Advertisement

Decreased Bacterial Adherence and Biofilm Growth on Surfaces Coated with a Solution of Benzalkonium Chloride

Published:April 27, 2012DOI:https://doi.org/10.1016/j.joen.2012.03.012

      Abstract

      Introduction

      Secondary biofilm formation by oral bacteria after breakdown/fracture of temporary or permanent restorations imposes a challenge to the outcome of root canal treatment. This study focuses on benzalkonium chloride (BAK) coating on dentin or polystyrene surfaces and its influence on the early adhesion and biofilm formation by oral and root canal bacteria.

      Methods

      Microbial adhesion and biofilm growth on surfaces coated with BAK were analyzed qualitatively with a dentin disk model and quantitatively with a mini-flow cell biofilm model. Cell viability and total biovolume were analyzed by the LIVE/DEAD technique. The repelling effect of surfaces coated with BAK was compared with NaOCl. Uncoated surfaces were used as controls.

      Results

      Scanning electron microscope images in the dentin disk model revealed that very sparse biofilms were formed on NaOCl- and BAK-coated dentin surfaces. In contrast, biofilms formed on uncoated dentin were clearly visible as numerous irregularly distributed aggregates of rods and cocci. In the mini-flow cell system, confocal laser scanning microscope analysis confirmed that biofilms formed on NaOCl- and BAK-coated surfaces showed significantly less adhesion (2 hours) and biovolume accumulation (24 and 96 hours) compared with the uncoated controls (P < .01). Furthermore, cell viability assessments showed that on uncoated controls the viability measurements were high (>89%) as well as on BAK-coated surfaces (88% viable cells). However, cell viability was significantly reduced on NaOCl-coated surfaces (59% viable cells).

      Conclusions

      This study illustrates that surface coating with a surfactant solution containing BAK does not cause cell membrane damage but might interfere with cell mechanisms of adhesion. Investigations into the clinical utility of BAK as an antibiofilm medication are warranted.

