Effect of the Probiotic Lactobacillus rhamnosus LB21 on the Cariogenicity of Streptococcus mutans UA159 in a Dual-Species Biofilm Model

Despite promising results using probiotics, evidence of the preventive effect on enamel demineralization is insufficient and the cariogenic potential of probiotics is still controversial. Probiotics could affect biofilm formation and interfere with adherence, growth or coaggregation with Streptococcus mutans in biofilms. However, most of the studies have been conducted using planktonic bacteria. Hence, the aim of the study was to assess the effect of probiotic bacteria on the cariogenicity of S. mutans using an in vitro biofilm caries model on enamel. Single-species biofilms (S. mutans UA159, SM or Lactobacillus rhamnosus LB21, LB) or dual-species biofilms simultaneously inoculated (SM + LB) or LB inoculated 8 h after SM (SM → LB) were grown for 96 h. Biofilms were formed on bovine enamel saliva-coated slabs of known surface hardness (SH) and immersed in culture media. Biofilms were exposed 8 times per day to 10% sucrose. Medium pH was monitored twice daily as a biofilm acidogenicity indicator. After 96 h, biofilms were collected to determine biomass and bacteria viability. Slab demineralization was calculated as percentage of SH loss (%SHL). Additionally, the model was tested with different concentrations of the initial inoculum (103, 106, 108 cells/ml) and different adhesion times (2 or 8 h). The dual-species biofilm revealed no LB effects on SM cariogenicity, without changes in acidogenicity or %SHL among groups (p > 0.05, n = 12). Lack of activity of LB on SM cariogenicity persisted even when 105 times higher concentration of the probiotic was tested. Coaggregation was not observed. In conclusion, findings suggest that LB does not reduce cariogenicity of SM in a validated experimental caries model.

Download Full-text

Analysis of cariogenic potential of alternative milk beverages by in vitro Streptococcus mutans biofilm model and ex vivo caries model

Archives of Oral Biology ◽

10.1016/j.archoralbio.2019.05.033 ◽

2019 ◽

Vol 105 ◽

pp. 52-58 ◽

Cited By ~ 3

Author(s):

Yan Huang ◽

Tatyana Thompson ◽

Yapin Wang ◽

Qingzhao Yu ◽

Lin Zhu ◽

...

Keyword(s):

Streptococcus Mutans ◽

Ex Vivo ◽

Biofilm Model ◽

Caries Model

Download Full-text

Differences in pH fall, phosphorus content and dissolution of enamel in layers of the oral bacterium Streptococcus mutans deposited in vitro on bovine enamel granules with and without fluoride varnish

Archives of Oral Biology ◽

10.1016/0003-9969(82)90003-6 ◽

1982 ◽

Vol 27 (12) ◽

pp. 1003-1006 ◽

Cited By ~ 6

Author(s):

H. Luoma ◽

A-R. Luoma

Keyword(s):

Streptococcus Mutans ◽

Phosphorus Content ◽

Fluoride Varnish ◽

Oral Bacterium ◽

Bovine Enamel

Download Full-text

THE EFFECT OF CURCUMA XANTHORRHIZA ETHANOL EXTRACT ON THE VIABILITY OF STREPTOCOCCUS MUTANS AND AGGREGATIBACTER ACTINOMYCETEMCOMITANS (DENTAL BIOFILM RESEARCH: IN VITRO STUDY)

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2017.v10s5.23087 ◽

2017 ◽

Vol 10 (17) ◽

pp. 30

Author(s):

Royan Diana ◽

Hedijanti Joenoes ◽

Ariadna A Djais

Keyword(s):

Streptococcus Mutans ◽

In Vitro Study ◽

Ethanol Extract ◽

Single Species ◽

Aggregatibacter Actinomycetemcomitans ◽

Vitro Study ◽

Dental Biofilm ◽

Significant Difference ◽

Curcuma Xanthorrhiza

Objective: This study aimed to compare the effect of Curcuma xanthrorrhiza ethanol extract to the viability of Streptococcus mutans and Aggregatibacter actinomycetemcomitans using single- and dual-species biofilm at different phases of formation.Methods: Biofilm models were incubated for 4, 12, and 24 hrs, then exposed to the extract at a concentration of 0.525%.Results: The viability of the single-species S. mutans biofilm was low (p<0.05), and no significant difference (p>0.05) was found between singlespeciesA. actinomycetemcomitans and dual-species biofilm.Conclusions: Curcuma xanthorrhiza ethanol extract is more effective for decreasing the viability of single-species S. mutans biofilm.

Download Full-text

Preparation, characterisation and microbiological examination of Pickering nano-emulsions containing essential oils, and their effect on Streptococcus mutans biofilm treatment

Scientific Reports ◽

10.1038/s41598-019-52998-6 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Barbara Horváth ◽

Viktória L. Balázs ◽

Adorján Varga ◽

Andrea Böszörményi ◽

Béla Kocsis ◽

...

