Development of In Vitro Denture Biofilm Models for Halitosis Related Bacteria and their Application in Testing the Efficacy of Antimicrobial Agents

Tingxi Wu 1, Xuesong He 1, Hongyang Lu 2, David J Bradshaw 3, Alyson Axe 3, Zvi Loewy 4, Honghu Liu 1, Wenyuan Shi 1, Renate Lux 1, *
1 School of Dentistry, University of California Los Angeles, Los Angeles, USA
2 West China School of Stomatology, Sichuan University, Chengdu, China
3 GlaxoSmithKline, Family Oral Health, Weybridge, UK
4 Department of Pharmaceutical and Biomedical Sciences, Touro College of Pharmacy, New York, NY, USA

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© Wu et al.; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

* Address correspondence to this author at the Section of Oral Biology in the Division of Oral Biology and Medicine, School of Dentistry, University of California Los Angeles, 33-080 CHS, 10833 Le Conte Avenue, Los Angeles, CA, USA, 90095-1668: Tel: +13102065660; Fax: +13107947109; E-mail:


Objective : Since dentures can serve as a reservoir for halitosis-causing oral bacteria, halitosis development is a concern for denture wearers. In this study, we surveyed the prevalence of four selected halitosis-related species (Fusobacterium nucleatum, Tannerella forsythia, Veillonella atypica and Klebsiella pneumoniae) in clinical denture plaque samples, and developed denture biofilm models for these species in vitro to facilitate assessment of antimicrobial treatment efficacy. Design : Denture plaque from ten healthy and ten denture stomatitis patients was screened for the presence of aforementioned four species by PCR. Biofilm formation by these halitosis-associated species on the surfaces of denture base resin (DBR) discs was evaluated by crystal violet staining and confocal laser scanning microscopy. The efficacy of denture cleanser treatment on these mono-species biofilms was evaluated by colony counting. Results : 80% of the subjects in the denture stomatitis group and 60% in the healthy group contained at least one of the targeted halitosis-related species in their denture plaque. All halitosis species tested were able to form biofilms on DBR disc surfaces to varying degrees. These in vitro mono-species resin biofilm models were used to evaluate the efficacy of denture cleansers, which exhibited differential efficacies. When forming biofilms on resin surfaces, the halitosis-related species displayed enhanced resistance to denture cleansers compared with their planktonic counterparts. Conclusion : The four selected halitosis-related bacterial species examined in this study are present on the majority of dentures. The mono-species biofilm models established on DBR discs for these species are an efficient screening tool for dental product evaluation.

Keywords: Antimicrobial treatment, biofilms, denture, halitosis, model.