Inhibited biofilm formation and improved antibacterial activity of a novel nanoemulsion against cariogenic Streptococcus mutans in vitro and in vivo

Streptococcus mutans is the major cause of dental plaque and is often associated with biofilm formation. The aim of this study is to evaluate the activity of a hydrosoluble derivative of chitosan against S. mutans biofilms in vitro and in vivo. Strains of S. mutans were isolated from the dental plaque of 84 patients enrolled in the study. The antibacterial activity of chitosan was determined by broth microdilutions. The effect of chitosan at different concentrations and exposure times on S. mutans biofilms at different phases of development was assessed by a clinical study using the classical “4-day plaque regrowth” experiment in adult volunteers. The MIC values of chitosan were between 0.5 and 2 g/L. Compared to distilled water, the chitosan solution significantly decreased the vitality of plaque microflora (p≤0.05). Chlorhexidine, used as a positive control, reduced vitality even further. The results showed that S. mutans in the adhesion phase (4 h) was completely inhibited by chitosan at any concentration (0.1, 0.2, 0.5XMIC) or exposure time investigated (1, 15, 30, 60 min), while S. mutans at successive stages of accumulation (12–24 h) was inhibited only by higher concentrations and longer exposure times. These data confirm the effective action of chitosan against S. mutans biofilms.

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Molecule Targeting Glucosyltransferase Inhibits Streptococcus mutans Biofilm Formation and Virulence

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00919-15 ◽

2015 ◽

Vol 60 (1) ◽

pp. 126-135 ◽

Cited By ~ 44

Author(s):

Zhi Ren ◽

Tao Cui ◽

Jumei Zeng ◽

Lulu Chen ◽

Wenling Zhang ◽

...

Keyword(s):

Streptococcus Mutans ◽

Dental Plaque ◽

Biofilm Formation ◽

Extracellular Polysaccharides ◽

Content Type ◽

Oral Infections ◽

Concomitant Reduction ◽

New Strategies

ABSTRACTDental plaque biofilms are responsible for numerous chronic oral infections and cause a severe health burden. Many of these infections cannot be eliminated, as the bacteria in the biofilms are resistant to the host's immune defenses and antibiotics. There is a critical need to develop new strategies to control biofilm-based infections. Biofilm formation inStreptococcus mutansis promoted by major virulence factors known as glucosyltransferases (Gtfs), which synthesize adhesive extracellular polysaccharides (EPS). The current study was designed to identify novel molecules that target Gtfs, thereby inhibitingS. mutansbiofilm formation and having the potential to prevent dental caries. Structure-based virtual screening of approximately 150,000 commercially available compounds against the crystal structure of the glucosyltransferase domain of the GtfC protein fromS. mutansresulted in the identification of a quinoxaline derivative, 2-(4-methoxyphenyl)-N-(3-{[2-(4-methoxyphenyl)ethyl]imino}-1,4-dihydro-2-quinoxalinylidene)ethanamine, as a potential Gtf inhibitor.In vitroassays showed that the compound was capable of inhibiting EPS synthesis and biofilm formation inS. mutansby selectively antagonizing Gtfs instead of by killing the bacteria directly. Moreover, thein vivoanti-caries efficacy of the compound was evaluated in a rat model. We found that the compound significantly reduced the incidence and severity of smooth and sulcal-surface cariesin vivowith a concomitant reduction in the percentage ofS. mutansin the animals' dental plaque (P< 0.05). Taken together, these results represent the first description of a compound that targets Gtfs and that has the capacity to inhibit biofilm formation and the cariogenicity ofS. mutans.

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Silver-Containing Hydroxyapatite Coating Reduces Biofilm Formation by Methicillin-ResistantStaphylococcus aureusIn Vitro and In Vivo

BioMed Research International ◽

10.1155/2016/8070597 ◽

2016 ◽

Vol 2016 ◽

pp. 1-7 ◽

Cited By ~ 11

Author(s):

Masaya Ueno ◽

Hiroshi Miyamoto ◽

Masatsugu Tsukamoto ◽

Shuichi Eto ◽

Iwao Noda ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Antibacterial Activity ◽

