Investigation of the Protective Effects of Graviola (Annona muricata), Moringa (Moringa oleifera Lam.) and Lactobacillus gasseri in a Pseudomonas aeruginosa Fibroblast Wound Model

Objective: This study aimed to investigate the antimicrobial, anti-inflammatory, antioxidant, and anti-apoptotic effects of Graviola (Annona muricata), Moringa oleifera, and Lactobacillus gasseri, alone and in combination, against Pseudomonas aeruginosa in vitro wound infection model. Methods: Antibacterial activity tests against Pseudomonas aeruginosa was determined using the Kirby-Bauer disk diffusion method, MIC/MBC, and FIC methods. Biofilm inhibition was measured by crystal violet staining. A scratch wound model using fibroblast cell line was infected with Pseudomonas aeruginosa. Following treatment with the extracts, cell viability, oxidative stress, cytokine levels (IL-1beta, IL-10 via ELISA), and apoptotic gene expression (BAX, BCL-2 via qPCR) were evaluated. Results: The triple combination (Graviola + Moringa + Lactobacillus gasseri) showed the strongest antibacterial activity (23 mm zone; FIC = 0.25) and notable antibiofilm effects. In infected fibroblasts, cell viability increased to with combination treatment, approaching control levels. TOS levels decreased significantly while TAC increased, indicating reduced oxidative stress. IL-1beta levels were significantly lowered, and IL-10 levels were restored. Additionally, BAX gene expression decreased by 47%, and BCL-2 increased by 50% in the triple treatment group, suggesting protection against apoptosis. Conclusion: The combined application of Graviola, Moringa oleifera, and Lactobacillus gasseri exerts synergistic antimicrobial, anti-inflammatory, antioxidant, and anti-apoptotic effects against Pseudomonas aeruginosa induced wound infection. These results will shed light on the search for alternative natural therapeutic strategies, particularly in the treatment of chronic wounds associated with antibiotic-resistant pathogens.