Mediterranean herb extracts inhibit microbial growth of representative oral microorganisms and biofilm formation of Streptococcus mutans

Several new products with innovative formulations are being proposed to facilitate oral care. Here, we evaluated the effects of a commercially available product, a toothpaste and chewing gum named Biorepair Peribioma, on oral microorganisms of healthy subjects. Saliva from six volunteers was collected during 20 min of mastication of a traditional gum (gum A) and the Biorepair Peribioma gum (gum P). Orthodontic elastics (OE) were in vitro contaminated with salivary samples, both A and P, and subsequently exposed or not to a Biorepair Peribioma toothpaste-conditioned supernatant (Tp-SUP). The salivary samples were tested for initial microbial load; hence, the contaminated OE were assessed for microbial growth, adhesion, biofilm formation and persistence; moreover, species identification was assessed. We found that the salivary samples A and P had similar microbial load; upon contamination, microbial adhesion onto the OE was detected to a lower extent when using saliva P with respect to saliva A. Microbial growth and biofilm formation, assessed at 24 h, remained at lower levels in OE exposed to saliva P, compared to saliva A. This difference between salivary samples A and P was confirmed when measuring biofilm persistence (48 h), while it was lost in terms of microbial re-growth (48 h). The Tp-SUP treatment drastically affected microbial load at 24 h and strongly impaired biofilm formation/persistence, in OE exposed to both salivary samples A and P. Finally, such treatment resulted in consistent overgrowth of Lactobacilli, bacterial species originally present both in the Biorepair Peribioma toothpaste and gum. In conclusion, by an in vitro pilot study, we show that the Biorepair Peribioma toothpaste and gum deeply affect oral microorganisms’ behavior, drastically impairing their ability to contaminate and produce plaque onto orthodontic devices.

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Pu'er tea rich in strictinin and catechins prevents biofilm formation of two cariogenic bacteria, Streptococcus mutans and Streptococcus sobrinus

Journal of Dental Sciences ◽

10.1016/j.jds.2021.05.011 ◽

2021 ◽

Author(s):

Man-Hua Liao ◽

Xiu-Ru Wang ◽

Wei-Li Hsu ◽

Jason T.C. Tzen

Keyword(s):

Streptococcus Mutans ◽

Biofilm Formation ◽

Streptococcus Sobrinus ◽

Cariogenic Bacteria

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Trk2 Potassium Transport System in Streptococcus mutans and Its Role in Potassium Homeostasis, Biofilm Formation, and Stress Tolerance

Journal of Bacteriology ◽

10.1128/jb.00813-15 ◽

2016 ◽

Vol 198 (7) ◽

pp. 1087-1100 ◽

Cited By ~ 14

Author(s):

Gursonika Binepal ◽

Kamal Gill ◽

Paula Crowley ◽

Martha Cordova ◽

L. Jeannine Brady ◽

...

Keyword(s):

Stress Tolerance ◽

Streptococcus Mutans ◽

Transport System ◽

Biofilm Formation ◽

Cellular Response ◽

Pathogenic Bacteria ◽

Transport Systems ◽

Content Type ◽

Growth And Survival ◽

Dental Biofilm

ABSTRACTPotassium (K+) is the most abundant cation in the fluids of dental biofilm. The biochemical and biophysical functions of K+and a variety of K+transport systems have been studied for most pathogenic bacteria but not for oral pathogens. In this study, we establish the modes of K+acquisition inStreptococcus mutansand the importance of K+homeostasis for its virulence attributes. TheS. mutansgenome harbors four putative K+transport systems that included two Trk-like transporters (designated Trk1 and Trk2), one glutamate/K+cotransporter (GlnQHMP), and a channel-like K+transport system (Kch). Mutants lacking Trk2 had significantly impaired growth, acidogenicity, aciduricity, and biofilm formation. [K+] less than 5 mM eliminated biofilm formation inS. mutans. The functionality of the Trk2 system was confirmed by complementing anEscherichia coliTK2420 mutant strain, which resulted in significant K+accumulation, improved growth, and survival under stress. Taken together, these results suggest that Trk2 is the main facet of the K+-dependent cellular response ofS. mutansto environment stresses.IMPORTANCEBiofilm formation and stress tolerance are important virulence properties of caries-causingStreptococcus mutans. To limit these properties of this bacterium, it is imperative to understand its survival mechanisms. Potassium is the most abundant cation in dental plaque, the natural environment ofS. mutans. K+is known to function in stress tolerance, and bacteria have specialized mechanisms for its uptake. However, there are no reports to identify or characterize specific K+transporters inS. mutans. We identified the most important system for K+homeostasis and its role in the biofilm formation, stress tolerance, and growth. We also show the requirement of environmental K+for the activity of biofilm-forming enzymes, which explains why such high levels of K+would favor biofilm formation.