Biofilm Formation ◽

Thermal Spraying ◽

Time Points ◽

High Antibacterial Activity ◽

Ha Coating ◽

Coverage Rates

Biofilm-producing bacteria are the principal causes of infections associated with orthopaedic implants. We previously reported that silver-containing hydroxyapatite (Ag-HA) coatings exhibit high antibacterial activity against methicillin-resistantStaphylococcus aureus(MRSA). In the present study, we evaluated the effects of Ag-HA coating of implant surfaces on biofilm formation. Titanium disks (14-mm diameter, 1-mm thickness), one surface of which was coated with HA or 0.5%–3.0% Ag-HA with a thermal spraying technique, were used. In vitro, the disks were inoculated with an MRSA suspension containing4×105 CFU and incubated for 1-2 weeks. In vivo, MRSA-inoculated HA and 3% Ag-HA disks (8.8–10.0 × 108 CFU) were implanted subcutaneously on the back of rats for 1–7 days. All disks were subsequently stained with a biofilm dye and observed under a fluorescence microscope, and biofilm coverage rates (BCRs) were calculated. The BCRs on the Ag-HA coating were significantly lower than those on the HA coating at all time points in vitro (p<0.05). Similar results were observed in vivo (p<0.001) without argyria. Ag-HA coating reduced biofilm formation by MRSA in vitro and in vivo; therefore, Ag-HA coating might be effective for reducing implant-associated infections.

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Reduction of Streptococcus mutans Adherence and Dental Biofilm Formation by Surface Treatment with Phosphorylated Polyethylene Glycol

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00380-07 ◽

2007 ◽

Vol 51 (10) ◽

pp. 3634-3641 ◽

Cited By ~ 27

Author(s):

Akira Shimotoyodome ◽

Takashi Koudate ◽

Hisataka Kobayashi ◽

Junji Nakamura ◽

Ichiro Tokimitsu ◽

...

Keyword(s):

Polyethylene Glycol ◽

Streptococcus Mutans ◽

Biofilm Formation ◽

De Novo ◽

Bacterial Attachment ◽

Salivary Proteins ◽

Dental Biofilm ◽

Biofilm Development

ABSTRACT Initial attachment of the cariogenic Streptococcus mutans onto dental enamel is largely promoted by the adsorption of specific salivary proteins on enamel surface. Some phosphorylated salivary proteins were found to reduce S. mutans adhesion by competitively inhibiting the adsorption of S. mutans-binding salivary glycoproteins to hydroxyapatite (HA). The aim of this study was to develop antiadherence compounds for preventing dental biofilm development. We synthesized phosphorylated polyethylene glycol (PEG) derivatives and examined the possibility of surface pretreatment with them for preventing S. mutans adhesion in vitro and dental biofilm formation in vivo. Pretreatment of the HA surface with methacryloyloxydecyl phosphate (MDP)-PEG prior to saliva incubation hydrophilized the surface and thereby reduced salivary protein adsorption and saliva-promoted bacterial attachment to HA. However, when MDP-PEG was added to the saliva-pretreated HA (S-HA) surface, its inhibitory effect on bacterial binding was completely diminished. S. mutans adhesion onto S-HA was successfully reduced by treatment of the surface with pyrophosphate (PP), which desorbs salivary components from S-HA. Treatment of S-HA surfaces with MDP-PEG plus PP completely inhibited saliva-promoted S. mutans adhesion even when followed by additional saliva treatment. Finally, mouthwash with MDP-PEG plus PP prevented de novo biofilm development after thorough teeth cleaning in humans compared to either water or PP alone. We conclude that MDP-PEG plus PP has the potential for use as an antiadherence agent that prevents dental biofilm development.

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The Anti-Biofilm Activity of Oregano Essential Oil Against Dental Plaque-Forming Streptococcus mutans In Vitro and In Vivo

Journal of Kermanshah University of Medical Sciences ◽

10.5812/jkums.107680 ◽

2020 ◽

Vol 24 (3) ◽

Author(s):

Fatemeh Hejazinia ◽

Leila Fozouni ◽

Nasrin Sadat Azami ◽

Seyedgholamreza Mousavi

Keyword(s):

