Difference in formation of a dental multi-species biofilm according to substratum direction.

OBJECTIVES: The aim of this study was to investigate the difference in dental biofilm formation according to substratum direction, using an artificial biofilm model. METHODS: A three-species biofilm, consisting of Streptococcus mutans, Streptococcus oralis, and Actinomyces naeslundii, was formed on saliva-coated hydroxyapatite (sHA) discs oriented in three directions: downward (the discs placed in the direction of gravity), vertical (the discs placed parallel to the direction of gravity), and upward (the discs placed in opposite direction of gravity). The biofilms at 22 h and 46 h of age were analyzed using microbiological and biochemical methods, fluorescence-based assays, and scanning electron microscopy to investigate difference in bacterial adhesion, early and mature biofilm formation. RESULTS: The biofilms formed in the upward direction displayed the most complex structure, with the highest number and biovolume of bacteria, as well as the lowest pH conditions at both time points. The vertical and downward directions, however, had only scattered and small bacterial colonies. In the 22-h-old biofilms, the proportion of S. oralis was similar to, or slightly higher than, that of S. mutans in all directions of substratum surfaces. However, in the 46-h-old biofilms, S. mutans became the dominant bacteria in all directions, especially in the vertical and upward directions. CONCLUSIONS: The direction of the substratum surface could impact the proportion of bacteria and cariogenic properties of the multi-species biofilm. Biofilms in an upward direction may exhibit a higher cariogenic potential, followed by those in the vertical and downward directions, which could be related to gravity.

The reciprocal interaction between fluoride release of glass ionomers and acid production of Streptococcus mutans biofilm.

Objectives: The aim of this study was to demonstrate the mode of action of glass ionomers (G-Is) against cariogenic biofilms in the slow fluoride release phase by analyzing the reciprocal interaction between fluoride release from G-Is and acid production of Streptococcus mutans biofilm. Methods: G-Is discs in the slow fluoride release phase were prepared and 51 h-old S. mutans biofilms were formed on these discs. The interrelationship between the acid production of the biofilm and the fluoride release of the G-Is discs was investigated by analyzing both factors simultaneously during the biofilm formation period. The composition of the 51 h-old biofilms was then examined using microbiological, biochemical, and confocal laser scanning microscopic methods. Results: Acid production by the cariogenic biofilm, particularly at < pH 5, promotes G-Is fluoride release. Conversely, G-Is fluoride release inhibits the acid production of the cariogenic biofilm. This reciprocal interaction results in the reduction of virulence such as extracellular polysaccharides formation and cariogenic biofilm bio-mass, which may reduce the potential of secondary caries development around G-Is. Conclusions: These results suggest that G-Is may play a role in preventing the development of secondary caries during the slow fluoride release phase.

Relationship between sucrose concentration and bacteria proportion in a multispecies biofilm: Short title: Sucrose challenges to a multispecies biofilm.

Objective: The aim of this study was to evaluate the relationship between sucrose concentration and bacteria proportion in a multispecies biofilm model. Methods: Streptococcus mutans (S. mutans), Streptococcus oralis (S. oralis), and Actinomyces naeslundii (A. naeslundii) were chose to form a multispecies biofilm. Different concentration (0-40%) of sucrose was introduced to the multispecies biofilm 3 times per day (30 min per time). And then the bacteria proportion and acid production of the biofilms were analyzed. Results: Increasing sucrose level increased CFU count of S. mutans up to a certain concentration (5% sucrose), after which the number of S. mutans slightly decreased, but the CFU counts of S. oralis and A. naeslundii continually decreased with sucrose concentration increase, especially, from 5% sucrose, the reduction was significant, and S. mutans became the dominant species in the biofilms. Furthermore, the acid production ability of the multispecies biofilm gradually increased and slightly decreased with sucrose concentration increased, and the turning concentration was 5%. Conclusion: Our findings suggest that increasing sucrose level could increase the competitiveness of S. mutans in the multispecies biofilm, which may shift the biofilm to a more cariogenic one, and 5% sucrose formed a most cariogenic biofilm in this study.

Sucrose challenges to Streptococcus mutans biofilms and the curve fitting for the biofilm changes.

