Estudio de la actividad antimicrobiana de Xenophyllum poposum sobre microorganismos de la cavidad bucal por el método de difusión radial / Study of the antimicrobial activity of Xenophyllum poposum on microorganisms of the oral cavity by the radial diffusion method

Resumen En este estudio se evaluó la actividad antimicrobiana in vitro de extractos de Xenophyllum poposum sobre microorganismos bucales como Streptococcus mutans, Streptococcus sobrinus, Lactobacillus casei, Actinomyces naeslundii, Actinomyces odontolyticus, Candida albicans y Veillonella sp. Se empleó el método de difusión radial en agar y como controles negativo y positivo de inhibición se emplearon etanol y clorhexidina al 0,12% (Plac out NF®) respectivamente. Los extractos con mayor actividad antimicrobiana fueron el etanólico y el clorofórmico. La diferencia entre ambos no fue estadísticamente significativa (p≥0,05). Tampoco se observó diferencia significativa con respecto a la clorhexidina, excepto sobre Veillonella sp., ya que el extracto etanólico presentó halos de inhibición significativamente menores sobre este microorganismo. Esto es importante ya que Veillonella se considera indicador de salud en relación a la caries dental. En base a esto, el extracto etanólico de Xenophyllum poposum podría ser usado como control químico de la biopelícula dental.

Abstract In this study, the in vitro antimicrobial activity of Xenophyllum poposum extracts on oral microorganisms such as Streptococcus mutans, Streptococcus sobrinus, Lactobacillus casei, Actinomyces naeslundii, Actinomyces odontolyticus, Candida albicans, Veillonella sp. was evaluated. The radial diffusion method in agar was used and 0.12% ethanol and chlorhexidine (Plac out NF®) were used as negative and positive inhibition controls, respectively. The extracts with the highest antimicrobial activity were the ethanolic and chloroform extracts. The difference between the two was not statistically significant (p≥0.05). No significant difference was observed with respect to chlorhexidine, except on Veillonella sp., since the ethanolic extract presented significantly lower inhibition halos on this microorganism. This is important as Veillonella is considered an indicator of health in relation to dental caries. Based on this, the ethanolic extract of Xenophyllum poposum could be used as chemical control of dental biofilm.

Resumo Neste estudo, a atividade antimicrobiana de extratos de Xenophyllum poposum sobre microrganismos orais como Streptococcus mutans, Streptococcus sobrinus, Lactobacillus casei, Actinomyces naeslundii, Actinomyces odontolyticus, Candida albicans e Veillonella sp. Foi utilizado o método de difusão radial em ágar e etanol 0,12% e clorexidina (Plac out NF®) como controles de inibição negativa e positiva, respectivamente. Os extratos com maior atividade antimicrobiana foram os extratos etanólico e clorofórmio. A diferença entre os dois não foi estatisticamente significativa (p≥0,05). Não foi observada diferença significativa em relação à clorexidina 0,12%, exceto em Veillonella sp., uma vez que o extrato etanólico apresentou halos de inibição significativamente menores neste microrganismo. Isso é importante, pois a Veillonella é considerada um indicador de saúde em relação à cárie dentária. Com base nisso, o extrato etanólico de Xenophyllum poposum pode ser utilizado como controle químico do biofilme dental.

Antimicrobial Photodynamic Treatment with Mother Juices and Their Single Compounds as Photosensitizers.

The potent antimicrobial effects of antimicrobial photodynamic therapy (aPDT) with visible light plus water-filtered infrared-A irradiation and natural compounds as photosensitizers (PSs) have recently been demonstrated. The aim of this study was to obtain information on the antimicrobial effects of aPDT with mother juices against typical cariogenic oral Streptococcus pathogens in their planktonic form and determine its eradication potential on total human salivary bacteria from volunteers. Mother juices of pomegranate, bilberry, and chokeberry at different concentrations were used as PSs. The unweighted (absolute) irradiance was 200 mW cm-2, applied five minutes. Planktonic cultures of Streptococcus mutans and Streptococcus sobrinus and total mixed bacteria from pooled saliva of volunteers were treated with aPDT. Up to more than 5 log10 of S. mutans and S. sobrinus were killed by aPDT with 0.4% and 0.8% pomegranate juice, 3% and 50% chokeberry juice, and 12.5% bilberry juice (both strains). Concentrations of at least 25% (pomegranate) and >50% (chokeberry and bilberry) eradicated the mixed bacteria in saliva samples. This pilot study has shown that pomegranate mother juice is superior to the berry juices as a multicomponent PS for killing pathogenic oral bacteria with aPDT.

