Authors:

Anna M Kiley

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The gut microbiome affects skeletal health by influencing immune signaling, nutrient absorption, and inflammation in bone remodeling. While sex-based differences are known in both gut microbiota and bone biology, it remains unknown whether the gut-bone axis itself is also sex- dependent. Understanding how microbial composition varies with antibiotic treatment, duration, and sex may reveal sex-specific strategies to treat musculoskeletal conditions. To study this, we used a mouse gut microbiome model. Under institutional approval, mice received either no treatment or antibiotics in water (1g/L ampicillin + 0.5g/L neomycin) starting at 5 weeks of age. At euthanasia after 3, 11, and 19 weeks, we collected cecum contents, serum, and hindlimbs. 16S rRNA sequencing enabled phylum-level relative abundance analysis of the gut microbiome. Using two-way ANOVA (RStudio), we assessed sex and treatment effects at each time point. After 3 weeks, antibiotics affected Actinobacteria (p = 0.005) and Bacteroidetes (p = 0.001); Bacteroidetes also showed a sex effect (p = 0.03). At 11 weeks, Bacteroidetes was influenced by sex (p = 0.01) and treatment (p = 0.003); Firmicutes differed by sex (p < 0.001). Tenericutes showed sex, treatment, and interaction effects (p = 0.01, 0.002, < 0.001). Verrucomicrobia had a treatment effect (p < 0.001). By 19 weeks, treatment effects became dominant: Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria, and Verrucomicrobia all showed treatment effects. These results suggest sex shapes early microbiome changes, while prolonged antibiotics drive sustained shifts. This may help guide future research leveraging the gut- bone axis to prevent or treat musculoskeletal disease. Keywords: Gut Microbiome; Gut-Bone Axis; Sex-Based Differences; Antibiotic Treatment

Source:

Purdue University / 2025

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Co-authors:

Anna M Kiley