Casein Phosphopeptide-Amorphous Calcium Phosphate Reduces Streptococcus mutans Biofilm Development on Glass Ionomer Cement and Disrupts Established Biofilms.

Stuart G Dashper,Joseph E A Palamara,Helen Myroforidis,Eric C Reynolds,Deanne V Catmull,Ilya Zalizniak,N Laila Huq,Sze-Wei Liu,Abdelwahab Omri

doi:10.1371/journal.pone.0162322

Stuart G Dashper, Joseph E A Palamara + Show 7 more

Open Access

https://doi.org/10.1371/journal.pone.0162322

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Journal: PloS one	Publication Date: Sep 2, 2016
Citations: 30	License type: CC BY 4.0

Affiliation: University of Melbourne

Abstract

Glass ionomer cements (GIC) are dental restorative materials that are suitable for modification to help prevent dental plaque (biofilm) formation. The aim of this study was to determine the effects of incorporating casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) into a GIC on the colonisation and establishment of Streptococcus mutans biofilms and the effects of aqueous CPP-ACP on established S mutans biofilms. S. mutans biofilms were either established in flow cells before a single ten min exposure to 1% w/v CPP-ACP treatment or cultured in static wells or flow cells with either GIC or GIC containing 3% w/w CPP-ACP as the substratum. The biofilms were then visualised using confocal laser scanning microscopy after BacLight LIVE/DEAD staining. A significant decrease in biovolume and average thickness of S. mutans biofilms was observed in both static and flow cell assays when 3% CPP-ACP was incorporated into the GIC substratum. A single ten min treatment with aqueous 1% CPP-ACP resulted in a 58% decrease in biofilm biomass and thickness of established S. mutans biofilms grown in a flow cell. The treatment also significantly altered the structure of these biofilms compared with controls. The incorporation of 3% CPP-ACP into GIC significantly reduced S. mutans biofilm development indicating another potential anticariogenic mechanism of this material. Additionally aqueous CPP-ACP disrupted established S. mutans biofilms. The use of CPP-ACP containing GIC combined with regular CPP-ACP treatment may lower S. mutans challenge.

Highlights

Dental caries is one of the most prevalent chronic diseases of humans and affects the majority of individuals globally [1]
Fuji VII and Fuji VII Enhanced Protection (EP) Glass ionomer cements (GIC) blocks were placed into tissue culture wells containing 25% ASM and inoculated with S. mutans
The inclusion of 3% Casein phosphopeptide-stabilised amorphous calcium phosphate (CPP-ACP) in Fuji VII EP significantly reduced the biovolume of the S. mutans biofilm by 50% relative to the Fuji VII control and average thickness of the biofilm was reduced by 66% (Table 1 and Fig 2)

Summary

Introduction

Dental caries is one of the most prevalent chronic diseases of humans and affects the majority of individuals globally [1]. The disease is a dynamic process that is initiated in the bacterial biofilm (dental plaque) on the tooth surface by the production of organic acids from the fermentation of dietary sugar resulting in demineralisation of the tooth [2]. CPP-ACP Effects on S. mutans Biofilms requires a shift in plaque ecology to favour acidogenic and aciduric microbial species that is usually driven by frequent consumption of simple carbohydrates and is modified by host factors such as saliva [3]. Dental caries has a polymicrobial aetiology Streptococcus mutans is regarded as a major aetiological agent of this disease mainly due to its aciduric and highly acidogenic nature. In addition CPP-ACP incorporated into a sugar-free gum has been shown to slow the progression and enhance the regression of dental caries in a randomised, controlled clinical trial [17]

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