Essential Oil ◽

Elementary School Students ◽

Streptococcus Mutans ◽

Biofilm Formation ◽

Microtiter Plate ◽

School Students ◽

Microtiter Plate Assay ◽

Oregano Essential Oil

Background: The oral and dental infections that are mainly caused by bacterial biofilms are among the most prevalent human infections worldwide. Objectives: The present study aimed to investigate the in vitro and in vivo inhibitory and anti-biofilm effects of oregano essential oil on the Streptococcus mutans isolates obtained from elementary school students. Methods: This experimental study was conducted on 150 samples collected from the buccal and lingual surfaces of the posterior teeth of elementary school students. S. mutans strains were identified using conventional microbiological and biochemical tests, and biofilm formation was assessed using the microtiter plate assay. The minimum inhibitory concentration (MIC) of the oregano essential oil against the isolates was determined using the broth microdilution method. In addition, the effective constituents of the essential oil were measured via gas chromatography-mass spectroscopy. The in vitro and in vivo anti-biofilm activities of the oregano essential oil were also evaluated using the modified microtiter plate assay and on the tooth surfaces of male NMRI mice, respectively. Results: The frequency of S. mutans was 15.3%, 87% of which were capable of biofilm formation. The MIC of the oregano essential oil was 50 µl/ml against the S. mutans isolates, and 82% of the isolates did not grow at the concentrations of ≥ 512 µl/ml. However, none of the isolates were capable of biofilm formation at the MIC and sub-MIC concentrations of the essential oil. Limonene and myrcene were the most effective constituents of the essential oil. Furthermore, a significant correlation was observed between treatment with the oregano essential oil and biofilm formation by the streptococci isolates (P = 0.05). Conclusions: According to the results, the presence of biofilm and incidence of dental caries were significantly correlated. Moreover, the essential oil of oregano and its main constituents had potent anti-biofilm and antibacterial properties and could be utilized for the production of new plant-based mouthwashes.

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PspC domain-containing protein (PCP) determines Streptococcus mutans biofilm formation through bacterial extracellular DNA release and platelet adhesion in experimental endocarditis

PLoS Pathogens ◽

10.1371/journal.ppat.1009289 ◽

2021 ◽

Vol 17 (2) ◽

pp. e1009289

Author(s):

Chiau-Jing Jung ◽

Chih-Chieh Hsu ◽

Jeng-Wei Chen ◽

Hung-Wei Cheng ◽

Chang-Tsu Yuan ◽

...

Keyword(s):

Streptococcus Mutans ◽

Biofilm Formation ◽

Heart Valves ◽

Platelet Adhesion ◽

Bacterial Biofilm ◽

Extracellular Dna ◽

Dependent Manner ◽

Dna Release

Bacterial extracellular DNA (eDNA) and activated platelets have been found to contribute to biofilm formation by Streptococcus mutans on injured heart valves to induce infective endocarditis (IE), yet the bacterial component directly responsible for biofilm formation or platelet adhesion remains unclear. Using in vivo survival assays coupled with microarray analysis, the present study identified a LiaR-regulated PspC domain-containing protein (PCP) in S. mutans that mediates bacterial biofilm formation in vivo. Reverse transcriptase- and chromatin immunoprecipitation-polymerase chain reaction assays confirmed the regulation of pcp by LiaR, while PCP is well-preserved among streptococcal pathogens. Deficiency of pcp reduced in vitro and in vivo biofilm formation and released the eDNA inside bacteria floe along with reduced bacterial platelet adhesion capacity in a fibrinogen-dependent manner. Therefore, LiaR-regulated PCP alone could determine release of bacterial eDNA and binding to platelets, thus contributing to biofilm formation in S. mutans-induced IE.

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The Role of Candida albicans Secreted Polysaccharides in Augmenting Streptococcus mutans Adherence and Mixed Biofilm Formation: In vitro and in vivo Studies

Frontiers in Microbiology ◽

10.3389/fmicb.2020.00307 ◽

2020 ◽

Vol 11 ◽

Cited By ~ 3

Author(s):

Zaid H. Khoury ◽

Taissa Vila ◽

Taanya R. Puthran ◽

Ahmed S. Sultan ◽

Daniel Montelongo-Jauregui ◽

...

