Abstract
Bacteria use a signaling mechanism called quorum sensing (QS) to form complex communities of surface-attached cells known as biofilms. This protective mode of growth allows them to resist antibiotic treatment and originates the majority of hospital-acquired infections. Emerging alternatives to control biofilm-associated infections and multidrug resistance development interfere with bacterial QS pathways, exerting less selective pressure on bacterial population. In this study, biologically stable coatings comprising the QS disrupting enzyme acylase were built on silicone urinary catheters using a layer-by-layer technique. This was achieved by the alternate deposition of negatively charged enzyme and positively charged polyethylenimine. The acylase-coated catheters efficiently quenched the QS in the biosensor strain Chromobacterium violaceum CECT 5999, demonstrated by approximately 50 % inhibition of violacein production. These enzyme multilayer coatings significantly reduced the Pseudomonas aeruginosa ATCC 10145 biofilm formation under static and dynamic conditions in an in vitro catheterized bladder model. The quorum quenching enzyme coatings did not affect the viability of the human fibroblasts (BJ-5ta) over 7 days, corresponding to the extended useful life of urinary catheters. Such enzyme-based approach could be an alternative to the conventional antibiotic treatment for prevention of biofilm-associated urinary tract infections.
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References
Blosser RS, Gray KM (2000) Extraction of violacein from Chromobacterium violaceum provides a new quantitative bioassay for N-acyl homoserine lactone autoinducers. J Microbiol Methods 40:47–55. doi:10.1016/S0167-7012(99)00136-0
Caseli L, dos Santos DS Jr, Foschini M, Gonçalves D, Oliveira ON Jr (2007) Control of catalytic activity of glucose oxidase in layer-by-layer films of chitosan and glucose oxidase. Mater Sci Eng C 27:1108–1110. doi:10.1016/j.msec.2006.08.030
Chen M, Yu Q, Sun H (2013) Novel strategies for the prevention and treatment of biofilm related infections. Int J Mol Sci 14:18488–18501. doi:10.3390/ijms140918488
Christensen LD, Moser C, Jensen PØ, Rasmussen TB, Christophersen L, Kjelleberg S, Kumar N, Høiby N, Givskov M, Bjarnsholt T (2007) Impact of Pseudomonas aeruginosa quorum sensing on biofilm persistence in an in vivo intraperitoneal foreign-body infection model. Microbiology 153:2312–2320. doi:10.1099/mic. 0.2007/006122-0
Cole SJ, Records AR, Orr MW, Linden SB, Lee VT (2014) Catheter-associated urinary tract infection by Pseudomonas aeruginosa is mediated by exopolysaccharide-independent biofilms. Infect Immun 82:2048–2058. doi:10.1128/iai. 01652-14
Darouiche RO (2004) Treatment of infections associated with surgical implants. N Engl J Med 350:1422–1429. doi:10.1056/NEJMra035415
Davies DG, Parsek MR, Pearson JP, Iglewski BH, Costerton JW, Greenberg EP (1998) The involvement of cell-to-cell signals in the development of a bacterial biofilm. Science 280:295–298. doi:10.1126/science.280.5361.295
Donlan RM (2001) Biofilm formation: a clinically relevant microbiological process. Clin Infect Dis 33:1387–1392. doi:10.1086/322972
Evliyaoǧlu Y, Kobaner M, Çelebi H, Yelsel K, Doǧan A (2011) The efficacy of a novel antibacterial hydroxyapatite nanoparticle-coated indwelling urinary catheter in preventing biofilm formation and catheter-associated urinary tract infection in rabbits. Urol Res 39:443–449. doi:10.1007/s00240-011-0379-5
Fernandes MM, Francesko A, Torrent-Burgués J, Carrión-Fité FJ, Heinze T, Tzanov T (2014) Sonochemically processed cationic nanocapsules: efficient antimicrobials with membrane disturbing capacity. Biomacromolecules 15:1365–1374. doi:10.1021/bm4018947
Follmann HD, Martins AF, Gerola AP, Burgo TAL, Nakamura CV, Rubira AF, Muniz EC (2012) Antiadhesive and antibacterial multilayer films via layer-by-layer assembly of TMC/heparin complexes. Biomacromolecules 13:3711–3722. doi:10.1021/bm3011962
