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Incorporation of farnesol significantly increases the efficacy of liposomal ciprofloxacin against Pseudomonas aeruginosa biofilms in vitro H.M.H.N. Bandara, M.J. Herpin, D. Kolacny, A. Harb, D. Romanovicz, and H.D.C. Smyth Mol. Pharmaceutics, Just Accepted Manuscript • DOI: 10.1021/acs.molpharmaceut.6b00360 • Publication Date (Web): 06 Jul 2016 Downloaded from http://pubs.acs.org on July 8, 2016
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Molecular Pharmaceutics
Incorporation of farnesol significantly increases the efficacy of liposomal ciprofloxacin against Pseudomonas aeruginosa biofilms in vitro H.M.H.N. Bandara1,3, M.J. Herpin1, D. Kolacny Jr.1, A. Harb1, D. Romanovicz2, & H.D.C. Smyth1*
1Division
of Pharmaceutics, College of Pharmacy, The University of Texas at Austin, Austin,
Texas, USA. 2Institute
of Cellular and Molecular Biology, College of Natural Sciences, The University of
Texas at Austin, Austin, Texas, USA. 3
Current address: School of Dentistry, The University of Queensland, Australia
* Corresponding author:
Dr. Hugh D. C. Smyth, College of Pharmacy, 2409 West University Avenue, PHR 4.214, Austin, TX, USA 78712 Phone: +1 (512) 471 3383 Fax: +1 (512) 471 7474 E mail:
[email protected] ACS Paragon Plus Environment
Molecular Pharmaceutics
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Abstract The challenge of eliminating Pseudomonas aeruginosa infections, such as in cystic fibrosis lungs, remains unchanged due to the rapid development of antibiotic resistance. Poor drug penetration into dense P. aeruginosa biofilms plays a vital role in ineffective clearance of the infection. Thus, the current antibiotic therapy against P. aeruginosa biofilms need to be revisited and alternative antibiofilm strategies need to be invented. Fungal quorum sensing molecule (QSM), farnesol, appear to have detrimental effects on P. aeruginosa. Thus, this study aimed to co-deliver naturally occurring QSM farnesol, with the antibiotic ciprofloxacin as a liposomal formulation to eradicate P. aeruginosa biofilms. Four different liposomes (with ciprofloxacin and farnesol: Lcip+far, with ciprofloxacin:
Lcip,
with farnesol: Lfar, control: Lcon) were prepared using dehydration-rehydration method and characterized. Drug entrapment and release were evaluated by spectrometry and high performance liquid chromatography (HPLC). The efficacy of liposomes was assessed using standard biofilm assay. Liposome-treated 24h P. aeruginosa biofilms were quantitatively assessed by XTT reduction assay and crystal violet assay, qualitatively by confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM). Ciprofloxacin release from liposomes was higher when encapsulated with farnesol (Lcip+far ) compared to Lcip (3.06% vs 1.48%) whereas farnesol release was lower when encapsulated with ciprofloxacin (Lcip+far ) compared to Lfar (1.81% vs 4.75%). The biofilm metabolism was significantly lower when treated with Lcip+far or Lcip compared to free ciprofloxacin (XTT, P