Antimicrobial and Antibiofilm Effect of Hydrogel with Origanum vulgare on Culture of Streptococcus mutans and Streptococcus sobrinus / Efecto Antimicrobiano y Antibiofilm de Hidrogel a Base de Origanum vulgare sobre Cultivo de Streptococcus mutans y Streptococcus sobrinus

The oral cavity is an ecosystem that provides ideal conditions for the growth of bacteria, the Streptococcus genus is important for the formation of biofilms that lead to the development of dental caries, which affects the population worldwide. The world health organization encourages the use of plants thanks to its various therapeutic actions. Origanum vulgare L. (oregano), is an aromatic plant with medicinal and culinary properties. The objective of this study was to investigate the in vitro antimicrobial and antibiofilm activity of the ethanolic extract of oregano, against the growth of Streptococcus mutans and Streptococcus sobrinus ATCC. Leaves of the plant were obtained and the ethanolic extract was made by maceration. Antimicrobial activity was evaluated using the Kirby-Bauer method and compared with 2% chlorhexidine, subsequently the extract was incorporated into a hydrogel and its effect on biofilm formation was assessed by fluorescence microscopy and the main compounds were identified. present in the extratco. The study revealed that the extract presented antimicrobial effect against both strains and at 2% it showed high antimicrobial action compared to chlorhexidine at the same concentration, with average inhibition halos of 26.3 mm and 19 mm for each microorganism analyzed, (p < 0.05). Likewise, the hydrogel prepared with 2% extract significantly eliminated the preformed Streptococcus biofilm, at 24 hours of exposure, due to the presence of a variety of chemical groups, such as sterols, triterpenes, flavonoids, flavanones, flavanonol s, lactones. sesquiterpenic, tannins and coumarins. The oregano extract presented high antimicrobial action for both species, with a greater effect towards Streptococcus mutans and an interesting antibiofilm action; These results show the importance of exploring treatment alternatives of plant origin, to be considered as interesting complementary aids in dental therapy.

La cavidad oral es un ecosistema que proporciona condiciones ideales para el crecimiento de bacterias, el género Streptococcus es importante para la formación de biopelículas que conducen al desarrollo de caries dental, que afecta a la población a nivel mundial. La organización mundial de la salud, fomenta el uso de plantas gracias a sus diversas acciones terapéuticas. Origanum vulgare L. (orégano), es una planta aromática con propiedades medicinales y culinarias. El objetivo de este estudio fue investigar la actividad antimicrobiana y antibiofilm in vitro del extracto etanólico de oregano, contra el crecimiento de Streptococcus mutans y Streptococcus sobrinus ATCC. Se obtuvierón hojas de la planta y se realizó el extracto etanólico mediante maceración. La actividad antimicrobiana se evaluó mediante el método de Kirby-Bauer y se comparó con la clorhexidina al 2 %, posteriormente se incorporó el extracto en un hydrogel y se valoró su efecto sobre la formación del biofilm mediante microscopía de fluorescencia y se identificó los principales compuestos presentes en el extratco. El estudio reveló que el extracto presentó efecto antimicrobiano contra ambas cepas y al 2 % mostró alta acción antimicrobiana en comparación con la clorhexidina a la misma concentración, con halos de inhibición promedio de 26.3 mm y de 19 mm para cada microorganismo analizado, (p < 0.05). Así mismo, el hidrogel preparado con extracto al 2 %, eliminó significativamente la biopelícula preformada de Streptococcus, a las 24 horas de exposición, debido a la presencia de una variedad de grupos químicos, como esteroles, triterpenos, flavonoides, flavanonas, flavanonoles, lactonas sesquiterpénicas, taninos y cumarinas. El extracto de orégano presentó alta acción antimicrobiana para ambas especies, con mayor efecto hacia el Streptococcus mutans y una acción antibiofilm interesante; estos resultados muestran la importancia de explorar en alternativas de tratamiento de origen vegetal, para considerarse como auxiliares complementarios interesantes en la terapia dental.

