新型季铵盐纳米颗粒对金黄色葡萄球菌和铜绿假单胞菌生物膜抑制作用的研究

黄书满; 赵玉林; 董栋; 张亚倩; 耿静

doi:10.13201/j.issn.1001-1781.2019.04.010

新型季铵盐纳米颗粒对金黄色葡萄球菌和铜绿假单胞菌生物膜抑制作用的研究

- 郑州大学第一附属医院鼻科(郑州, 450052)

详细信息

通讯作者: 赵玉林,E-mail:zhaoyulinmail@163.com

中图分类号: R765

收稿日期: 2018-11-16

A novel nanoparticle in treatment of staphylococcus aureus and pseudomonas aeruginosa biofilms

- Department of Rhinology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China

More Information

Corresponding author: ZHAO Yulin, E-mail: zhaoyulinmail@163.com

Received Date: 16 November 2018

摘要

摘要: 目的:探讨低浓度氯化十六烷基吡啶(CPC)季铵盐纳米颗粒在体外对金黄色葡萄球菌和铜绿假单胞菌生物膜的抑制作用。方法:建立金黄色葡萄球菌ATCC25923和铜绿假单胞菌ATCC15692体外生物膜模型。制备低浓度(0.010%、0.025%和0.050%)CPC-季铵盐纳米颗粒和CPC胶体溶液。用alamarBlue法分别在CPC作用5 min和2 h后检测金黄色葡萄球菌和铜绿假单胞菌游离及其生物膜的活性。应用激光扫描共聚焦显微镜(CLSM)观察CPC-季铵盐纳米颗粒与金黄色葡萄球菌和铜绿假单胞菌生物膜之间的相互作用。结果:低浓度(0.010%、0.025%和0.050%)CPC-季铵盐纳米颗粒和CPC胶体溶液对游离金黄色葡萄球菌和铜绿假单胞菌均有显著的抗菌作用(P<0.05),对金黄色葡萄球菌和铜绿假单胞菌生物膜分别作用5 min和2 h后同样具有显著的抗菌效果(P<0.05)。在CLSM研究中,金黄色葡萄球菌生物膜的面积明显减小,铜绿假单胞菌生物膜中的死菌明显增加。结论:低浓度的CPC-季铵盐纳米颗粒即可对金黄色葡萄球菌和铜绿假单胞菌游离菌及生物膜产生显著的杀伤作用,有望应用于慢性鼻窦炎的治疗。
- 季铵盐纳米颗粒 /
- 金黄色葡萄球菌 /
- 铜绿假单胞菌 /
- 细菌生物膜
Abstract: Objective: To investigate CPC-nanoparticles of low concentrations in treatment of staphylococcus aureus and pseudomonas aeruginosa biofilms in vitro. Method: We established specific biofilms of staphylococcus aureus ATCC 25923 and pseudomonas aeruginosa ATCC 15692, and prepared CPC-nanoparticles and CPC micelle solutions of low concentrations(0.010%, 0.025% and 0.050%). AlamarBlue was used to test the viability of both planktonic staphylococcus aureus and pseudomonas aeruginosa and their biofilms after treatment for 5 minutes and 2 hours respectively in the bactericidal efficacy study.The interaction between CPC-nanoparticles and staphylococcus aureus and pseudomonas aeruginosa biofilms was observed by confocal laser scanning microscope(CLSM). Result: 0.010%, 0.025% and 0.050% CPC-nanoparticles and CPC micelle solutions had significant bactericidal effect on planktonic staphylococcus aureus and pseudomonas aeruginosa after five-minute exposure(P<0.05), and staphylococcus aureus and pseudomonas aeruginosa biofilms after both five-minute and two-hour treatments(P<0.05). In CLSM study, the size of staphylococcus aureus biofilms decreased, while dead bacteria of pseudomonas aeruginosa biofilms increased after two-hour treatment.Conclusion: CPC-nanoparticles had significant bactericidal effects on staphylococcus aureus and pseudomonas aeruginosa biofilms, which could be used in treatment of CRS.
- nanoparticle /
- staphylococcus aureus /
- pseudomonas aeruginosa /
- bacterial biofilm

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参考文献(26)

