Abstract
Objectives
In this study, we aimed to develop a novel, sustained release varnish (SRV) for voice prostheses (VP) releasing chlorhexidine (CHX), for the prevention of biofilm formation caused by the common oral bacteria Streptococcus mutans on VP surfaces.
Methods
This study was performed in an in vitro model as a step towards future in vivo trials. VPs were coated with a SRV containing CHX (SRV-CHX) or SRV alone (placebo-SRV) that were daily exposed to S. mutans. The polymeric materials of SRV were composed of ethylcellulose and PEG-400. Biofilm formation was assessed by DNA quantification (qPCR), crystal violet staining, confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM), and kinetics experiments.
Results
The amount of DNA in the biofilms formed by S. mutans on VP surfaces coated once with SRV-CHX (1.024 ± 0.218 ng DNA/piece) was 58.5 ± 8.8% lower than that of placebo-SRV-coated VPs (2.465 ± 0.198 ng DNA/piece) after a 48-h exposure to S. mutans (p = 0.038). Reduced biofilm mass on SRV-CHX-coated VPs was visually confirmed by CLSM and SEM. CV staining of SRV-CHX single-coated VPs that have been exposed to S. mutans nine times showed a 98.1 ± 0.2% reduction in biofilm mass compared to placebo-SRV-coated VPs (p = 0.003). Kinetic experiments revealed that SRV-CHX triple-coated VPs could delay bacterial growth for 23 days.
Conclusions
Coating VPs with SRV-CHX has an inhibitory effect on biofilm formation and prevents bacterial growth in their vicinities. This study is a proof-of-principle that paves the way for developing new clinical means for reducing both VPs’ bacterial biofilm formation and device failure.
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Availability of data and material
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
Code availability
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References
Ajdić D, McShan WM, McLaughlin RE, Savić G, Chang J, Carson MB, Primeaux C, Tian R, Kenton S, Jia H, Lin S, Qian Y, Li S, Zhu H, Najar F, Lai H, White J, Roe BA, Ferretti JJ (2002) Genome sequence of Streptococcus mutans UA159, a cariogenic dental pathogen. Proc Natl Acad Sci U S A 99(22):14434–14439. https://doi.org/10.1073/pnas.172501299
Ameye D, Honraet K, Loose D, Vermeersch H, Nelis H, Remon JP (2005) Effect of a buccal bioadhesive nystatin tablet on the lifetime of a Provox silicone tracheoesophageal voice prosthesis. Acta Otolaryngol 125(3):304–306. https://doi.org/10.1080/00016480410022778
Bauters TG, Moerman M, Vermeersch H, Nelis HJ (2002) Colonization of voice prostheses by albicans and non-albicans Candida species. Laryngoscope 112(4):708–712
Bertl K, Zatorska B, Leonhard M, Rechenmacher-Strauss J, Roesner I, Schneider-Stickler B (2013) Oral microbial colonization in laryngectomized patients as a possible cofactor of biofilm formation on their voice prostheses. J Clin Periodontol 40(9):833–840. https://doi.org/10.1111/jcpe.12131
Bikiaris D, Koutris E, Karavas E (2007) New aspects in sustained drug release formulations. Recent Pat Drug Deliv Formul 1(3):201–213. https://doi.org/10.2174/187221107782331629
Block SS (2001) Disinfection, sterilization, and preservation. Lippincott Williams & Wilkins
Bueno MG, Urban VM, Barbério GS, da Silva WJ, Porto VC, Pinto L, Neppelenbroek KH (2015) Effect of antimicrobial agents incorporated into resilient denture relines on the Candida albicans biofilm. Oral Dis 21(1):57–65. https://doi.org/10.1111/odi.12207
Buijssen KJ, van der Laan BF, van der Mei HC, Atema-Smit J, van den Huijssen P, Busscher HJ, Harmsen HJ (2012) Composition and architecture of biofilms on used voice prostheses. Head Neck 34(6):863–871. https://doi.org/10.1002/hed.21833
Darouiche RO, Mansouri MD, Gawande PV, Madhyastha S (2008) Efficacy of combination of chlorhexidine and protamine sulphate against device-associated pathogens. J Antimicrob Chemother 61(3):651–657. https://doi.org/10.1093/jac/dkn006
