Photodynamic activation of phytochemical-antibiotic combinations for combatting Staphylococcus aureus from acute wound infections

Abstract

Staphylococcus aureus is characterized by its high resistance to conventional antibiotics, particularly methicillinresistant (MRSA) strains, making it a predominant pathogen in acute and chronic wound infections. The persistence of acute S. aureus wound infections poses a threat by increasing the incidence of their chronicity. This study investigated the potential of photodynamic activation using phytochemical-antibiotic combinations to eliminate S. aureus under conditions representative of acute wound infections, aiming to mitigate the risk of chronicity. The strategy applied takes advantage of the promising antibacterial and photosensitising properties of phytochemicals, and their ability to act as antibiotic adjuvants. The antibacterial activity of selected phytochemicals (berberine, curcumin, farnesol, gallic acid, and quercetin; 6.25 - 1000 mu g/mL) and antibiotics (ciprofloxacin, tetracycline, fusidic acid, oxacillin, gentamicin, mupirocin, methicillin, and tobramycin; 0.0625 - 1024 mu g/mL) was screened individually and in combination against two S. aureus clinical strains (methicillin-resistant and-susceptible - MRSA and MSSA). The photodynamic activity of the phytochemicals was assessed using a lightemitting diode (LED) system with blue (420 nm) or UV-A (365 nm) variants, at 30 mW/cm 2 (light doses of 9, 18, 27 J/cm 2 ) and 5.5 mW/cm 2 (light doses of 1.5, 3.3 and 5.0 J/cm 2 ), respectively. Notably, all phytochemicals restored antibiotic activity, with 9 and 13 combinations exhibiting potentiating effects on MSSA and MRSA, respectively. Photodynamic activation with blue light (420 nm) resulted in an 8- to 80-fold reduction in the bactericidal concentration of berberine against MSSA and MRSA, while curcumin caused 80-fold reduction for both strains at the light dose of 18 J/cm 2 . Berberine and curcumin-antibiotic combinations when subjected to photodynamic activation (420 nm light, 10 min, 18 J/cm 2 ) reduced S. aureus culturability by approximate to 9 log CFU/mL. These combinations lowered the bactericidal concentration of antibiotics, achieving a 2048-fold reduction for gentamicin and 512-fold reduction for tobramycin. Overall, the dual approach involving antimicrobial photodynamic inactivation and selected phytochemical-antibiotic combinations demonstrated a synergistic effect, drastically reducing the culturability of S. aureus and restoring the activity of gentamicin and tobramycin.