Antimicrobial activity of ginger on cariogenic bacteria: molecular networking and molecular docking analyses.

Streptococcus mutans and Streptococcus sobrinus have been implicated as the primary causative agents of dental caries in humans. This study aimed to screen the antibacterial activity of the n-hexane, ethyl acetate, methanol, and aqueous extracts of Ginger against mentioned bacteria and investigate chemical constituents of the extracts, and their activity against some drug targets in S. mutans. Antimicrobial tests including biofilm inhibition, time-kill kinetics, and adherence inhibition alongside cytotoxicity of extracts, were assessed. A molecular networking technique was used to find chemical constituents of the extracts. Molecular docking analysis on the Schrodinger package was applied to identify the binding interactions of the compounds to targeted enzymes. Methanol and ethyl acetate extracts showed the highest antibacterial activity against S. mutans and S. sobrinus. Different compounds including polyphenols, alkaloids, anthraquinones, flavonoids, terpenoids, glycosides, steroids, and reducing sugars dereplicated from Ginger extracts. The binding affinity of ligands with free hydroxyl groups was better than other ligands against all tested enzymes. This study introduces a wide range of Z. officinal extracts compounds to be used in different drug discovery studies. Some Ginger compounds with high affinity to investigated enzymes can be considered as candidate compounds for anti-caries drug development studies.Communicated by Ramaswamy H. Sarma.

Inhibitory effects of citrus lemon oil and limonene on Streptococcus sobrinus - Induced dental caries in rats.

OBJECTIVE: This study aimed to examine and compare the anti-caries effects of citrus lemon oil (CLO) and limonene in rats. METHODS: The minimal inhibitory concentrations of CLO and limonene were measured using the disk diffusion method. The rats were infected with Streptococcus sobrinus and assigned into four groups: (1) Chlorhexidine, (2) CLO, (3) limonene, and (4) distilled water (H2O). The total cultivable microbiota and Streptococcus sobrinus in the mouth of the rats were counted, and the caries lesions were measured by Keyes' scoring and DIAGNOdent examination. RESULTS: The minimal inhibitory concentrations of CLO and limonene against Streptococcus sobrinus were 4.50 and 21.00 mg/mL, respectively. The chlorhexidine group had the lowest total microbiota counts (p < 0.05), while there were no significant differences among the CLO, limonene and H2O groups (p > 0.05). The proliferation of Streptococcus sobrinus was remarkably inhibited by chlorhexidine, limonene and CLO (p < 0.05). The Keyes' scoring and DIAGNOdent results indicated that the caries lesions were reduced in the CLO and limonene groups compared to that of the vehicle control group (p < 0.05). There was no significant difference between CLO and limonene (p > 0.05). CONCLUSION: Limonene and CLO have similar anti-caries abilities in a bacteriostatic manner in vivo.

Tea extracts differentially inhibit Streptococcus mutans and Streptococcus sobrinus biofilm colonization depending on the steeping temperature.

This study aimed to evaluate the effects of tea extracts on oral biofilm colonization depending on steeping temperature. S. mutans and S. sobrinus were cultured and treated with green or black tea extracts prepared under different steeping conditions. Biofilm formation, glucosyltransferase (GTF) levels, bacterial growth, and acidogenicity were evaluated. Biofilms were also assessed by gas chromatography-mass spectrometry and confocal laser scanning microscopy. All extracts with hot steeping showed higher inhibitory effects on biofilm formation and cell viability and lower GTF levels compared with those with cold steeping (p < 0.05). Hot steeping significantly reduced bacterial growth (p < 0.05) and maintained the pH. Catechins were only identified from hot steeping extracts. Within the limits of this study, extracts with cold steeping showed lower inhibitory effects on oral biofilms. The different effects between steeping extracts may be attributed to the difference in catechins released from tea extracts under the different steep conditions.

