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<p>Toothbrushing is a fundamental oral hygiene behavior associated with differences in the composition of salivary microbiome, yet whether its relationship with microbial profiles differs across oral conditions remains unclear. In this study, 300 participants were initially enrolled, including healthy controls and patients with dental caries or dental fluorosis (100 per group). After exclusion of one fluorosis sample that failed sequencing, 299 unstimulated saliva samples were analyzed. Based on questionnaire data, participants were classified as high-frequency (≥ 2 times/day) or low-frequency (&lt; 2 times/day) brushers. Salivary microbiota were characterized using 16&#xa0;S rRNA gene sequencing, and microbial diversity and taxonomic composition were compared both across oral conditions within each brushing cohort and between brushing-frequency subgroups within each condition. No significant differences in α- or β-diversity were observed between brushing-frequency subgroups in the healthy or fluorosis groups. In contrast, among patients with dental caries, high-frequency brushing was associated with greater α-diversity, distinct β-diversity, and characteristic taxonomic shifts, with nine genus-level markers identified by random forest analysis (AUC = 0.7688); at the species level, <i>Serratia marcescens</i>, <i>Mobiluncus mulieris</i>, and <i>Neisseria bacilliformis</i> were significantly enriched in the high-frequency brushing subgroup, showing higher mean relative abundance and detection rates than in the low-frequency brushing subgroup. Across oral conditions, clear microbial separation was observed only in the low-frequency brushing cohort. These findings suggest that the association between self-reported toothbrushing frequency and the salivary microbiome is oral condition-dependent and most evident in dental caries.</p>

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Associations of self-reported toothbrushing frequency with salivary microbiome in caries, fluorosis, and healthy subjects

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月20日

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  • Liang Liu,
  • Qinglong Wang,
  • Yifan Zhang,
  • Yingying Zhai,
  • Shanshan Liu,
  • Weihui Chen

摘要

Toothbrushing is a fundamental oral hygiene behavior associated with differences in the composition of salivary microbiome, yet whether its relationship with microbial profiles differs across oral conditions remains unclear. In this study, 300 participants were initially enrolled, including healthy controls and patients with dental caries or dental fluorosis (100 per group). After exclusion of one fluorosis sample that failed sequencing, 299 unstimulated saliva samples were analyzed. Based on questionnaire data, participants were classified as high-frequency (≥ 2 times/day) or low-frequency (< 2 times/day) brushers. Salivary microbiota were characterized using 16 S rRNA gene sequencing, and microbial diversity and taxonomic composition were compared both across oral conditions within each brushing cohort and between brushing-frequency subgroups within each condition. No significant differences in α- or β-diversity were observed between brushing-frequency subgroups in the healthy or fluorosis groups. In contrast, among patients with dental caries, high-frequency brushing was associated with greater α-diversity, distinct β-diversity, and characteristic taxonomic shifts, with nine genus-level markers identified by random forest analysis (AUC = 0.7688); at the species level, Serratia marcescens, Mobiluncus mulieris, and Neisseria bacilliformis were significantly enriched in the high-frequency brushing subgroup, showing higher mean relative abundance and detection rates than in the low-frequency brushing subgroup. Across oral conditions, clear microbial separation was observed only in the low-frequency brushing cohort. These findings suggest that the association between self-reported toothbrushing frequency and the salivary microbiome is oral condition-dependent and most evident in dental caries.