Biofilm Eradication Four-Step Strategy: Study of Using Self-Assembled Azithromycin/Rhamnolipid Nanoparticles for Removing <i>Pseudomonas aeruginosa</i> Biofilm

DONG Ya-ting; LI Peng-yu; SUN Ying-ying; RAO Yi-qin; YU Shi-hui; HU Hai-yan

doi:10.12182/20210760207

Volume 52 Issue 4

Jul. 2021

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JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCES) > 2021 > 52(4): 598-604. > DOI: 10.12182/20210760207

DONG Ya-ting, LI Peng-yu, SUN Ying-ying, et al. Biofilm Eradication Four-Step Strategy: Study of Using Self-Assembled Azithromycin/Rhamnolipid Nanoparticles for Removing Pseudomonas aeruginosa Biofilm[J]. Journal of Sichuan University (Medical Sciences), 2021, 52(4): 598-604. DOI: 10.12182/20210760207

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Biofilm Eradication Four-Step Strategy: Study of Using Self-Assembled Azithromycin/Rhamnolipid Nanoparticles for Removing Pseudomonas aeruginosa Biofilm

School of Pharmaceutical Science, Sun Yat-Sen University, Guangzhou 510006, China

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Corresponding author:
HU Hai-yan, E-mail：lsshhy@mail.sysu.edu.cn
Received Date: March 21, 2021
Revised Date: May 31, 2021
Available Online: July 21, 2021
Published Date: July 19, 2021

Abstract

Abstract

Objective To investigate the in vitro eradicative effect of self-assembled azithromycin/rhamnolipid nanoparticles (AZI-RHL NPs) on Pseudomonas aeruginosa (P. aeruginosa) biofilm.
Methods AZI-RHL NPs were prepared and characterized. The minimum inhibitory concentration (MIC) of AZI-RHL NPs on planktonic P. aeruginosa was measured by the broth microdilution method. The eradicative effect of AZI-RHL NPs on P. aeruginosa biofilm was evaluated via crystal violet staining and SYTO 9/PI live/dead staining. Fluorescence labeling was used to measure the eradicative effect of NPs on extracellular polymeric substances (EPS). In addition, crystal violet staining was performed to evaluate the inhibitory effect of AZI-RHL NPs on the adhesion of P. aeruginosa on human bronchial epithelial BEAS-2B cells. To investigate the ability of AZI-RHL NPs to penetrate mucus, the interaction between NPs and mucin was measured via particle size changes after co-incubation with mucin solution.
Results The AZI-RHL NPs had a particle size of about 121 nm and were negatively charged on the surface, displaying a high encapsulation efficiency and a high drug loading capacity of 96.72% and 45.08% for AZI, respectively and 99.38% and 53.07% for RHL, respectively. The MIC of AZI-RHL NPs on planktonic P. aeruginosa was half of that of using AZI alone. AZI-RHL NPs displayed the capacity to effectively destroy the biofilm structure and remove the proteins and polysaccharides in EPS, eradicating biofilms in addition to reducing the survival rate of bacteria in the biofilm. AZI-RHL NPs were shown to have inhibited P. aeruginosa adhesion on BEAS-2B cells and prevented the residual bacteria from forming a new biofilm. There was no significant change in the particle size of NPs after co-incubation with mucin solution, indicating a weak interaction between NPs and mucin, and suggesting that NPs could penetrate the mucus and reach the P. aeruginosa infection sites.
Conclusion AZI-RHL NPs were able to effectively enhance the removal of P. aeruginosa biofilm through a four-step strategy of biofilm eradication, including penetrating the mucus, disintegrating the biofilm structure, killing the bacteria dispersed from biofilm, and preventing the adhesion of residual bacteria. We hope that this study will provide a replicable common strategy for the treatment of refractory infections caused by P. aeruginosa and other types of biofilms.
- Azithromycin,
- Rhamnolipid,
- Self-assembled nanoparticles,
- Pseudomonas aeruginosa,
- Antibiofilm

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Biofilm Eradication Four-Step Strategy: Study of Using Self-Assembled Azithromycin/Rhamnolipid Nanoparticles for Removing Pseudomonas aeruginosa Biofilm

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