Killing effect of nanoencapsulated colistin sulfate on Pseudomonas aeruginosa from cystic fibrosis patients E. Sans-Serramitjana, E. Fusté, B. Martínez-Garriga,

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Killing effect of nanoencapsulated colistin sulfate on Pseudomonas aeruginosa from cystic fibrosis patients E. Sans-Serramitjana, E. Fusté, B. Martínez-Garriga, A. Merlos, M. Pastor, J.L. Pedraz, A. Esquisabel, D. Bachiller, T. Vinuesa, M. Viñas Journal of Cystic Fibrosis Volume 15, Issue 5, Pages 611-618 (September 2016) DOI: 10.1016/j.jcf.2015.12.005 Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

Fig. 1 Effect of free colistin, NLC of colistin, and mixtures of empty nanoparticles and free colistin on P. aeruginosa growth. (a) 056SJD at 1/4× MIC (0.5μg/mL); (b) 056SJD at MIC (2μg/mL); (c) 056SJD at 8× MIC (16μg/mL); (d) P19 at 1/4× MIC (8μg/mL); (e) P19 at MIC (32μg/mL), and (f) P19 at 8x MIC (256μg/mL). All incubations were at 37°C with shaking at 2000rpm and bacteria were in the exponential phase of growth. Journal of Cystic Fibrosis 2016 15, 611-618DOI: (10.1016/j.jcf.2015.12.005) Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

Fig. 2 AFM images obtained at 7μm2 of scan size of (a) untreated P. aeruginosa biofilms; (b) after treatment with 128μg/mL of free (first row) and nanoencapsulated (second row) colistin sulfate at 2h; (c) after treatment with 128μg/mL of free (first row) and nanoencapsulated (second row) colistin sulfate at 6h; (d) after treatment with 128μg/mL of free (first row) and nanoencapsulated (second row) colistin sulfate at 24h. First column shows topography images, whereas amplitude images are shown in second column. Journal of Cystic Fibrosis 2016 15, 611-618DOI: (10.1016/j.jcf.2015.12.005) Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

Fig. 3 Graphical representation of surface roughness (Ra) in nanometers, according to the different formulations tested and surface scans. Bars represent the standard deviation of the mean. Journal of Cystic Fibrosis 2016 15, 611-618DOI: (10.1016/j.jcf.2015.12.005) Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

Fig. 4 Typical force–distance curves of untreated and colistin and NLC-colistin-treated P. aeruginosa 056SJD. Journal of Cystic Fibrosis 2016 15, 611-618DOI: (10.1016/j.jcf.2015.12.005) Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions