Restoring ciprofloxacin efficacy via β-caryophyllene synergy for concurrent infection control and tissue regeneration.

The occurrence of antimicrobial-resistant microbial infection on the wounds are considered a critical issue in delayed or non-healing wounds. Due to the scarcity of new antibiotics in the market, alternate combinational therapies are gaining interest in tackling this issue. In this study, we investigate the antimicrobial efficacy of ciprofloxacin (CIP) enhanced by the adjuvant activity of beta-caryophyllene (BCP), an efflux pump inhibitor harnessing their synergistic potential to achieve improved wound healing activity. The synergistic effect of CIP and BCP was assessed for its antimicrobial activity and evaluated for wound healing and infection control in both in vitro and in ovo models. The non-cytotoxic concentration of BCP (80 µg/mL) and varying concentrations of CIP (1.25-10 µg/mL) combination effectively reduced CIP's minimum inhibitory concentration by half and disintegrated biofilms of Staphylococcus aureus and Enterococcus faecalis. The combination increased fibroblast cell migration by 20% in in vitro compared to untreated samples. The BCP treatment modulated key wound-healing-associated genes, with upregulation of TGF-β, COL1A1, COL3A1, and FGF2, and downregulation of IL-1β and MMP-2, indicating profound wound healing activity. In the in ovo infection model, it achieved a 100% survival rate and additionally led to a 45% increase in vessel length and a 60% rise in vessel junctions in angiogenesis studies. Therefore, the combinational therapy of CIP and BCP demonstrated a dual role by effectively managing infections caused by multidrug-resistant wound pathogens and promoting wound healing. Remarkably, this approach reduced CIP dosage by 50%, thereby lowering antibiotic selective pressure and yielded complete killing of pathogens.