Inhibitory Effects of Lingonberry Extract on Oral Streptococcal Biofilm Formation and Bioactivity.

Phenolic compounds in fruits such as cranberries have been shown to promote a number of biological activities. The purpose of this study was to investigate the effects of polyphenolic compound-containing lingonberry extract on oral streptococci and compare them with the known anti-cariogenic activity of cranberries. Water-soluble and polyphenol-rich fractions (Fractions I and II, respectively) were isolated from cranberries and lingonberries. The effects of those fractions on the biofilm formation ability and bioactivity of Streptococcus mutans MT8148R, Streptococcus sobrinus 6715, and Streptococcus sanguinis ATCC 10556 were then evaluated. Cranberry or lingonberry Fraction II (at 0.5-1 mg/ml) significantly reduced biofilm formation by S. mutans, S. sobrinus, and S. sanguinis. In contrast, cranberry or lingonberry Fraction I (at 0.5-2 mg/ml) increased biofilm formation by S. mutans and S. sobrinus, but not by S. sanguinis. Fractions I and II (at 1-2 mg/ml) also reduced the bioactivity of S. mutans, while Fraction II (at 0.5 mg/ml) enhanced the bioactivity of all tested strains. The results revealed that lingonberries contained a larger amount of polyphenol than cranberries and that they showed almost the same level of activity against the biofilm formation ability and bioactivity of oral streptococci. This indicates that polyphenol-rich lingonberry fraction offers a promising natural food derivative for prevention of dental caries.

Physico-mechanical and antimicrobial properties of an orthodontic adhesive containing cationic curcumin doped zinc oxide nanoparticles subjected to photodynamic therapy.

BACKGROUND: Potential complications on the crown level during fixed orthodontic procedures are white spot, enamel demineralization and tooth decay. This study evaluated the antimicrobial properties of an orthodontic adhesive incorporating cationic curcumin doped zinc oxide nanoparticles (cCur/ZnONPs), which can have the highest concentration of cCur/ZnONPs and shear bond strength (SBS) value simultaneously, against cariogenic bacteria including Streptococcus mutans, Streptococcus sobrinus, and Lactobacillus acidophilus. MATERIALS AND METHODS: Following synthesis and confirmation of cCur/ZnONPs, SBS and adhesive remnant index (ARI) of the test adhesives containing cCur/ZnONPs (1.2, 2.5, 5, 7.5, and 10% wt.) were measured using universal testing machine and stereomicroscope, respectively. After continuously rinsed (up to 180 day), the residual antimicrobial ability of modified adhesives which can have the highest concentration of cCur/ZnONPs and SBS value simultaneously were determined by disc agar diffusion (DAD), biofilm formation inhibition, and metabolic activity assays following photo-activation using light-emitting diode (LED) for 5 min against multispecies cariogenic biofilm-producing bacteria. RESULTS: Adhesive with 7.5% wt. cCur/ZnONPs showed the highest concentration of cCur/ZnONPs and SBS value (14.89 ± 3.26 MPa, P < 0.05) simultaneously. No significant differences in ARI scores were found between the modified adhesive and control (Transbond XT without the cCur/ZnONPs). 7.5% wt. cCur/ZnONPs following photo-activation was not colonized by the test microorganisms and suppressed 100% metabolic activity of the test microorganisms up to 90 day compared to the control group (cCur/ZnONPs free LED irradiation; P < 0.05). In DAD assay, the reduction of photodynamic disinfection of the 7.5% wt. cCur/ZnONPs against test bacteria was positively associated to the time, in such a way that it was decreased significantly after 60 day. From days 120 onwards, microbial biofilm formation and metabolic activity was progressively increased on 7.5% wt. cCur/ZnONPs adhesive discs compared to the control group (cCur/ZnONPs free LED irradiation). CONCLUSIONS: Our findings highlight the photo-activated 7.5% wt. cCur/ZnONPs can serve as an orthodontic adhesive additive to control the cariogenic multispecies biofilm, and also to reduce their metabolic activity.

Antimicrobial and Antibiofilm Activities of Sulfated Polysaccharides from Marine Algae against Dental Plaque Bacteria.

