Fusion between fluid liposomes and intact bacteria: study of driving parameters and in vitro bactericidal efficacy

Zhao Wang,1,2* Yufan Ma,1,3,4* Hayssam Khalil,1 Rutao Wang,1–3 Tingli Lu,1 Wen Zhao,1 Yang Zhang,3 Jamin Chen,1,2 Tao Chen,1–3  1Key Laboratory for Space Bioscience and Biotechnology, School of Life Sciences, Northwestern Polytechnical University, Xi'an, Shaanxi,...

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Autores principales:	Wang Z, Ma Y, Khalil H, Wang R, Lu T, Zhao W, Zhang Y, Chen J, Chen T
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2016
Materias:	fusion lipid-mixing assay Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/d5a4896a7e6246a082d1a1184a59e118
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spelling	oai:doaj.org-article:d5a4896a7e6246a082d1a1184a59e1182021-12-02T02:10:25ZFusion between fluid liposomes and intact bacteria: study of driving parameters and in vitro bactericidal efficacy1178-2013https://doaj.org/article/d5a4896a7e6246a082d1a1184a59e1182016-08-01T00:00:00Zhttps://www.dovepress.com/fusion-between-fluid-liposomes-and-intact-bacteria-study-of-driving-pa-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Zhao Wang,1,2* Yufan Ma,1,3,4* Hayssam Khalil,1 Rutao Wang,1–3 Tingli Lu,1 Wen Zhao,1 Yang Zhang,3 Jamin Chen,1,2 Tao Chen,1–3  1Key Laboratory for Space Bioscience and Biotechnology, School of Life Sciences, Northwestern Polytechnical University, Xi'an, Shaanxi, 2Shaanxi Liposome Research Center, Xi'an, Shaanxi, 3Xi'an Libang Pharmaceuticals Co, Ltd, Xi'an, 4School of Medicine, Xi'an Jiaotong University, Xi'an, People's Republic of China *These authors contributed equally to this work. Background: Pseudomonas aeruginosa represents a good model of antibiotic resistance. These organisms have an outer membrane with a low level of permeability to drugs that is often combined with multidrug efflux pumps, enzymatic inactivation of the drug, or alteration of its molecular target. The acute and growing problem of antibiotic resistance of bacteria to conventional antibiotics made it imperative to develop new liposome formulations for antibiotics, and investigate the fusion between liposome and bacterium. Methods: In this study, the factors involved in fluid liposome interaction with bacteria have been investigated. We also demonstrated a mechanism of fusion between liposomes (1,2-dipalmitoyl-sn-glycero-3-phosphocholine [DPPC]/dimyristoylphosphatidylglycerol [DMPG] 9:1, mol/mol) in a fluid state, and intact bacterial cells, by lipid mixing assay. Results: The observed fusion process is shown to be mainly dependent on several key factors. Perturbation of liposome fluidity by addition of cholesterol dramatically decreased the degree of fusion with P. aeruginosa from 44% to 5%. It was observed that fusion between fluid liposomes and bacteria and also the bactericidal activities were strongly dependent upon the properties of the bacteria themselves. The level of fusion detected when fluid liposomes were mixed with Escherichia coli (66%) or P. aeruginosa (44%) seems to be correlated to their outer membrane phosphatidylethanolamine (PE) phospholipids composition (91% and 71%, respectively). Divalent cations increased the degree of fusion in the sequence Fe2+ > Mg2+ > Ca2+ > Ba2+ whereas temperatures lower than the phase transition temperature of DPPC/DMPG (9:1) vesicles decreased their fusion capacity. Acidic as well as basic pHs conferred higher degrees of fusion (54% and 45%, respectively) when compared to neutral pH (35%). Conclusion: Based on the results of this study, a possible mechanism involving cationic bridging between bacterial negatively charged lipopolysaccharide and fluid liposomes DMPG phospholipids was outlined. Furthermore, the fluid liposomal-encapsulated tobramycin was prepared, and the in vitro bactericidal effects were also investigated. Keywords: fusion, lipid-mixing assay, lipid compositionWang ZMa YKhalil HWang RLu TZhao WZhang YChen JChen TDove Medical Pressarticlefusionlipid-mixing assayMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 11, Pp 4025-4036 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	fusion lipid-mixing assay Medicine (General) R5-920
