Exploring the Potential Applications of Limonene in Bone Trauma Therapy: Antibacterial, Anti‐inflammatory, and Bone Regeneration Properties

Bone trauma presents a complex clinical challenge requiring simultaneous management of inflammation, infection risk, and impaired regeneration. Conventional therapies often address these issues separately, highlighting the need for multifunctional agents capable of integrated therapeutic action. Limonene, a natural monoterpene, has demonstrated broad-spectrum bioactivity, but its potential in bone trauma therapy remains underexplored. This study evaluates limonene's triple therapeutic properties: (1) Antibacterial efficacy against Escherichia coli and Staphylococcus aureus; (2) Anti-inflammatory effects on RAW 264.7 macrophages under lipopolysaccharide stimulation; and (3) Osteogenic potential in MC3T3-E1 osteoblasts by assessing proliferation and osteogenic gene expression. Limonene demonstrated concentration-dependent antibacterial efficacy, with significant growth inhibition observed against both Gram-positive and Gram-negative pathogens. Meanwhile, limonene exhibited potent anti-inflammatory activity in RAW 264.7 macrophages, as evidenced by the dose-dependent suppression of key pro-inflammatory mediators, including interleukin-1β and inducible nitric oxide synthase. Notably, in MC3T3-E1 osteoblasts, limonene significantly upregulated the expression of collagen alpha1 and enhanced osteocalcin production at optimal concentrations, suggesting its dual role in mitigating inflammation while promoting osteogenic differentiation. Collectively, these findings highlight limonene as a multifunctional therapeutic candidate for bone trauma repair, meriting further investigation through in vivo studies to validate its translational potential.