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Multi-functional Liposomes Enhancing Target and Antibacterial Immunity for Antimicrobial and Anti-Biofilm Against Methicillin-Resistant Staphylococcus aureus

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Abstract

Purpose

The aim of this study was to prepare wheat germ agglutinin (WGA)-modified liposomes encapsulating clarithromycin and to evaluate their in vitro and in vivo efficacy against Methicillin-resistant Staphylococcus aureus (MRSA).

Methods

Physicochemical parameters, minimum inhibitory concentrations, in vitro killing kinetic, cellular uptake, biofilm formation inhibition and pre-formed biofilm destruction, biodistribution, in vivo antibacterial efficacy against MRSA, and phagocytosis into macrophages for liposomes loading clarithromycin were determined.

Results

The minimum inhibitory concentration and the time–kill curve for WGA-modified liposomal clarithromycin were better than those of free and nonmodified liposomal clarithromycin. Flow cytometry analysis displayed that liposomes could deliver more Coumarin 6, a fluorescent probe, into bacteria because of the conjugation of WGA. Besides, WGA-modified liposomal clarithromycin inhibited formation of S. aureus (ATCC 29213) and MRSA biofiom, and prompted the biofilm disassembly at lower concentrations below MIC. Effective accumulation of liposomes was displayed in the enterocoelia of the mice because of WGA. The number of MRSA colony-forming units in the kidney and spleen in mice treated with WGA-modified liposomal clarithromycin was significantly lower than that treated with free and nonmodified clarithromycin (p < 0.05). Intracellular localization of MRSA occurred in a significantly higher proportion of macrophage exposed to WGA-modified liposomes compared to those exposed to nonmodified liposomes.

Conclusions

Liposome modified by WGA is a promising formulation for bacteria targeted delivery and immunity defensive system through macrophage improving uptake of bacteria, biodistribution, in vitro and in vivo antibacterial efficacy against MRSA.

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Abbreviations

CLSM:

Confocal laser scanning microscope

DC-Chol:

Cholesteryl 3β-N-di-methyl-amino-ethyl-carbamate hydrochloride

DiR:

DiR iodide [1,1′-dioctadecyl-3,3,3′,3′-tetramethylindotricarbocyanine iodide]

DMEM:

Dulbecco minimum essential medium

DSPE-PEG:

DSPE-PEG2000

Lip:

Clarithromycin loaded liposomes

MIC:

Minimum inhibitory concentration

MRSA:

Methicillin-resistant staphylococcus aureus

NHS-PEG-DSPE:

N-hydroxysuccinimidyl-PEG2000-DSPE

PBS:

Phosphate-buffered saline

PDI:

Polydispersity index

PEG-Lip:

Nonmodified liposomes

S. aureus :

Staphylococcus aureus

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

SEM:

Scanning Electron Microscopy

SPC:

Soy phosphatidylcholine

TSA:

Tryptic soy agar

TSB:

Tryptic soy broth

WGA:

Wheat germ agglutinin

WGA-Lip:

WGA-modified liposomes

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ACKNOWLEDGMENTS AND DISCLOSURES

This work was supported by the National Natural Science Foundation of China (No. 81202488).

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Author notes

    Authors and Affiliations

    1. Department of Pharmaceutics, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China

      Yansha Meng, Jiongxi Lei, Mengmeng Chen, Shuangchen Cong, Yuanyuan Zhang & Xinru Li

    2. Institute of Medicinal Biotechnology, Chinese Academy of Medical Science & Peking Union Medical College, Beijing, 100050, China

      Xucheng Hou, Weiming Ding & Guiling Li

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    1. Yansha Meng
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    2. Xucheng Hou
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    3. Jiongxi Lei
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    Correspondence to Guiling Li or Xinru Li.

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    Yansha Meng and Xucheng Hou contributed equally to this work.

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    Meng, Y., Hou, X., Lei, J. et al. Multi-functional Liposomes Enhancing Target and Antibacterial Immunity for Antimicrobial and Anti-Biofilm Against Methicillin-Resistant Staphylococcus aureus . Pharm Res 33, 763–775 (2016). https://doi.org/10.1007/s11095-015-1825-9

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    • DOI: https://doi.org/10.1007/s11095-015-1825-9

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