Dijk F, Westerhof M, Busscher HJ, van Luyn MJ, van Der Mei HC (2000) In vitro formation of oropharyngeal biofilms on silicone rubber treated with a palladium/tin salt mixture. J Biomed Mater Res 51(3):408–412. https://doi.org/10.1002/1097-4636(20000905)51:3%3c408::aid-jbm15%3e3.0.co;2-m
Everaert EP, Mahieu HF, van de Belt-Gritter B, Peeters AJ, Verkerke GJ, van der Mei HC, Busscher HJ (1999) Biofilm formation in vivo on perfluoro-alkylsiloxane-modified voice prostheses. Arch Otolaryngol Head Neck Surg 125(12):1329–1332. https://doi.org/10.1001/archotol.125.12.1329
Everaert EP, van de Belt-Gritter B, van der Mei HC, Busscher HJ, Verkerke GJ, Dijk F, Mahieu HF, Reitsma A (1998) In vitro and in vivo microbial adhesion and growth on argon plasma-treated silicone rubber voice prostheses. J Mater Sci Mater Med 9(3):147–157. https://doi.org/10.1023/a:1008867619481
Friedman M, Golomb G (1982) New sustained release dosage form of chlorhexidine for dental use. I. Development and kinetics of release. J Periodontal Res 17(3):323–328. https://doi.org/10.1111/j.1600-0765.1982.tb01160.x
Gaonkar TA, Caraos L, Modak S (2007) Efficacy of a silicone urinary catheter impregnated with chlorhexidine and triclosan against colonization with Proteus mirabilis and other uropathogens. Infect Control Hosp Epidemiol 28(5):596–598. https://doi.org/10.1086/513449
Gibbons RJ (1989) Bacterial adhesion to oral tissues: a model for infectious diseases. J Dent Res 68(5):750–760. https://doi.org/10.1177/00220345890680050101
Hancock KL, Ward LNR, EC, (2013) Device life of the Provox Vega voice prosthesis. Eur Arch Otorhinolaryngol 270(4):1447–1453. https://doi.org/10.1007/s00405-012-2154-9
Kania RE, Lamers GE, van de Laar N, Dijkhuizen M, Lagendijk E, Tran Ba Huy P, Herman P, Hiemstra P, Grote JJ, Frijns J, Bloemberg GV (2010) Biofilms on tracheoesophageal voice prostheses: a confocal laser scanning microscopy demonstration of mixed bacterial and yeast biofilms. Biofouling 26(5):519–526
Koo H, Hayacibara MF, Schobel BD, Cury JA, Rosalen PL, Park YK, Vacca-Smith AM, Bowen WH (2003) Inhibition of Streptococcus mutans biofilm accumulation and polysaccharide production by apigenin and tt-farnesol. J Antimicrob Chemother 52(5):782–789. https://doi.org/10.1093/jac/dkg449
Kress P, Schäfer P, Schwerdtfeger FP (2006) Clinical use of a voice prosthesis with a flap valve containing silver oxide (Blom-Singer Advantage), biofilm formation, in-situ lifetime and indication. Laryngorhinootologie 85(12):893–896. https://doi.org/10.1055/s-2006-925292
Leonhard M, Schneider-Stickler B (2015) Voice prostheses, microbial colonization and biofilm formation. Adv Exp Med Biol 830:123–136. https://doi.org/10.1007/978-3-319-11038-7_8
Neu TR, De Boer CE, Verkerke GJ, Schutte HK, Rakhorst G, Van Der Meil HC, Busscher HJ (1994) Biofilm development in time on a silicone voice prosthesis - case study. Microb Ecol Health Dis 7(1):27–33
Periasamy S, Kolenbrander PE (2009) Mutualistic biofilm communities develop with Porphyromonas gingivalis and initial, early, and late colonizers of enamel. J Bacteriol 191(22):6804–6811. https://doi.org/10.1128/jb.01006-09
Radnai M, Whiley R, Friel T, Wright PS (2010) Effect of antifungal gels incorporated into a tissue conditioning material on the growth of Candida albicans. Gerodontology 27(4):292–296. https://doi.org/10.1111/j.1741-2358.2009.00337.x
Rodrigues L, van der Mei H, Banat IM, Teixeira J, Oliveira R (2006) Inhibition of microbial adhesion to silicone rubber treated with biosurfactant from Streptococcus thermophilus A. FEMS Immunol Med Microbiol 46(1):107–112. https://doi.org/10.1111/j.1574-695X.2005.00006.x
Segev G, Bankirer T, Steinberg D, Duvdevani M, Shapur NK, Friedman M, Lavy E (2013) Evaluation of urinary catheters coated with sustained-release varnish of chlorhexidine in mitigating biofilm formation on urinary catheters in dogs. J Vet Intern Med 27(1):39–46. https://doi.org/10.1111/j.1939-1676.2012.01027.x