[1]	] BOASE S,FOREMAN A,CLELAND E,et al.The microbiome of chronic rhinosinusitis:culture,molecular diagnostics and biofilm detection[J].BMC Infect Dis,2013,13:210.
[2]	DONG D,THOMAS N,THIERRY B,et al.Distribution and inhibition of liposomes on staphylococcus aureus and pseudomonas aeruginosa biofilm[J].PLoS One,2015,10:e0131806.
[3]	SINGHAL D,FOREMAN A,JERVIS-BARDY J,et al.staphylococcus aureus biofilms:nemesis of endoscopic sinus surgery[J].Laryngoscope,2011,121:1578-1583.
[4]	BENDOUAH Z,BARBEAU J,HAMAD W A,et al.Biofilm formation by Staphylococcus aureus and Pseudomonas aeruginosa is associated with an unfavorable evolution after surgery for chronic sinusitis and nasal polyposis[J].Otolaryngol Head Neck Surg,2006,134:991-996.
[5]	BESSA L J,EATON P,DEMATEI A,et al.Synergistic and antibiofilm properties of ocellatin peptides against multidrug-resistant Pseudomonas aeruginosa[J].Future Microbiol,2018,13:151-163.
[6]	ZIMLICHMAN E,HENDERSON D,TAMIR O,et al.Health Care-Associated Infections:A Meta-Analysis of Costs and Financial Impact on the US Health Care System[J].JAMA Int Med,2013,173:2039-2046.
[7]	FERRER-TASIES L,MORENO-CALVO E,CANO-SARABIA M,et al.Quatsomes: vesicles formed by self-assembly of sterols and quaternary ammonium surfactants[J].Langmuir,2013,29:6519-6528.
[8]	COTTENYE N,CUI Z K,WILKINSON K J,et al.Interactions between non-phospholipid liposomes containing cetylpyridinium chloride and biofilms of Streptococcus mutans:modulation of the adhesion and of the biodistribution[J].Biofouling,2013,29:817-827.
[9]	MCDONNELL G,RUSSELL A D.Antiseptics and disinfectants:activity,action,and resistance[J].Clin Microbiol Rev,1999,12:147-179.
[10]	GILBERT P,MOORE L E.Cationic antiseptics:diversity of action under a common epithet[J].J Appl Microbial,2005,99:703-715.
[11]	SOUMET C,FOURREAU E,LEGRANDOIS P,et al.Resistance to phenicol compounds following adaptation to quaternary ammonium compounds in Escherichia coli[J].Vet Microbiol,2012,158:147-152.
[12]	FROMM-DORNIEDEN C,REMBE J D,SCHÄFER N,et al.Cetylpyridinium chloride and miramistin as antiseptic substances in chronic wound management-prospects and limitations[J].J Med Microbiol,2015,64:407-414.
[13]	THOMAS N,DONG D,RICHTER K,et al.Quatsomes for the treatment of Staphylococcus aureus biofilm[J].J Materials Chemistry B,2014,11:001953.
[14]	CUI Z K,BOUISSE A,COTTENYE N,et al.Formation of pH-sensitive cationic liposomes from a binary mixture of monoalkylated primary amine and cholesterol[J].Langmuir,2012,28:13668-13674.
[15]	AHMED K,GRIBBON P N,JONES M N.The application of confocal microscopy to the study of liposome adsorption onto bacterial biofilms[J].J Liposome Res,2002,12:285-300.
[16]	PEETERS E,NELIS H J,COENYE T.Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates[J].J Microbiol Methods,2008,72:157-165.
[17]	AL-MUTAIRI D,KILTY S J.Bacterial biofilms and the pathophysiology of chronic rhinosinusitis[J].Curr Opin Allergy Clin Immunol,2011,11:18-23.
[18]	FOREMAN A,BOASE S,PSALTIS A,et al.Role of bacterial and fungal biofilms in chronic rhinosinusitis[J].Curr Allergy Asthma Rep,2012,12:127-135.
[19]	MAJIDPOUR A,FATHIZADEH S,AFSHAR M,et al.Dose-dependent effects of common antibiotics used to treat staphylococcus aureus on biofilm formation[J].Iran J Pathol,2017,12:362-370.
[20]	MARCOTTE L,THERIEN-AUBIN H,SANDT C,et al.Solute size effects on the diffusion in biofilms of Streptococcus mutans[J].Biofouling,2004,20:189-201.
[21]	GUIOT E,GEORGES P,BRUN A,et al.Heterogeneity of diffusion inside microbial biofilms determined by fluorescence correlation spectroscopy under two-photon excitation[J].Photochem Photobiol,2002,75:570-578.
[22]	BERESWILL S,VEY T,KIST M.Susceptibility in vitro of Helicobacter pylori to cetylpyridinium chloride[J].FEMS Immunol Med Microbiol,1999,24:189-192.
[23]	FLEMMING H C,WINGENDER J.The biofilm matrix[J].Nat Rev Microbiol,2010,8:623-633.
[24]	FLEMMING H C,NEU T R,WOZNIAK D J.The EPS matrix:the "house of biofilm cells"[J].J Bacteriol,2007,189:7945-7947.
[25]	BJARNSHOLT T,ALHEDE M,ALHEDE M,et al.The in vivo biofilm[J].Trends Microbial,2013,21:466-474.
[26]	THET N T,WALLACE L,WIBAUX A,et al.Development of a mixed-species biofilm model and its virulence implications in device related infections[J].J Biomed Mater Res B Appl Biomater,2018,[Epub ahead of print]