      Key Words

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Journal of Endodontics
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Siqueira Jr., J.F.
        Microbial causes of endodontic flare-ups.
        Int Endod J. 2003; 36: 453-463
        • Siqueira Jr., J.F.
        • Rocas I.N.
        • Favieri A.
        • Abad E.C.
        • Castro A.J.
        • Gahyva S.M.
        Bacterial leakage in coronally unsealed root canals obturated with 3 different techniques.
        Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2000; 90: 647-650
        • Chavez de Paz L.E.
        Redefining the persistent infection in root canals: possible role of biofilm communities.
        J Endod. 2007; 33 (Erratum in: J Endod 2007;33(11):1289): 652-662
        • Lau P.C.
        • Dutcher J.R.
        • Beveridge T.J.
        • Lam J.S.
        Absolute quantitation of bacterial biofilm adhesion and viscoelasticity by microbead force spectroscopy.
        Biophys J. 2009; 96: 2935-2948
        • Herzberg M.
        • Elimelech M.
        Physiology and genetic traits of reverse osmosis membrane biofilms: a case study with Pseudomonas aeruginosa.
        ISME J. 2008; 2: 180-194
        • Gomez-Suarez C.
        • Busscher H.J.
        • van der Mei H.C.
        Analysis of bacterial detachment from substratum surfaces by the passage of air-liquid interfaces.
        Appl Environ Microbiol. 2001; 67: 2531-2537
        • Murga R.
        • Miller J.M.
        • Donlan R.M.
        Biofilm formation by gram-negative bacteria on central venous catheter connectors: effect of conditioning films in a laboratory model.
        J Clin Microbiol. 2001; 39: 2294-2297
        • Park Y.
        • Park S.N.
        • Park S.C.
        • et al.
        Antibiotic activity and synergistic effect of antimicrobial peptide against pathogens from a patient with gallstones.
        Biochem Biophys Res Commun. 2004; 321: 631-637
        • Gottenbos B.
        • Grijpma D.W.
        • van der Mei H.C.
        • Feijen J.
        • Busscher H.J.
        Antimicrobial effects of positively charged surfaces on adhering Gram-positive and Gram-negative bacteria.
        J Antimicrob Chemother. 2001; 48: 7-13
        • Tsibouklis J.
        • Stone M.
        • Thorpe A.A.
        • et al.
        Preventing bacterial adhesion onto surfaces: the low-surface-energy approach.
        Biomaterials. 1999; 20: 1229-1235
        • Cloete T.E.
        • Oosthuizen D.J.
        The role of extracellular exopolymers in the removal of phosphorus from activated sludge.
        Water Res. 2001; 35: 3595-3598
        • Meylheuc T.
        • Methivier C.
        • Renault M.
        • Herry J.M.
        • Pradier C.M.
        • Bellon-Fontaine M.N.
        Adsorption on stainless steel surfaces of biosurfactants produced by gram-negative and gram-positive bacteria: consequence on the bioadhesive behavior of Listeria monocytogenes.
        Colloids Surf B Biointerfaces. 2006; 52: 128-137
        • Pereira H.A.
        Novel therapies based on cationic antimicrobial peptides.
        Curr Pharm Biotechnol. 2006; 7: 229-234
        • Splendiani A.
        • Livingston A.G.
        • Nicolella C.
        Control of membrane-attached biofilms using surfactants.
        Biotechnol Bioeng. 2006; 94: 15-23
        • Pozarowska D.
        • Pozarowski P.
        Benzalkonium chloride (BAK) induces apoptosis or necrosis, but has no major influence on the cell cycle of Jurkat cells.
        Folia Histochem Cytobiol. 2011; 49: 225-230
        • Nomura Y.
        • Bhawal U.K.
        • Nishikiori R.
        • Sawajiri M.
        • Maeda T.
        • Okazaki M.
        Effects of high-dose major components in oral disinfectants on the cell cycle and apoptosis in primary human gingival fibroblasts in vitro.
        Dent Mater J. 2010; 29: 75-83
        • Haapasalo M.
        • Ørstavik D.
        In vitro infection and disinfection of dentinal tubules.
        J Dent Res. 1987; 66: 1375-1379
        • Schaudinn C.
        • Carr G.
        • Gorur A.
        • Jaramillo D.
        • Costerton J.W.
        • Webster P.
        Imaging of endodontic biofilms by combined microscopy (FISH/cLSM - SEM).
        J Microsc. 2009; 235: 124-127
        • Chávez de Paz L.E.
        • Dahlén G.
        • Molander A.
        • Möller A.
        • Bergenholtz G.
        Bacteria recovered from teeth with apical periodontitis after antimicrobial endodontic treatment.
        Int Endod J. 2003; 36: 500-508
        • Chavez de Paz L.E.
        • Bergenholtz G.
        • Svensäter G.
        The effects of antimicrobials on endodontic biofilm bacteria.
        J Endod. 2010; 36: 70-77
        • Chavez de Paz L.E.
        • Hamilton I.R.
        • Svensäter G.
        Oral bacteria in biofilms exhibit slow reactivation from nutrient deprivation.
        Microbiology. 2008; 154: 1927-1938
        • Chavez de Paz L.E.
        Image analysis software based on color segmentation for characterization of viability and physiological activity of biofilms.
        Appl Environ Microbiol. 2009; 75: 1734-1739
        • Pieper D.H.
        • Reineke W.
        Engineering bacteria for bioremediation.
        Curr Opin Biotechnol. 2000; 11: 262-270
        • Gilbert P.
        • Moore L.E.
        Cationic antiseptics: diversity of action under a common epithet.
        J Appl Microbiol. 2005; 99: 703-715
        • Shen Y.
        • Stojicic S.
        • Qian W.
        • Olsen I.
        • Haapasalo M.
        The synergistic antimicrobial effect by mechanical agitation and two chlorhexidine preparations on biofilm bacteria.
        J Endod. 2010; 36: 100-104
        • Shen Y.
        • Stojicic S.
        • Haapasalo M.
        Bacterial viability in starved and revitalized biofilms: comparison of viability staining and direct culture.
        J Endod. 2010; 36: 1820-1823
        • Shen Y.
        • Stojicic S.
        • Haapasalo M.
        Antimicrobial efficacy of chlorhexidine against bacteria in biofilms at different stages of development.
        J Endod. 2011; 37: 657-661
        • George S.
        • Kishen A.
        • Song K.P.
        The role of environmental changes on monospecies biofilm formation on root canal wall by Enterococcus faecalis.
        J Endod. 2005; 31: 867-872
        • Kishen A.
        • Upadya M.
        • Tegos G.P.
        • Hamblin M.R.
        Efflux pump inhibitor potentiates antimicrobial photodynamic inactivation of Enterococcus faecalis biofilm.
        Photochem Photobiol. 2010; 86: 1343-1349
        • Darouiche R.O.
        Prevention of vascular catheter-related infections.
        Neth J Med. 1999; 55: 92-99
        • Darouiche R.O.
        • Raad I.I.
        • Heard S.O.
        • et al.
        A comparison of two antimicrobial-impregnated central venous catheters: Catheter Study Group.
        N Engl J Med. 1999; 340: 1-8
        • Edmiston Jr., C.E.
        • Goheen M.P.
        • Seabrook G.R.
        • et al.
        Impact of selective antimicrobial agents on staphylococcal adherence to biomedical devices.
        Am J Surg. 2006; 192: 344-354