Shapur NK, Duvdevani M, Friedman M, Zaks B, Gati I, Lavy E, Katz R, Landau EH, Pode D, Gofrit ON, Steinberg D (2012) Sustained release varnish containing chlorhexidine for prevention of biofilm formation on urinary catheter surface: in vitro study. J Endourol 26(1):26–31. https://doi.org/10.1089/end.2011.0140
Somogyi-Ganss E, Chambers MS, Lewin JS, Tarrand JJ, Hutcheson KA (2017) Biofilm on the tracheoesophageal voice prosthesis: considerations for oral decontamination. Eur Arch Otorhinolaryngol 274(1):405–413. https://doi.org/10.1007/s00405-016-4193-0
Steinberg D, Friedman M (2000) Development of sustained-release devices for modulation of dental plaque biofilm and treatment of oral infectious diseases. Drug Dev Res 50(3–4):555–565
Tićac B, Tićac R, Rukavina T, Kesovija PG, Pedisić D, Maljevac B, Starčević R (2010) Microbial colonization of tracheoesophageal voice prostheses (Provox2) following total laryngectomy. Eur Arch Otorhinolaryngol 267(10):1579–1586
Van Weissenbruch R, Albers FW, Bouckaert S, Nelis HJ, Criel G, Remon JP, Sulter AM (1997) Deterioration of the Provox™ silicone tracheoesophageal voice prosthesis: microbial aspects and structural changes. Acta Otolaryngol 117(3):452–458
Zelichenko G, Steinberg D, Lorber G, Friedman M, Zaks B, Lavy E, Hidas G, Landau EH, Gofrit ON, Pode D, Duvdevani M (2013) Prevention of initial biofilm formation on ureteral stents using a sustained releasing varnish containing chlorhexidine: in vitro study. J Endourol 27(3):333–337. https://doi.org/10.1089/end.2012.0193
Acknowledgements
The authors thank Dr. Mark Tarshish and Dr. Vitaly Gutkin (Hebrew University, Jerusalem, Israel) for the assistance with the CLSM and SEM, respectively. Menachem Gross, Fadi Ashqar, and Ronit Vogt Sionov have equal contribution to this study.
Funding
This study was supported by a grant from the Israeli Cancer Association (Ref Number—20180025).
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Contributions
Conceptualization: Menachem Gross, Michael Friedman, Doron Steinberg.
Data curation: Menachem Gross, Ron Eliashar, Ronit Vogt Sionov, Mark Feldman.
Formal analysis: Fadi Ashqar, Batya Zaks, Ronit Vogt Sionov, Mark Feldman.
Funding acquisition: Menachem Gross, Doron Steinberg.
Investigation: Menachem Gross, Fadi Ashqar, Ron Eliashar, Batya Zaks, Ronit Vogt Sionov, Mark Feldman, Doron Steinberg.
Methodology: Menachem Gross, Batya Zaks, Ronit Vogt Sionov, Mark Feldman Irith Gati, Doron Steinberg.
Project administration: Ron Eliashar, Doron Steinberg, Menachem Gross.
Resources: Michael Friedman, Doron Steinberg, Menachem Gross.
Software: Danielle Duanis-Assaf, Ronit Vogt Sionov.
Supervision: Michael Friedman, Doron Steinberg, Menachem Gross.
Validation: Danielle Duanis-Assaf, Ronit Vogt Sionov, Mark Feldman, Doron Steinberg.
Visualization: Mark Feldman, Danielle Duanis-Assaf, Ronit Vogt Sionov.
Writing—original draft: Menachem Gross.
Writing—review and editing: Menachem Gross, Doron Steinberg, Ronit Vogt Sionov.
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Highlights:
1. Speech rehabilitation with voice prosthesis is the treatment of choice in patients following total laryngectomy.
2. Biofilm development on voice prosthesis limits the lifetime of the product resulting in early replacement of the prosthesis.
3. Chlorhexidine has antiseptic and antifungal properties that prevent biofilm formation.
4. Sustained release varnish is a method for local slow release of an agent from a coated prosthesis or device, thereby prolonging the duration of the drug at the target site.
5. Utilization of sustained release varnish of chlorhexidine can prolong the lifetime of the VP via a delay of biofilm formation.
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Gross, M., Ashqar, F., Sionov, R.V. et al. Sustained release varnish containing chlorhexidine for prevention of Streptococcus mutans biofilm formation on voice prosthesis surface: an in vitro study. Int Microbiol 25, 177–187 (2022). https://doi.org/10.1007/s10123-021-00205-w
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DOI: https://doi.org/10.1007/s10123-021-00205-w