Fu J, Ji J, Yuan W, Shen J (2005) Construction of anti-adhesive and antibacterial multilayer films via layer-by-layer assembly of heparin and chitosan. Biomaterials 26:6684–6692. doi:10.1016/j.biomaterials.2005.04.034
Gilles S (2007) Chemical modification of silicon surfaces for the application in soft lithography. Dissertation, University of Freiburg
Hammond PT (2004) Form and function in multilayer assembly: new applications at the nanoscale. Adv Mater 16:1271–1293. doi:10.1002/adma.200400760
Harmsen M, Yang L, Pamp SJ, Tolker-Nielsen T (2010) An update on Pseudomonas aeruginosa biofilm formation, tolerance, and dispersal. FEMS Immunol Med Microbiol 59:253–268. doi:10.1111/j.1574-695X.2010.00690.x
Hong S, Leroueil PR, Janus EK, Peters JL, Kober MM, Islam MT, Orr BG, Baker JR, Banaszak HMM (2006) Interaction of polycationic polymers with supported lipid bilayers and cells: nanoscale hole formation and enhanced membrane permeability. Bioconjugate Chem 17:728–734. doi:10.1021/bc060077y
Hong KW, Koh CL, Sam CK, Yin WF, Chan KG (2012) Quorum quenching revisited—from signal decays to signalling confusion. Sensors 12:4661–4696. doi:10.3390/s120404661
Horcas I, Fernández R, Gómez-Rodríguez JM, Colchero J, Gómez-Herrero J, Baro AM (2007) WSXM: A software for scanning probe microscopy and a tool for nanotechnology. Rev Sci Instrum 78:1–7. doi:10.1063/1.2432410
Ivanova K, Fernandes MM, Tzanov T (2013) Current advances on bacterial pathogenesis inhibition and treatment strategies. In: Mendez-Vilas A (ed) Microbial pathogens and strategies for combating them: science, technology and education. Formatex Research Center, Badajoz, pp 322–336
Jacobsen SM, Stickler DJ, Mobley HLT, Shirtliff ME (2008) Complicated catheter-associated urinary tract infections due to Escherichia coli and Proteus mirabilis. Clin Microbiol Rev 21:26–59. doi:10.1128/CMR. 00019-07
Kalia VC (2013) Quorum sensing inhibitors: an overview. Biotechnol Adv 31:224–245. doi:10.1016/j.biotechadv.2012.10.004
Kong J, Yu S (2007) Fourier transform infrared spectroscopic analysis of protein secondary structures. Acta Biochim Biophys Sin 39:549–559. doi:10.1111/j.1745-7270.2007.00320.x
Kong W, Zhang X, Gao ML, Zhou H, Li W, Shen JC (1994) A new kind of immobilized enzyme multilayer based on cationic and anionic interaction. Macromol Rapid Commun 15:405–409. doi:10.1002/marc.1994.030150503
Lawrence EL, Turner IG (2005) Materials for urinary catheters: a review of their history and development in the UK. J Biomed Eng 27:443–453. doi:10.1016/j.medengphy.2004.12.013
Lee SJ, Kim SW, Cho YH, Shin WS, Lee SE, Kim CS, Hong SJ, Chung BH, Kim JJ, Yoon MS (2004) A comparative multicentre study on the incidence of catheter-associated urinary tract infection between nitrofurazone-coated and silicone catheters. Int J Antimicrob Agents 24(SUPPL 1):65–69. doi:10.1016/j.ijantimicag.2004.02.013
Lellouche J, Friedman A, Lahmi R, Gedanken A, Banin E (2012) Antibiofilm surface functionalization of catheters by magnesium fluoride nanoparticles. Int J Nanomed 7:1175–1188. doi:10.2147/IJN.S26770
Li X, Li P, Saravanan R, Basu A, Mishra B, Lim SH, Su X, Tambyah PA, Leong SSJ (2014) Antimicrobial functionalization of silicone surfaces with engineered short peptides having broad spectrum antimicrobial and salt-resistant properties. Acta Biomater 10:258–266. doi:10.1016/j.actbio.2013.09.009
Lin YH, Xu JL, Hu J, Wang LH, Ong SL, Renton Leadbetter JR, Zhang LH (2003) Acyl-homoserine lactone acylase from Ralstonia strain XJ12B represents a novel and potent class of quorum-quenching enzymes. Mol Microbiol 47:849–860. doi:10.1046/j.1365-2958.2003.03351.x
Lopez-Perez PM, da Silva RMP, Serra C, Pashkuleva I, Reis RL (2010) Surface phosphorylation of chitosan significantly improves osteoblast cell viability, attachment and proliferation. J Mater Chem 20:483–491. doi:10.1039/b911854c
Onda M, Ariga K, Kunitake T (1999) Activity and stability of glucose oxidase in molecular films assembled alternately with polyions. J Biosci Bioeng 87:69–75. doi:10.1016/S1389-1723(99)80010-3
