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Chlorhexidine rinsing inhibits biofilm formation and causes biofilm disruption on dental enamel in situ

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Abstract

Objectives

This in situ study aims to evaluate the effects of chlorhexidine (CHX) mouth rinsing on biofilm formation and moreover on the disruption of existing mature dental biofilms.

Methods

Biofilms were formed in situ by five volunteers on bovine enamel specimens fixed to individual acrylic splints. For biofilm formation analysis, the volunteers intraorally exposed the splint for 48 h. Mouth rinsing using 10 ml of 0.2% CHX or water as control was performed for 30 s every 12 h. For analysis of biofilm disruption, the biofilm was formed on enamel specimens for 48 h. Then, the first CHX rinse was carried out. A second rinse followed after an additional 12 h, again for 30 s using 10 ml of 0.2% CHX. Biofilm vitality was imaged by fluorescence microscopy after vital fluorescence staining. Additionally, the ultrastructure of the biofilm was examined by transmission electron microscopy.

Results

Rinses with 0.2% CHX significantly reduced biofilm formation on enamel. Both biofilm colonization and vitality were dramatically impaired. Moreover, a considerable biofilm disruption induced by the CHX rinses was observed. Remarkably, a single application of CHX to a 48-h mature biofilm causes biofilm ultrastructure alterations and induces a substantial reduction in biofilm thickness and bacterial vitality.

Conclusions

CHX mouth rinses induced a significant inhibition of biofilm formation on native enamel. Furthermore, an important biofilm disrupting effect under in situ conditions was detected.

Clinical Relevance: CHX rinses could be used as a short-term treatment protocol for biofilm management focused on patients unable to reach adequate oral hygiene.

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Acknowledgements

The authors thank Birgit Leis and Belinda König for technical assistance with the samples processing for TEM analysis.

Funding

This study has been funded by the German Research Foundation (DFG, SFB 1027).

Author information

Author notes

  1. Miryam Martínez-Hernández and Bashar Reda contributed equally to this work.

Authors and Affiliations

  1. Clinic of Operative Dentistry, Periodontology and Preventive Dentistry, University Hospital, Saarland University, Building 73, 66421, Homburg/Saar, Germany

    Miryam Martínez-Hernández, Bashar Reda & Matthias Hannig

  2. Facultad de Odontología, División de Estudios de Posgrado e Investigación, Universidad Nacional Autónoma de México, Circuito exterior s/n, Ciudad Universitaria, 04510, CDMX, México

    Miryam Martínez-Hernández

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  1. Miryam Martínez-Hernández
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  2. Bashar Reda
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Correspondence to Matthias Hannig.

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Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were approved by the Ethical Committee of Saarland Medical Association, Germany (238/03, 2016). The study was also in accordance with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

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Informed consent was obtained from all individual participants included in the study.

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Martínez-Hernández, M., Reda, B. & Hannig, M. Chlorhexidine rinsing inhibits biofilm formation and causes biofilm disruption on dental enamel in situ. Clin Oral Invest 24, 3843–3853 (2020). https://doi.org/10.1007/s00784-020-03250-3

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  • DOI: https://doi.org/10.1007/s00784-020-03250-3

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