Dental plaque biofilms cause various dental diseases; therefore, inhibiting the growths of the dental plaque bacteria which produce biofilms can be a strategy for preventing dental disease. Certain sulfated polysaccharides from marine algae exert antimicrobial activities against human bacterial pathogens in addition to their physiological benefits. On the basis of these observations, the antimicrobial and antibiofilm activities of sulfated polysaccharides from different marine algae were evaluated against dental plaque bacteria. Among the sulfated polysaccharides, a fucoidan from Fucus vesiculosus showed notable antimicrobial activities against the selected dental plaque bacteria, including some foodborne pathogenic bacteria. The minimum inhibitory concentrations were of 125 to 1000 µg mL-1. Regarding the antibiofilm activity, the fucoidan at the concentrations of above 250 µg mL-1 completely suppressed the biofilm formations and planktonic cell growths of Streptococcus mutans and S. sobrinus. However, no eliminative effect on the completed biofilm was observed. The fucoidan consisted of almost fucose base polysaccharide containing approximately 14.0% sulfate content. The average molecular weight of the fucoidan was changed by heat treatment (121 °C for 15 min) and it affected the antimicrobial activity.

Antimicrobial Effect of Polyphenon 60 against Streptococcus mutans and Streptococcus sobrinus

Polyphenon 60 refers to the mixture of catechins present in green tea. The aim of this study was to investigate the antimicrobial activities of polyphenon 60 against 4 strains of Streptococcus mutans and 2 strains of Streptococcus sorbrinus, which are the major causative bacteria of dental caries. The minimum bactericidal concentration (MBC) values of polyphenon 60 for S. mutans and S. sobrinus were determined and the effect of biofilm formation inhibition of that was evaluated. The MBC value of polyphenon 60 against the bacterial strains was 2.5 mg/ml except for one particular strain, S. mutans KCOM 1128 for which the value was 1.25 mg/ml. The results of biofilm formation inhibition assay revealed that polyphenon 60 inhibited biofilm formation more than 90% at a concentration of 2.5 mg/ml. It was apparent that polyphenon exhibited biofilm formation inhibition activity along with bactericidal effect against S. mutans and S. sobrinus. Therefore, it is proposed that polyphenon 60 as one of the components of bactericidal agents could be useful in developing oral hygiene products, toothpaste or gargling solution.

Galla chinensis Compounds Remineralize Enamel Caries Lesions in a Rat Model.

OBJECTIVES: To investigate the effect of Galla chinensis chemical compounds on enamel caries remineralization in rats. METHODS: A total of 40 rats were inoculated with Streptococcus sobrinus 6715 and fed a cariogenic diet (Keyes 2000). The rats were randomly divided into 4 groups and treated topically twice a day with each of the following extracts (or control) for 5 weeks: distilled and deionized water (DDW, negative control); 1,000 ppm NaF (positive control); 4,000 ppm G. chinensis crude aqueous extract (GCE), or 4,000 ppm gallic acid (GA). After the experimental period, Keyes' caries diagnosis and scoring technique was applied as a preliminary evaluation on the molar teeth. For more accurate remineralization data, the residual enamel volume of the first molar and the mineral density (MD) of the enamel were further analyzed by micro-CT. RESULTS: The severity of molar enamel caries decreased in the following order of treatment groups: DDW > GA > GCE > NaF (p < 0.05). In addition, the residual first molar enamel volume and MD values increased in the order of DDW, GA, GCE and NaF treatment groups (p < 0.05). In most molars, remineralization layers were observed on the surface of lesions. CONCLUSION: G. chinensis compounds remineralize enamel caries lesions in a rat model.

Antimicrobial effect of the extracts from Hypericum perforatum against oral bacteria and biofilm formation.