Keyword(s):

Candida Albicans ◽

Streptococcus Mutans ◽

Biofilm Formation ◽

In Vivo Studies

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Evaluation of Physical Parameters of Novel Licorice Varnish Versus Fluoride and Combination Varnish: An In-Vitro Study

Acta Medica Academica ◽

10.5644/ama2006-124.229 ◽

2018 ◽

Vol 47 (2) ◽

pp. 176

Author(s):

Roopali Sankeshwari ◽

Anil Ankola ◽

Kishore Bhat ◽

Udaya Bolmal ◽

Malleswara Rao

Keyword(s):

Antibacterial Activity ◽

Shelf Life ◽

Streptococcus Mutans ◽

Brain Heart Infusion ◽

Tooth Surface ◽

Physical Parameters ◽

Activity Assessment ◽

Rate Of Evaporation

Objectives. The aim of this study was to evaluate the physical properties of locally prepared Licorice varnish (LV), commercially available Fluoride varnish (FV) and a Combination of both Varnishes (CV).Material and Methods. LV was prepared using authenticated licorice roots. Commercially available FV (Bifluorid 12) was used as a positive control and CV was prepared in six different concentrations of both varnishes. Conventional antibacterial activity assessment, employing disc diffusion and broth dilution methods, was inconclusive. Therefore a novel assessment method was used, whereby the varnish was directly added to a mixture of Brain Heart Infusion broth with Streptococcus mutans and incubated. Physical parameters such as pH, rate of evaporation, viscosity, film forming ability, and cost incurred for preparation were assessed and compared.Results. FV, LV and CV (except the combination of LV 80% + FV 20%) showed antibacterial activity against Streptococcus mutans. All three varnishes formed films on the tooth surface as confirmed by Scanning Electron Microscopy. Mean pH was in the range of 4-4.5, viscosity 48-52 centipoise (cP), rate of evaporation was 150-160 seconds. They were comparable to each other in the physical parameters tested, except for the shelf life of LV.Conclusion. All three varnishes showed antibacterial activity against Streptococcus mutans which was established using an innovative method of antibacterial activity assessment. LV was most economical of all but had a shorter shelf life. The results of this study need to be evaluated through an in vivo study.

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Multi-Omics Reveals the Inhibition of Lactiplantibacillus plantarum CCFM8724 in Streptococcus mutans-Candida albicans Mixed-Species Biofilms

Microorganisms ◽

10.3390/microorganisms9112368 ◽

2021 ◽

Vol 9 (11) ◽

pp. 2368

Author(s):

Qiuxiang Zhang ◽

Jiaxun Li ◽

Wenwei Lu ◽

Jianxin Zhao ◽

Hao Zhang ◽

...

Keyword(s):

Gene Expression ◽

Candida Albicans ◽

Dental Caries ◽

Streptococcus Mutans ◽

Biofilm Formation ◽

Metabolic Reprogramming ◽

Mixed Species ◽

Carbohydrate Utilization

Lactiplantibacillus plantarum CCFM8724 is a probiotic with the potential to prevent dental caries in vitro and in vivo. To explore the effects of this probiotic at inhibiting Streptococcus mutans-Candida albicans mixed-species biofilm and preventing dental caries, multi-omics, including metabolomics and transcriptomics, was used to investigate the regulation of small-molecule metabolism during biofilm formation and the gene expression in the mixed-species biofilm. Metabolomic analysis revealed that some carbohydrates related to biofilm formation, such as sucrose, was detected at lower levels due to the treatment with the L. plantarum supernatant. Some sugar alcohols, such as xylitol and sorbitol, were detected at higher levels, which may have inhibited the growth of S. mutans. In transcriptomic analysis, the expression of the virulence genes of C. albicans, such as those that code agglutinin-like sequence (Als) proteins, was affected. In addition, metabolomics coupled with a Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and RNA-seq revealed that the L. plantarum supernatant had an active role in sugar metabolism during the formation of the S. mutans-C. albicans mixed-species biofilm, and the L. plantarum supernatant was also related to carbohydrate utilization, glucan biosynthesis, and mycelium formation. Hence, L. plantarum CCFM8724 decreased the mixed-species biofilm mass from the perspective of gene expression and metabolic reprogramming. Our results provide a rationale for evaluating L. plantarum CCFM8724 as a potential oral probiotic for inhibiting cariogenic pathogen biofilm formation and improving dental caries.

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