conjunction with disease progression. These taxa were associated with periodontal disease progression, and the health-to-periodontitis microbiome shift was accompanied by alterations in bacterial network structure and complexity. IMPORTANCE The characteristics of the periodontal microbiome influence clinical periodontal status. Gingivitis involves reversible gingival inflammation without alveolar bone resorption. In contrast, periodontitis is an irreversible disease characterized by inflammatory destruction in both soft and hard tissues. An imbalance of the microbiome is present in both gingivitis and periodontitis. However, differences in microbiomes and their functional activities in the healthy, gingivitis, and periodontitis statuses are still inadequately understood. Furthermore, some inflamed gingival statuses do not consistently cause attachment loss. In this study, metatranscriptomic analyses were used to investigate the specific bacterial composition and gene expression patterns of the microbiomes of the healthy, gingivitis, and periodontitis statuses. In addition, co-occurrence network analysis revealed that the gingivitis site included features of networks observed in both the healthy and periodontitis sites. These results provide transcriptomic evidence to support gingivitis as an intermediate state between the healthy and periodontitis statuses.