Keyword(s):

Essential Oils ◽

Streptococcus Mutans ◽

Dental Health ◽

Model Membrane ◽

Biofilm Model ◽

Mouth Care ◽

Modified Silica Nanoparticles ◽

Surface Modified ◽

20 Nm

Abstract Essential oils (EOs) are commonly applied in mouth care products like mouthwashes, mostly as an ethanolic solution or by usage of surfactants as solubilising agents. In this study, we present a formulation for preparation of Pickering nano-emulsions (PnE) of EOs as a novel form for application of EOs in mouth care. For the preparation of PnE, we have synthesised surface-modified silica nanoparticles with a mean diameter of 20 nm, as well as we have examined the effect of EOs concentration on PnE droplet size and stability. In vitro study of their effect on the Streptococcus mutans biofilm as the main pathogen of dental health problems has been performed. We have found that EOs in the PnE form has the highest effectiveness against biofilm formation. Diffusion through the biofilm model membrane was studied to explain this observation. We have found that PnEs have a better performance in the transportation of EOs trough model membrane than the ethanolic solutions and conventional emulsions (CEs).

Download Full-text

Probiotic Effects on Multispecies Biofilm Composition, Architecture, and Caries Activity In Vitro

Microorganisms ◽

10.3390/microorganisms8091272 ◽

2020 ◽

Vol 8 (9) ◽

pp. 1272

Author(s):

Zhihui Chen ◽

Sebastian Schlafer ◽

Gerd Göstemeyer ◽

Falk Schwendicke

Keyword(s):

Lactobacillus Reuteri ◽

Quantitative Polymerase Chain Reaction ◽

Lactobacillus Rhamnosus ◽

Negative Control ◽

Caries Activity ◽

Actinomyces Naeslundii ◽

Colony Forming Units ◽

Bovine Enamel ◽

Culture Supernatants

While probiotics have been tested for their anti-caries effect in vitro and also clinically, there is a lack of understanding of their effects on complex dental biofilms. We assessed two probiotics, Lactobacillus reuteri and Streptococcus oligofermentans, on a continuous-cultured model containing Streptococcus mutans, Lactobacillus rhamnosus and Actinomyces naeslundii. Cariogenic biofilms were grown on bovine enamel specimens and daily challenged with L. reuteri or S. oligofermentans whole culture (LC/SC) or cell-free supernatant (LS/SS) or medium only (negative control, NC) (n = 21/group) for 10 days. Biofilm was assessed via counting colony-forming units, quantitative polymerase chain reaction, and fluorescence in situ hybridization. Caries activity was determined by pH measurements and by assessing mineral loss (ΔZ) using transverse microradiography. Both LC and SC significantly reduced total and strain-specific cariogenic bacterial numbers (p < 0.05). ΔZ was reduced in LC (mean ± SD: 1846.67 ± 317.89) and SC (3315.87 ± 617.30) compared to NC (4681.48 ± 495.18, p < 0.05). No significant reductions in bacterial numbers and ΔZ was induced by supernatants. Biofilm architecture was not considerably affected by probiotic applications. Viable probiotics L. reuteri and S. oligofermentans, but not their culture supernatants, could reduce the caries activity of multi-species biofilms in vitro.

Download Full-text

Symbiotic Relationship between Streptococcus mutans and Candida albicans Synergizes Virulence of Plaque BiofilmsIn Vivo

Infection and Immunity ◽

10.1128/iai.00087-14 ◽

2014 ◽

Vol 82 (5) ◽

pp. 1968-1981 ◽

Cited By ~ 216

Author(s):

Megan L. Falsetta ◽

Marlise I. Klein ◽

Punsiri M. Colonne ◽

Kathleen Scott-Anne ◽

Stacy Gregoire ◽

...

Keyword(s):

Candida Albicans ◽

Dental Caries ◽

Streptococcus Mutans ◽

Bacterial Pathogen ◽

Single Species ◽

Content Type ◽

3 Dimensional ◽

Biofilm Development

ABSTRACTStreptococcus mutansis often cited as the main bacterial pathogen in dental caries, particularly in early-childhood caries (ECC).S. mutansmay not act alone;Candida albicanscells are frequently detected along with heavy infection byS. mutansin plaque biofilms from ECC-affected children. It remains to be elucidated whether this association is involved in the enhancement of biofilm virulence. We showed that the ability of these organisms together to form biofilms is enhancedin vitroandin vivo. The presence ofC. albicansaugments the production of exopolysaccharides (EPS), such that cospecies biofilms accrue more biomass and harbor more viableS. mutanscells than single-species biofilms. The resulting 3-dimensional biofilm architecture displays sizeableS. mutansmicrocolonies surrounded by fungal cells, which are enmeshed in a dense EPS-rich matrix. Using a rodent model, we explored the implications of this cross-kingdom interaction for the pathogenesis of dental caries. Coinfected animals displayed higher levels of infection and microbial carriage within plaque biofilms than animals infected with either species alone. Furthermore, coinfection synergistically enhanced biofilm virulence, leading to aggressive onset of the disease with rampant carious lesions. Ourin vitrodata also revealed that glucosyltransferase-derived EPS is a key mediator of cospecies biofilm development and that coexistence withC. albicansinduces the expression of virulence genes inS. mutans(e.g.,gtfB,fabM). We also found thatCandida-derived β1,3-glucans contribute to the EPS matrix structure, while fungal mannan and β-glucan provide sites for GtfB binding and activity. Altogether, we demonstrate a novel mutualistic bacterium-fungus relationship that occurs at a clinically relevant site to amplify the severity of a ubiquitous infectious disease.