CONTEXT: One traditional medicines, Hypericum perforatum L. (Hypericaceae), possesses several beneficial effects against depression, ulcers, dyspepsia, abdominal pains, burns, bacterial infections, migraine headaches, and sciatica. OBJECTIVE: The present study investigates the antimicrobial activity of the ethanol extract (HP-EtOH) of H. perforatum and its sub-extracts, namely n-hexane (HP-hexane), chloroform (HP-CHCl3), ethyl acetate (HP-EtOAc), n-butanol (HP-n-BuOH), and water (HP-H2O) extracts, against Streptococcus mutans, S. sobrinus, Lactobacillus plantarum, and Enterococcus faecalis. MATERIALS AND METHODS: For the evaluation of the antimicrobial activity, flowering aerial parts of H. perforatum were extracted with EtOH and then this extract was fractionated to obtain five sub-extracts in different polarities. Antimicrobial activities of HP-EtOH and its sub-extracts against Streptococcus mutans, S. sobrinus, L. plantarum, and E. faecalis were assessed by using colorimetric micro-well dilution at concentration ranges of 64-0.5 µg/ml as well as resazurin microplate and modified microtiter-plate assays between the ranges of 100 and 0.78125 µg/ml. RESULTS: According to the results of the present study, HP-H2O sub-extract displayed strong antibacterial activity (MIC values 8 µg/mL) against S. sobrinus and L. plantarum, and exerted moderate activity against S. mutans and E. faecalis at 32 and 16 µg/mL concentrations, respectively. Other sub-extracts also demonstrated antimicrobial activity against S. sobrinus at a concentration of 16 µg/mL. HP-EtOAc and HP-n-BuOH showed antimicrobial activity against L. plantarum and HP-EtOAc and HP-H2O were also active against E. faecalis at the same concentrations (16 µg/mL). CONCLUSION: According to the results, we suggest that H. perforatum could be employed as a natural antibacterial agent in oral care products.

In vitro growth-inhibitory effect of ethanol GRAS plant and supercritical CO2 hop extracts on planktonic cultures of oral pathogenic microorganisms.

Conventional chemical antiseptics used for treatment of oral infections often produce side-effects, which restrict their long-term use. Plants are considered as perspective sources of novel natural antiseptics. However, little is still known about their inhibitory properties against oral pathogens. The objective of this study was to test in vitro antimicrobial activities of generally recognized as safe (GRAS) species against planktonic cultures of cariogenic, periodontal and candidal microorganisms and identify active compounds of the most active extracts. Growth-inhibitory effects of ethanol extracts from 109 GRAS plant species, six Humulus lupulus cultivars and two hop supercritical CO2 extracts were evaluated using broth microdilution method. The chemical analysis was done through high-performance liquid chromatography. Best results were obtained for supercritical CO2 and ethanol extracts of H. lupulus with minimum inhibitory concentrations (MIC) ≥8 µg/mL and ≥16 µg/mL, respectively. The chemical analysis of supercritical CO2H. lupulus extracts revealed that α- and ß-acids were their main constituents. Capsicum annuum and Capsicum frutescens showed antibacterial effect against Streptococcus sobrinus and Streptococcus salivarius (MIC=64-128 µg/mL). These strains were further inhibited by Zanthoxylum clava-herculis (MIC=64-128 µg/mL) and Myristica fragrans (both MIC≥128 µg/mL). The latter also exhibited antimicrobial activity against Fusobacterium nucleatum (MIC=64 µg/mL). Punica granatum possessed inhibitory effects against Candida albicans (MIC=128 µg/mL) and F. nucleatum (MIC=64 µg/mL). The results indicate that supercritical CO2H. lupulus extracts together with ethanol extracts of C. annuum, C. frutescens, M. fragrans, P. granatum and Z. clava-herculis are promising materials for further investigation on new antiseptic agents of oral care products.

In vitro antimicrobial activities of 1-methoxyficifolinol, licorisoflavan A, and 6,8-diprenylgenistein against Streptococcus mutans.

