Evaluation of Combined Ciprofloxacin and azithromycin free and nano formulations to control biofilm producing Pseudomonas aeruginosa isolated from burn wounds

Abstract

Background: Nanoparticles are becoming increasingly important against resistant superbugs including Pseudomonas aeruginosa infections.

Aims: Exploration of Azithromycin as an adjunctive therapy to Ciprofloxacin for treatment of P. aeruginosa infections. Also, preparation of Ciprofloxacin-Azithromycin nanoparticles on chitosan nanocarrier (Cipro-AZM-CS) and assessment of its antimicrobial effect in vitro and in vivo.

Methods: Detection of biofilm production and biofilm-specific antibiotic resistance ndvB and tssC1 genes was attempted. Minimal inhibitory concentration (MIC) and Minimum biofilm eradication concentration (MBEC) were done in vitro for assessment of P. aeruginosa planktonic and biofilm forms eradication, respectively. In In vivo study, Cipro-AZM-CS and free form were used to evaluate survival rate, wound contraction and bacterial load in mice after third degree burn.

Results: All isolates were positive for biofilm production and ndvB and tssC1 genes. Majority of isolates (37, 74%) were extensively drug resistant. In the planktonic state, MIC values of Cipro-AZM free and CS forms were significantly lower than free Cipro MIC (P = 0.015 and P < 0.001 respectively). Also, Cipro-AZM free and CS MBEC values were significantly lower than that of free Cipro (P < 0.010 and P < 0.001 respectively). Furthermore, The MIC and MBEC values of free Cipro-AZM decreased significantly when challenged with Cipro-AZM-CS (P = 0.009 and P < 0.001 respectively). In vivo study combined free and Cipro-AZM-CS treated subgroups showed 100% mice survival with early resolution of infection and wound contraction (75%, 77.5% respectively) VS 45% for Cipro CS (P < 0.001).

Conclusion: Combined free and Cipro-AZM-CS showed promising results in vitro and in vivo overcoming high resistance of biofilm producing P. aeruginosa.

Keywords: Azithromycin; Biofilm; Chitosan nanocarrier; Ciprofloxacin; In vivo model; Pseudomonas aeruginosa.