Download Full-text

Impact of an Anticaries Mouthrinse onIn VitroRemineralization and Microbial Control

International Journal of Dentistry ◽

10.1155/2014/982071 ◽

2014 ◽

Vol 2014 ◽

pp. 1-11 ◽

Cited By ~ 2

Author(s):

Frank C. Sun ◽

E. Eric Engelman ◽

James A. McGuire ◽

Gabrielle Kosmoski ◽

Lauren Carratello ◽

...

Keyword(s):

Antimicrobial Agents ◽

Microbial Control ◽

Single Species ◽

Surface Microhardness ◽

Fluoride Uptake ◽

Biofilm Model ◽

Ph Cycling ◽

Flow Through ◽

Biofilm Matrix

Objective. The objective of this research was to evaluate the caries control potential of a new fluoride mouthrinse that also contained antimicrobial agents and a biofilm disrupting agent using differentin vitromodels.Methods. Fourin vitrostudies were conducted to assess the performance of this three pronged approach to caries control: (1) traditional enamel fluoride uptake, (2) surface microhardness study using pH cycling model and subsequent fluoride uptake, (3) a salivary biofilm flow-through study to determine the anti-microbial activity, and (4) a single species biofilm model measuring effect on biofilm matrix disruption.Results. The data showed that a LISTERINE rinse with fluoride, essential oils and xylitol was superior in promoting enamel fluoride uptake and in enhancing antimicrobial activity over traditional commercially available fluoridated products. An increase of the surface microhardness was observed when the LISTERINE rinse was used in combination with fluoridated toothpaste versus the fluoridated toothpaste alone. Finally, it was demonstrated that xylitol solutions disrupted and reduced the biovolume of biofilm matrix of matureStreptococcus mutans.Conclusion. Thesein vitrostudies demonstrated that a fluoride mouthrinse with antimicrobial agent and biofilm matrix disrupting agent provided multifaceted and enhanced anti-caries efficacy by promoting remineralization, reducing acidogenic bacteria and disrupting biofilm matrix.

Download Full-text

Cariogenic Potential of Human and Bovine Milk on Enamel Demineralization

Caries Research ◽

10.1159/000516090 ◽

2021 ◽

pp. 1-8

Author(s):

Antonio P. Ricomini Filho ◽

Ana Camila M. de Assis ◽

Bárbara E. Costa Oliveira ◽

Jaime A. Cury

Keyword(s):

Human Milk ◽

Bovine Milk ◽

Surface Hardness ◽

Human Saliva ◽

Positive Control ◽

Negative Control ◽

Extracellular Polysaccharides ◽

Enamel Demineralization ◽

Biofilm Model ◽

Bovine Enamel

The higher cariogenicity of human milk when compared with bovine milk is still a debatable subject. Therefore, we evaluated the effect of human or bovine milk exposure on biofilm composition and enamel demineralization using a validated cariogenic biofilm model. Streptococcus mutans UA159 biofilms (n = 8) were grown on human saliva-coated bovine enamel slabs of known surface hardness. The biofilms were exposed 8×/day to 0.9% NaCl (negative control), human milk, bovine milk, 7.0% lactose (active human milk control), 4.5% lactose (active bovine milk control), or 10% sucrose (positive control). The culture medium was changed twice daily, and the pH was analyzed as an indicator of biofilm acidogenicity. After 120 h of growth, biofilms were harvested to evaluate viable cells, and soluble and insoluble extracellular polysaccharides (EPS). Enamel demineralization was assessed by the percentage of surface hardness loss (%SHL). Data were analyzed by one-way ANOVA/Tukey’s test (α = 5%). In terms of %SHL, negative control (7.7 ± 3.1), human milk control (13.3 ± 7.5), bovine milk control (15.3 ± 8.2), human milk (7.5 ± 5.0), and bovine milk (8.7 ± 6.3) did not differ among them (p > 0.05) but differed (p < 0.05) from sucrose (55.1 ± 5.4). The findings of enamel demineralization (%SHL) were statistically supported by the data of biofilm acidogenicity, bacterial counts and EPS biofilm composition. This experimental study suggests that human and bovine milk have low cariogenic potential to provoke caries lesions in enamel.

Download Full-text