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A Novel Gene Involved in the Survival of Streptococcus mutans under Stress Conditions

Applied and Environmental Microbiology ◽

10.1128/aem.02549-13 ◽

2013 ◽

Vol 80 (1) ◽

pp. 97-103 ◽

Cited By ~ 11

Author(s):

Dan Li ◽

Yukie Shibata ◽

Toru Takeshita ◽

Yoshihisa Yamashita

Keyword(s):

Streptococcus Mutans ◽

Streptococcus Pyogenes ◽

Biofilm Formation ◽

Acid Tolerance ◽

Stress Conditions ◽

Insertion Mutagenesis ◽

Trna Modification ◽

Content Type ◽

Virulence Proteins ◽

Random Insertion

ABSTRACTAStreptococcus mutansmutant defective in aciduricity was constructed by random-insertion mutagenesis. Sequence analysis of the mutant revealed a mutation ingidA, which is known to be involved in tRNA modification inStreptococcus pyogenes. Complementation ofgidAbyS. pyogenesgidArecovered the acid tolerance ofS. mutans. Although thegidA-inactivatedS. pyogenesmutant exhibited significantly reduced expression of multiple extracellular virulence proteins, theS. mutansmutant did not. On the other hand, thegidAmutant ofS. mutansshowed reduced ability to withstand exposure to other stress conditions (high osmotic pressure, high temperature, and bacitracin stress) besides an acidic environment. In addition, loss of GidA decreased the capacity for glucose-dependent biofilm formation by over 50%. This study revealed thatgidAplays critical roles in the survival ofS. mutansunder stress conditions, including lower pH.

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Effect of a Berry Polyphenolic Fraction on Biofilm Formation, Adherence Properties and Gene Expression of Streptococcus mutans and Its Biocompatibility with Oral Epithelial Cells

Antibiotics ◽

10.3390/antibiotics10010046 ◽

2021 ◽

Vol 10 (1) ◽

pp. 46

Author(s):

Mariem Souissi ◽

Amel Ben Lagha ◽

Kamel Chaieb ◽

Daniel Grenier

Keyword(s):

Epithelial Cells ◽

Streptococcus Mutans ◽

Biofilm Formation ◽

Cell Model ◽

Pcr Analysis ◽

Adhesion Properties ◽

Oral Epithelial Cells ◽

Polymerase Chain ◽

Oral Epithelial ◽

Wild Blueberry

The ability of Streptococcus mutans to adhere to oral surfaces and form biofilm is a key step in the tooth decay process. The aim of this study was to investigate a berry (wild blueberry, cranberry, and strawberry) polyphenolic fraction, commercialized as Orophenol®, for its antibacterial, anti-biofilm, and anti-adhesion properties on S. mutans. Moreover, the biocompatibility of the fraction with human oral epithelial cells was assessed. Phenolic acids, flavonoids (flavonols, anthocyanins, flavan-3-ols), and procyanidins made up 10.71%, 19.76%, and 5.29% of the berry polyphenolic fraction, respectively, as determined by chromatography and mass spectrometry. The berry polyphenolic preparation dose-dependently inhibited S. mutans biofilm formation while not reducing bacterial growth. At concentrations ranging from 250 to 1000 µg/mL, the fraction inhibited the adhesion of S. mutans to both saliva-coated hydroxyapatite and saliva-coated nickel–chrome alloy. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis showed that incubating S. mutans with the berry polyphenolic fraction was associated with a reduced expression of luxS gene, which regulates quorum sensing in S. mutans. The berry fraction did not show any significant cytotoxicity in an oral epithelial cell model. In conclusion, Orophenol®, which is a mixture of polyphenols from wild blueberry, cranberry and strawberry, possesses interesting anti-caries properties while being compatible with oral epithelial cells.

