Objective:

To investigate the effects of Berberine (BBR) against Staphylococcus aureus (SA)-induced osteomyelitis (OM) and explore potential mechanisms through network pharmacology and validation.

Approach:

• In vitro and In vivo Studies: An in vitro SA biofilm model was established to assess BBR's effects on bacterial survival and biofilm structure. A mouse model of SA-induced OM was used, with a clindamycin-treated group as a positive comparator.
• Microscopic and Biochemical Analysis: Biofilm formation was examined using scanning electron microscopy (SEM), and serum inflammatory mediators were measured by ELISA. Histopathological changes were evaluated by H&E staining.
• Network Pharmacology and Molecular Docking: Network pharmacology was performed to identify targets and pathways, with key proteins validated by Western blotting.

Key Findings:

• BBR significantly reduced bacterial viability within biofilms and decreased CFU counts in vitro.
• Microscopic observations showed disrupted biofilm architecture and reduced biofilm coverage after BBR treatment.
• In vivo, BBR alleviated bone destruction, reduced implant-associated biofilm formation, and decreased inflammatory cell infiltration.
• ELISA results indicated that BBR markedly reduced pro-inflammatory mediator levels.
• Network pharmacology identified PTGS2 as a key target and implicated the HIF-1 signaling pathway.

Interpretation:

BBR exerts protective effects against SA-induced OM through antibiofilm and anti-inflammatory activities, but its antibacterial efficacy is weaker than that of clindamycin.

Limitations:

• BBR should not be regarded as a replacement for antibiotics but rather as an adjunctive therapy for biofilm-associated osteomyelitis.

Conclusion:

BBR demonstrates potential in mitigating immune-inflammatory damage in SA-induced osteomyelitis, warranting further investigation.