The relationship between sugar level and development of dental caries has long been a main topic in dentistry. However, as a ubiquitous component of the modern diet, sucrose is mainly derived from three meals a day, rather than a long time exposure. In this study, various concentrations of sucrose were provided to Streptococcus mutans biofilms for 1 h per exposure (three times per day) to imitate a human meal pattern. And then the relationship between sucrose concentration and changes in the treated biofilms was determined. The results indicated that the components and acid production of the treated biofilms changed in a second-order polynomial curve pattern with sucrose concentration increase, which were confirmed by CLSM and SEM analyses. However, gene expression related to extracellular polysaccharides (EPS) formation, acid production and tolerance was up-regulated with sucrose concentration increase, which might have been due to compensation for the decrease in EPS formation and acid production by the biofilms at higher concentrations of sucrose. These findings suggest that sucrose in the range of 1%-5% can support the highest acid production and accumulation of S. mutans biofilms, which may further increase its cariogenic potential. However, additional studies are required to confirm the relationships in human cariogenic biofilms.

Difference in virulence and composition of a cariogenic biofilm according to substratum direction.

The aim of this study was to investigate the difference in composition and virulence of Streptococcus mutans biofilms according to substratum direction. S. mutans biofilms (46-h-old) were formed on three different saliva-coated hydroxyapatite (sHA) disc direction groups: downward (discs placed in the direction of gravity), vertical (discs placed parallel to gravity direction), and upward (discs placed opposite to gravity). The 46-h-old biofilms on sHA discs in the upward direction showed the highest biofilm accumulation, colony forming unit (CFU) count, and extracellular polysaccharide (EPS) amount, followed by those in the vertical and downward directions. In the confocal laser scanning microscopy (CLSM) study, the biofilms in the upward direction also showed the highest bacterial count (live or dead cells) and EPS biovolume. Scanning electron microscopy (SEM) analysis confirmed the microbiological and biochemical results. In addition, biofilm density and acid production were higher in the upward direction than those in the other directions. Our findings suggest that substratum direction, which might be related to gravity, strongly influences the formation and virulence of cariogenic biofilms and subsequent initiation of dental caries. Collectively, the differences in the formation and virulence of cariogenic biofilms are related to the direction of tooth surface (occlusal surfaces of mandibular teeth > proximal surfaces > occlusal surfaces of maxillary teeth).

Trend Analysis of Studies on Water Fluoridation Related to Dental Caries in PubMed.

Water fluoridation has been cited as one of the top 10 public health achievements of the 20th century. Herein, we analyzed water fluoridation articles related to dental caries published in PubMed between 1950 and 2016 using informetrics and linguistic methods to investigate trends in the studies. To this aim, queries such as "dental caries and (water fluoridation or fluoridated water)," "dental caries and (fluoride or fluoridation)," and "dental caries" were submitted to PubMed to retrieve information about articles on water fluoridation within the area of dental caries and fluoride - their titles, abstracts, publication dates, author affiliations, and publication journals. This article information was then collected by an automatic web crawler and examined through informetrics and linguistic analyses. It was found that the number of articles concerned with water fluoridation and dental caries was 3,381 and declined over time after 1970. The articles were published by 750 journals - most notably, Community Dentistry and Oral Epidemiology and Caries Research. With regard to the geographic distribution of the authors, Europe and North America, especially the USA and UK, accounted for 59.9% of the articles published during the years 1987 to 2015, though there was a sharp increase in the number of authors in Oceania and Asia in recent years. In the titles and abstracts of the articles, "community" and "fluorosis" were mentioned more frequently than the other key terms selected in this study, regardless of the period examined. Our findings may allow one to assess how the research on water fluoridation has evolved over the past several decades.

Effect of brief periodic fluoride treatments on the virulence and composition of a cariogenic biofilm.

The present study investigated the effect of periodic 1-min fluoride treatments on Streptococcus mutans biofilms and then determined the relationship between anti-biofilm activity, treatment frequency, and fluoride concentration using a linear-fitting procedure. S. mutans biofilms were periodically treated (1-min/treatment) with fluoride during biofilm formation and analyzed using microbiological methods, confocal microscopy, and real-time PCR. The results indicated that reductions in the dry weight and acidogenicity of biofilms due to periodic fluoride treatment occurred in a concentration dependent manner. The reduction in dry weight without affecting bacterial cell viability was observed mainly due to the inhibitory effect of fluoride on gtfB and gtfC gene expression, which suppresses EPS production and avoids reduction of the pH below the critical point on the tooth surface. This study suggests that brief periodic exposure to appropriate fluoride concentrations through mouthwashes and toothpastes may affect the virulence and composition of cariogenic biofilms and subsequently prevent dental caries.