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The Effects of Surface Roughness of Composite Resin on Biofilm Formation of Streptococcus mutans in the Presence of Saliva

Operative Dentistry ◽

10.2341/11-371-l ◽

2012 ◽

Vol 37 (5) ◽

pp. 532-539 ◽

Cited By ~ 36

Author(s):

JW Park ◽

CW Song ◽

JH Jung ◽

SJ Ahn ◽

JL Ferracane

Keyword(s):

Surface Roughness ◽

Silicon Carbide ◽

Streptococcus Mutans ◽

Biofilm Formation ◽

Composite Resin ◽

Resin Composite ◽

Fluid Phase ◽

Confocal Laser ◽

Carbohydrate Source ◽

Effect Of Surface

SUMMARY The purpose of this study was to investigate the effects of surface roughness of resin composite on biofilm formation of Streptococcus mutans in the presence of saliva. To provide uniform surface roughness on composites, disks were prepared by curing composite against 400-grit silicon carbide paper (SR400), 800-grit silicon carbide paper (SR800), or a glass slide (SRGlass). The surface roughness was examined using confocal laser microscopy. For biofilm formation, S. mutans was grown for 24 hours with each disk in a biofilm medium with either glucose or sucrose in the presence of fluid-phase or surface-adsorbed saliva. The adherent bacteria were quantified via enumeration of the total viable counts of bacteria. Biofilms were examined using scanning electron microscopy. This study showed that SR400 had deeper and larger, but fewer depressions than SR800. Compared to SRGlass and SR800, biofilm formation was significantly increased on SR400. In addition, the differences in the effect of surface roughness on the amount of biofilm formation were not significantly influenced by either the presence of saliva or the carbohydrate source. Considering that similar differences in surface roughness were observed between SR400 and SR800 and between SR800 and SRGlass, this study suggests that surface topography (size and depth of depressions) may play a more important role than surface roughness in biofilm formation of S. mutans.

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Effects of sub-minimum inhibitory concentrations of antimicrobial agents on Streptococcus mutans biofilm formation

International Journal of Antimicrobial Agents ◽

10.1016/j.ijantimicag.2012.01.009 ◽

2012 ◽

Vol 39 (5) ◽

pp. 390-395 ◽

Cited By ~ 47

Author(s):

Liping Dong ◽

Zhongchun Tong ◽

Dake Linghu ◽

Yuan Lin ◽

Rui Tao ◽

...

Keyword(s):

Streptococcus Mutans ◽

Biofilm Formation ◽

Antimicrobial Agents ◽

Minimum Inhibitory Concentrations

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Effects of pH on the Properties of Membrane Vesicles Including Glucosyltransferase in Streptococcus mutans

Microorganisms ◽

10.3390/microorganisms9112308 ◽

2021 ◽

Vol 9 (11) ◽

pp. 2308

Author(s):

Yusuke Iwabuchi ◽

Tomoyo Nakamura ◽

Yasuka Kusumoto ◽

Ryoma Nakao ◽

Tsutomu Iwamoto ◽

...

Keyword(s):