spellingShingle	fusion lipid-mixing assay Medicine (General) R5-920 Wang Z Ma Y Khalil H Wang R Lu T Zhao W Zhang Y Chen J Chen T Fusion between fluid liposomes and intact bacteria: study of driving parameters and in vitro bactericidal efficacy
description	Zhao Wang,1,2* Yufan Ma,1,3,4* Hayssam Khalil,1 Rutao Wang,1–3 Tingli Lu,1 Wen Zhao,1 Yang Zhang,3 Jamin Chen,1,2 Tao Chen,1–3  1Key Laboratory for Space Bioscience and Biotechnology, School of Life Sciences, Northwestern Polytechnical University, Xi'an, Shaanxi, 2Shaanxi Liposome Research Center, Xi'an, Shaanxi, 3Xi'an Libang Pharmaceuticals Co, Ltd, Xi'an, 4School of Medicine, Xi'an Jiaotong University, Xi'an, People's Republic of China *These authors contributed equally to this work. Background: Pseudomonas aeruginosa represents a good model of antibiotic resistance. These organisms have an outer membrane with a low level of permeability to drugs that is often combined with multidrug efflux pumps, enzymatic inactivation of the drug, or alteration of its molecular target. The acute and growing problem of antibiotic resistance of bacteria to conventional antibiotics made it imperative to develop new liposome formulations for antibiotics, and investigate the fusion between liposome and bacterium. Methods: In this study, the factors involved in fluid liposome interaction with bacteria have been investigated. We also demonstrated a mechanism of fusion between liposomes (1,2-dipalmitoyl-sn-glycero-3-phosphocholine [DPPC]/dimyristoylphosphatidylglycerol [DMPG] 9:1, mol/mol) in a fluid state, and intact bacterial cells, by lipid mixing assay. Results: The observed fusion process is shown to be mainly dependent on several key factors. Perturbation of liposome fluidity by addition of cholesterol dramatically decreased the degree of fusion with P. aeruginosa from 44% to 5%. It was observed that fusion between fluid liposomes and bacteria and also the bactericidal activities were strongly dependent upon the properties of the bacteria themselves. The level of fusion detected when fluid liposomes were mixed with Escherichia coli (66%) or P. aeruginosa (44%) seems to be correlated to their outer membrane phosphatidylethanolamine (PE) phospholipids composition (91% and 71%, respectively). Divalent cations increased the degree of fusion in the sequence Fe2+ > Mg2+ > Ca2+ > Ba2+ whereas temperatures lower than the phase transition temperature of DPPC/DMPG (9:1) vesicles decreased their fusion capacity. Acidic as well as basic pHs conferred higher degrees of fusion (54% and 45%, respectively) when compared to neutral pH (35%). Conclusion: Based on the results of this study, a possible mechanism involving cationic bridging between bacterial negatively charged lipopolysaccharide and fluid liposomes DMPG phospholipids was outlined. Furthermore, the fluid liposomal-encapsulated tobramycin was prepared, and the in vitro bactericidal effects were also investigated. Keywords: fusion, lipid-mixing assay, lipid composition
format	article
author	Wang Z Ma Y Khalil H Wang R Lu T Zhao W Zhang Y Chen J Chen T
author_facet	Wang Z Ma Y Khalil H Wang R Lu T Zhao W Zhang Y Chen J Chen T
author_sort	Wang Z
title	Fusion between fluid liposomes and intact bacteria: study of driving parameters and in vitro bactericidal efficacy
title_short	Fusion between fluid liposomes and intact bacteria: study of driving parameters and in vitro bactericidal efficacy
title_full	Fusion between fluid liposomes and intact bacteria: study of driving parameters and in vitro bactericidal efficacy
title_fullStr	Fusion between fluid liposomes and intact bacteria: study of driving parameters and in vitro bactericidal efficacy
title_full_unstemmed	Fusion between fluid liposomes and intact bacteria: study of driving parameters and in vitro bactericidal efficacy
title_sort	fusion between fluid liposomes and intact bacteria: study of driving parameters and in vitro bactericidal efficacy
publisher	Dove Medical Press
publishDate	2016
url	https://doaj.org/article/d5a4896a7e6246a082d1a1184a59e118
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Fusion between fluid liposomes and intact bacteria: study of driving parameters and in vitro bactericidal efficacy

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