The objective of the study was to investigate the antimicrobial effects of purified single compounds from ethanol-extracted licorice root on Streptococcus mutans. The crude licorice root extract (CLE) was obtained from Glycyrrhiza uralensis, which was subjected to column chromatography to separate compounds. Purified compounds were identified by mass spectrometry and nuclear magnetic resonance. Antimicrobial activities of purified compounds from CLE were evaluated by determining the minimum inhibitory concentration and by performing time-kill kinetics. The inhibitory effects of the compounds on biofilm development were evaluated using crystal violet assay and confocal microscopy. Cell toxicity of substances to normal human gingival fibroblast (NHGF) cells was tested using a methyl thiazolyl tetrazolium assay. Chlorhexidine digluconate (CHX) was used in the control group. Three antimicrobial flavonoids, 1-methoxyficifolinol, licorisoflavan A, and 6,8-diprenylgenistein, were isolated from the CLE. We found that the three flavonoids and CHX had bactericidal effects on S. mutans UA159 at the concentration of ≥4 and ≥1 µg/ml, respectively. The purified compounds completely inhibited biofilm development of S. mutans UA159 at concentrations over 4 µg/ml, which was equivalent to 2 µg/ml of CHX. Confocal analysis showed that biofilms were sparsely scattered in the presence of over 4 µg/ml of the purified compounds. However, the three compounds purified from CLE showed less cytotoxic effects on NHGF cells than CHX at these biofilm-inhibitory concentrations. Our results suggest that purified flavonoids from CLE can be useful in developing oral hygiene products, such as gargling solutions and dentifrices for preventing dental caries.

Rapid and sensitive PCR-dipstick DNA chromatography for multiplex analysis of the oral microbiota.

A complex of species has been associated with dental caries under the ecological hypothesis. This study aimed to develop a rapid, sensitive PCR-dipstick DNA chromatography assay that could be read by eye for multiplex and semiquantitative analysis of plaque bacteria. Parallel oligonucleotides were immobilized on a dipstick strip for multiplex analysis of target DNA sequences of the caries-associated bacteria, Streptococcus mutans, Streptococcus sobrinus, Scardovia wiggsiae, Actinomyces species, and Veillonella parvula. Streptavidin-coated blue-colored latex microspheres were to generate signal. Target DNA amplicons with an oligonucleotide-tagged terminus and a biotinylated terminus were coupled with latex beads through a streptavidin-biotin interaction and then hybridized with complementary oligonucleotides on the strip. The accumulation of captured latex beads on the test and control lines produced blue bands, enabling visual detection with the naked eye. The PCR-dipstick DNA chromatography detected quantities as low as 100 pg of DNA amplicons and demonstrated 10- to 1000-fold higher sensitivity than PCR-agarose gel electrophoresis, depending on the target bacterial species. Semiquantification of bacteria was performed by obtaining a series of chromatograms using serial 10-fold dilution of PCR-amplified DNA extracted from dental plaque samples. The assay time was less than 3 h. The semiquantification procedure revealed the relative amounts of each test species in dental plaque samples, indicating that this disposable device has great potential in analysis of microbial composition in the oral cavity and intestinal tract, as well as in point-of-care diagnosis of microbiota-associated diseases.

Systematic screening of plant extracts from the Brazilian Pantanal with antimicrobial activity against bacteria with cariogenic relevance.

This study proposes a bioprospection methodology regarding the antimicrobial potential of plant extracts against bacteria with cariogenic relevance. Sixty extracts were obtained from ten plants--(1) Jatropha weddelliana, (2) Attalea phalerata, (3) Buchenavia tomentosa, (4) Croton doctoris, (5) Mouriri elliptica, (6) Mascagnia benthamiana, (7) Senna aculeata, (8) Unonopsis guatterioides, (9) Allagoptera leucocalyx and (10) Bactris glaucescens--using different extraction methods - (A) 70° ethanol 72 h/25°C, (B) water 5 min/100°C, (C) water 1 h/55°C, (D) water 72 h/25°C, (E) hexane 72 h/25°C and (F) 90° ethanol 72 h/25°C. The plants were screened for antibacterial activity at 50 mg/ml using the agar well diffusion test against Actinomyces naeslundii ATCC 19039, Lactobacillus acidophilus ATCC 4356, Streptococcus gordonii ATCC 10558, Streptococcus mutans ATCC 35688, Streptococcus sanguinis ATCC 10556, Streptococcus sobrinus ATCC 33478 and Streptococcus mitis ATCC 9811. The active extracts were tested to determine their minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), cytotoxicity and chemical characterization. Forty-seven extracts (78%) were active against at least one microorganism. Extract 4A demonstrated the lowest MIC and MBC for all microorganisms except S. gordonii and the extract at MIC concentration was non-cytotoxic. The concentrated extracts were slightly cytotoxic. Electrospray ionization with tandem mass spectrometry analyses demonstrated that the extract constituents coincided with the mass of the terpenoids and phenolics. Overall, the best results were obtained for extraction methods A, B and C. The present work proved the antimicrobial activity of several plants. Particularly, extracts from C. doctoris were the most active against bacteria involved in dental caries disease.