Streptococcus Mutans ◽

Biofilm Formation ◽

Membrane Vesicles ◽

Extracellular Dna ◽

Tooth Surface ◽

Initial Ph ◽

Effects Of Ph ◽

Low Levels ◽

Quantitative Changes ◽

Biofilm Development

Streptococcus mutans releases membrane vesicles (MVs) and induces MV-dependent biofilm formation. Glucosyltransferases (Gtfs) are bound to MVs and contribute to the adhesion and glucans-dependent biofilm formation of early adherent bacteria on the tooth surface. The biofilm formation of S. mutans may be controlled depending on whether the initial pH tends to be acidic or alkaline. In this study, the characteristics and effects of MVs extracted from various conditions {(initial pH 6.0 and 8.0 media prepared with lactic acid (LA) and acetic acid (AA), and with NaOH (NO), respectively)} on the biofilm formation of S. mutans and early adherent bacteria were investigated. The quantitative changes in glucans between primary pH 6.0 and 8.0 conditions were observed, associated with different activities affecting MV-dependent biofilm formation. The decreased amount of Gtfs on MVs under the initial pH 6.0 conditions strongly guided low levels of MV-dependent biofilm formation. However, in the initial pH 6.0 and 8.0 solutions prepared with AA and NO, the MVs in the biofilm appeared to be formed by the expression of glucans and/or extracellular DNA. These results suggest that the environmental pH conditions established by acid and alkaline factors determine the differences in the local pathogenic activities of biofilm development in the oral cavity.

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Low concentration of xylitol improves children tooth protection against Streptococcus mutans biofilm formation

Oral Health and Care ◽

10.15761/ohc.1000160 ◽

2019 ◽

Vol 4 (1) ◽

Cited By ~ 1

Author(s):

Dorota Kościelniak ◽

Iwona Gregorczyk-Maga ◽

Anna Jurczak ◽

Małgorzata Staszczyk ◽

Iwona Kołodziej ◽

...

Keyword(s):

Streptococcus Mutans ◽

Biofilm Formation ◽

Low Concentration

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Anti-Streptococcus mutans and anti-biofilm activities of dextranase and its encapsulation in alginate beads for application in toothpaste

PeerJ ◽

10.7717/peerj.10165 ◽

2020 ◽

Vol 8 ◽

pp. e10165

Author(s):

Nucharee Juntarachot ◽

Sasithorn Sirilun ◽

Duangporn Kantachote ◽

Phakkharawat Sittiprapaporn ◽

Piyachat Tongpong ◽

...

Keyword(s):

Sodium Alginate ◽

Calcium Chloride ◽

Streptococcus Mutans ◽

Dental Plaque ◽

Biofilm Formation ◽

Alginate Beads ◽

Oral Diseases ◽

Sensory Test ◽

Alginate Concentration ◽

The Stability

Background The accumulation of plaque causes oral diseases. Dental plaque is formed on teeth surfaces by oral bacterial pathogens, particularly Streptococcus mutans, in the oral cavity. Dextranase is one of the enzymes involved in antiplaque accumulation as it can prevent dental caries by the degradation of dextran, which is a component of plaque biofilm. This led to the idea of creating toothpaste containing dextranase for preventing oral diseases. However, the dextranase enzyme must be stable in the product; therefore, encapsulation is an attractive way to increase the stability of this enzyme. Methods The activity of food-grade fungal dextranase was measured on the basis of increasing ratio of reducing sugar concentration, determined by the reaction with 3, 5-dinitrosalicylic acid reagent. The efficiency of the dextranase enzyme was investigated based on its minimal inhibitory concentration (MIC) against biofilm formation by S. mutans ATCC 25175. Box-Behnken design (BBD) was used to study the three factors affecting encapsulation: pH, calcium chloride concentration, and sodium alginate concentration. Encapsulation efficiency (% EE) and the activity of dextranase enzyme trapped in alginate beads were determined. Then, the encapsulated dextranase in alginate beads was added to toothpaste base, and the stability of the enzyme was examined. Finally, sensory test and safety evaluation of toothpaste containing encapsulated dextranase were done. Results The highest activity of the dextranase enzyme was 4401.71 unit/g at a pH of 6 and 37 °C. The dextranase at its MIC (4.5 unit/g) showed strong inhibition against the growth of S. mutans. This enzyme at 1/2 MIC also showed a remarkable decrease in biofilm formation by S. mutans. The most effective condition of dextranase encapsulation was at a pH of 7, 20% w/v calcium chloride and 0.85% w/v sodium alginate. Toothpaste containing encapsulated dextranase alginate beads produced under suitable condition was stable after 3 months of storage, while the sensory test of the product was accepted at level 3 (like slightly), and it was safe. Conclusion This research achieved an alternative health product for oral care by formulating toothpaste with dextranase encapsulated in effective alginate beads to act against cariogenic bacteria, like S. mutants, by preventing dental plaque.

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