Palmer J, Flint S, Brooks J (2007) Bacterial cell attachment, the beginning of a biofilm. J Ind Microbiol Biotechnol 34:577–588. doi:10.1007/s10295-007-0234-4
Pavlukhina SV, Kaplan JB, Xu L, Chang W, Yu X, Madhyastha S, Yakandawala N, Mentbayeva A, Khan B, Sukhishvili SA (2012) Noneluting enzymatic antibiofilm coatings. ACS Appl Mater Interface 4:4708–4716. doi:10.1021/am3010847
Rajakaruna G, Harber M (2014) Urinary tract infection. In: Harber M (ed) Practical nephrology. Springer London, London, pp 395–411
Rasko DA, Sperandio V (2010) Anti-virulence strategies to combat bacteria-mediated disease. Nat Rev Drug Discov 9:117–128. doi:10.1038/nrd3013
Ravn L, Christensen AB, Molin S, Givskov M, Gram L (2001) Methods for detecting acylated homoserine lactones produced by Gram-negative bacteria and their application in studies of AHL-production kinetics. J Microbiol Methods 44:239–251. doi:10.1016/S0167-7012(01)00217-2
Richards JJ, Melander C (2009) Controlling bacterial biofilms. ChemBioChem 10:2287–2294. doi:10.1002/cbic.200900317
Richert L, Lavalle P, Payan E, Shu XZ, Prestwich GD, Stoltz JF, Schaaf P, Voegel JC, Picart C (2003) Layer by Layer buildup of polysaccharide films: physical chemistry and cellular adhesion aspects. Langmuir 20:448–458. doi:10.1021/la035415n
Sakr OS, Borchard G (2013) Encapsulation of enzymes in Layer-by-Layer (LbL) structures: latest advances and applications. Biomacromolecules 14:2117–2135. doi:10.1021/bm400198p
Sakuragi Y, Kolter R (2007) Quorum-sensing regulation of the biofilm matrix genes (pel) of Pseudomonas aeruginosa. J Bacteriol 189:5383–5386. doi:10.1128/jb.00137-07
Sarkar K, Kundu PP (2012) Preparation of low molecular weight N-maleated chitosan-graft-PAMAM copolymer for enhanced DNA complexation. Int J Biol Macromol 51:859–867. doi:10.1016/j.ijbiomac.2012.06.038
Sio CF, Otten LG, Cool RH, Diggle SP, Braun PG, Bos R, Daykin M, Cámara M, Williams P, Quax WJ (2006) Quorum quenching by an N-acyl-homoserine lactone acylase from Pseudomonas aeruginosa PAO1. Infect Immun 74:1673–1682. doi:10.1128/iai. 74.3.1673-1682.2006
Steindler L, Venturi V (2007) Detection of quorum-sensing N-acyl homoserine lactone signal molecules by bacterial biosensors. FEMS Microbiol Lett 266:1–9. doi:10.1111/j.1574-6968.2006.00501.x
Stickler DJ, Morris NS, Winters C (1999) Simple physical model to study formation and physiology of biofilms on urethral catheters. In: Ron JD (ed) Methods in enzymology. Elsevier, London, pp 494–501
Trautner BW (2010) Management of catheter-associated urinary tract infection. Curr Opin Infect Dis 23:76–82. doi:10.1097/QCO.0b013e328334dda8
UNE-EN 1616 (1997) Sterile Urethral Catheters for Single Use
Webb JS, Thompson LS, James S, Charlton T, Tolker-Nielsen T, Koch B, Givskov M, Kjelleberg S (2003) Cell death in Pseudomonas aeruginosa biofilm development. J Bacteriol 185:4585–4592. doi:10.1128/JB.185.15.4585-4592.2003
Xu F, Byun T, Dussen HJ, Duke KR (2003) Degradation of N-acylhomoserine lactones, the bacterial quorum-sensing molecules, by acylase. J Biotechnol 101:89–96. doi:10.1016/S0168-1656(02)00305-X
Youshko MI, Van Langen LM, Sheldon RA, Švedas VK (2004) Application of aminoacylase I to the enantioselective resolution of α-amino acid esters and amides. Tetrahedron Asymmetry 15:1933–1936. doi:10.1016/j.tetasy.2004.05.018
Acknowledgments
This research was performed in the framework of the European project “Novel approaches for prevention of biofilms formed on medical indwelling devices, e.g. catheters” (FP7-27840). K.I. thanks Generalitat de Catalunya for providing her PhD grant (FI_B1 00188), and M.M.F. acknowledges the Marie-Curie Intra-European Fellowship (IEF) NanoQuench (FP7-331416).
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Ivanova, K., Fernandes, M.M., Mendoza, E. et al. Enzyme multilayer coatings inhibit Pseudomonas aeruginosa biofilm formation on urinary catheters. Appl Microbiol Biotechnol 99, 4373–4385 (2015). https://doi.org/10.1007/s00253-015-6378-7
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DOI: https://doi.org/10.1007/s00253-015-6378-7