Figure 5.

(A) Schematic approach for the preparation of gentamicin-NONOate nanoparticles via RAFT polymerization. (B and C) GEN-NO nanoparticles induced dispersal in P. aeruginosa biofilms. (B) Bacterial biofilms were grown in multi-well plates for 6 h in the absence of any treatment before being treated for a further 1 h with various concentrations (mM) of NO donor spermine NONOate (Sper-NO), free gentamicin or gentamicin-conjugated polymers (Poly-GEN) and GEN-NO nanoparticles (Poly-GENNO). Biofilm biomass was analyzed by crystal violet staining. Error bars represent standard error (n=2). (C) Stained biofilms treated with the indicated concentrations of GEN-NO nanoparticles. Note: concentration based on GEN, one mole of GEN-NO nanoparticles is equivalent to one mole of Sper-NO and gentamicin. (D) Representative confocal images showing P. aeruginosa biofilms stained with LIVE/DEAD kit. Biofilms were grown for 6 h and then treated with NO donor spermine NONOate (SperNO), free gentamicin, GEN-NO nanoparticles or left untreated for a further 1 h before staining. Viable and non-viable bacteria appear green and red, as well as those stained both green/red, respectively. Scale bar=50 mm. Note: concentration based on GEN, one mole of GEN-NO nanoparticles is equivalent to one mole of Sper-NO and gentamicin. Reproduced from Nguyen T-K, Selvanayagam R, Ho KKK, et al. Co-delivery of nitric oxide and antibiotic using polymeric nanoparticles. Chem Sci. 2016;7(2):1016–1027.⁷⁰ Creative Commons license and disclaimer available from: https://creativecommons.org/licenses/by/3.0/.