DNase-I functionalization of ciprofloxacin-loaded chitosan nanoparticles overcomes the biofilm-mediated resistance of Pseudomonas aeruginosa

Pseudomonas aeruginosa infection in cystic fibrosis (CF) is a major complication which aggravates the disease complexity and progression. The improvement of antimicrobial therapy against the P. aeruginosa biofilm infection in CF by dissembling the extracellular matrix is a fundamental concept of this study. Chitosan nanoparticle was successfully fabricated and characterized. Furthermore, various in vitro antimicrobial efficacy, biofilm dispersal potential, and in vitro as well as in vivo toxicity were assessed. The chitosan nanoparticles of ciprofloxacin functionalized with DNase-I had spherical shape with desired quality attributes (particle size-212.3 +/- 8.9; polydispersity index-0.288 +/- 0.06, zeta potential-14.6 +/- 1.3, and entrapment efficiency-53.7 +/- 3.8%). The developed DNase-I functionalized chitosan nanoparticles laden with ciprofloxacin demonstrated substantial and prolonged microbial inhibition, efficiently prevented the biofilm development, and possessed the excellent biofilm dispersal potential. Moreover, the confocal study demonstrated that the biofilm treated with DNase-I functionalized chitosan nanoparticles of ciprofloxacin had minimum biofilm thickness, biomass, and microbial density compared to the other treatment groups. Additionally, the developed formulation was found to be safe and had minimal in vitro as well as in vivo toxicity. Thus, the findings of this study suggests ciprofloxacin-loaded chitosan nanoparticles functionalized with DNase-I as an effective and safe treatment approach for the P. aeruginosa infection in CF.