Abstract
BACKGROUND: The oral microbiome is vital for health, yet population-based evidence on how self-reported flossing relates to microbial communities remains limited. This study examined the association between self-reported dental flossing frequency and oral microbiome diversity in a nationally representative sample of U.S. adults. METHODS: This cross-sectional analysis included 4,772 adults aged 30-69 from NHANES 2009-2012. Flossing frequency was categorized as non-users (0 days/week), some flossing (1-6 days/week), and daily users (7 days/week). Oral microbiome composition was profiled using 16S rRNA sequencing. α-diversity was calculated using Observed amplicon sequence variants (ASVs), Shannon, Inverse Simpson, and Faith's Phylogenetic Diversity (PD); β-diversity using Bray-Curtis and UniFrac distances. Survey-weighted linear regression and PERMANOVA were used with covariate adjustment. RESULTS: Participants included, non-users (32%), some flossing (38%), and daily users (30%). A dose-response relationship was observed between flossing frequency and reduced microbial richness and phylogenetic diversity. Compared with non-users, daily users exhibited significantly lower richness (Observed ASVs: β = -11.46, 95% CI: -15.62 to -7.29) and phylogenetic diversity (Faith's PD: β = -0.88, 95% CI: -1.20 to -0.56). Daily flossing was associated with a modest reduction in Shannon diversity, with no significant association for the Inverse Simpson index. Inverse associations were more pronounced among younger and lower-income adults, but not among current smokers. β-diversity differed significantly across groups, although effect sizes were minimal (Bray-Curtis R(2) = 0.059%; unweighted UniFrac R(2) = 0.090%). CONCLUSIONS: Frequent flossing was associated with reduced microbial richness and phylogenetic diversity, potentially indicating a favorable shift toward a healthier microbial community.