Effects of adjunctive daily phototherapy on chronic periodontitis: a randomized single-blind controlled trial

PURPOSE: The purpose of this randomized single-blind controlled trial was to elucidate the clinical and antimicrobial effects of daily phototherapy (PT) as an adjunct to scaling and root planing (SRP) in patients with chronic periodontitis. METHODS: The study was conducted from December 2013 to May 2014 at Ewha Womans University Mokdong Hospital, Seoul, Korea. Forty-one patients with mild to moderate chronic periodontitis were randomly divided into two therapeutic groups in a 1:1 ratio: SRP+PT and SRP (control) groups. All participants underwent full-mouth SRP. PT was performed thrice a day for a month by using electric toothbrushes with embedded light-emitting diodes. Plaque index, gingival index, probing pocket depth (PPD), clinical attachment level (CAL), and bleeding on probing were assessed before (baseline) and four weeks after (follow-up) the treatment. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum, Parvimonas micra, Campylobacter rectus, Eikenella corrodens, Streptococcus mutans, and Streptococcus sobrinus levels were detected by a real-time polymerase chain reaction at the same points in time. RESULTS: The clinical parameters improved in both the groups. At the follow-up assessment, PPD was significantly decreased in the SRP+PT group (P=0.00). Further, PPD and CAL showed significantly greater changes in the SRP+PT group than in the SRP group (PPD, P=0.03; CAL, P=0.04). P. gingivalis and T. forsythia levels decreased in this group, but no significant intergroup differences were noted. CONCLUSIONS: Adjunctive PT seems to have clinical benefits, but evidence of its antimicrobial effects is not sufficient. Long-term studies are necessary to develop the most effective PT protocol and compare the effectiveness of PT with and without exogenous photosensitizers.

Antimicrobial activity of fractions and subfractions of Elaeagia utilis against microorganisms of importance in dental caries.

Dental caries is a multifactorial infectious disease that leads to the destruction of dental hard tissue. The main goal of research into medicinal plants is to seek compounds with antimicrobial activity for subsequent use in prevention strategies and control of infectious diseases. The aim of this study was to evaluate the antimicrobial activity of fractions and subfractions obtained from Elaeagia utilis against Streptococcus mutans, Streptococcus sobrinus and Lactobacillus acidophilus. The plant material was collected in the town of Alban (Cundinamarca, Colombia), which is located at an altitude of 2245 meters above sea level. Two extracts were obtained by cold maceration of E. utilis leaves in (a) petroleum ether extract and (b) ethanol extract. Fractions were obtained from the petroleum ether extract by column vacuum chromatography, and from the ethanol extract by continuous liquid/liquid partitioning. The antimicrobial activity of fractions and subfractions was evaluated by the well diffusion method. At a concentration of 10 mg/well, several fractions from both extracts showed antimicrobial activity against S. mutans, S. sobrinus and L. acidophilus. Among the ethanol extract fractions, the dichloromethane fraction had notably greater antimicrobial activity. It was sub-partitioned, yielding three subfractions with inhibitory activity, of which the most active was MeOH: H2O (Bp) with minimum inhibitory concentration 0.1 mg/well on the 3 study bacteria. Terpenes, sesquiterpenlactones and simple phenolic compounds were identified in it. In conclusion, this study shows the antimicrobial potential of fractions and subfractions obtained from extracts of E. utilis leaves against bacteria that are important in dental caries.

Chemical and botanical characterization of Chilean propolis and biological activity on cariogenic bacteria Streptococcus mutans and Streptococcus sobrinus.

Propolis is a non-toxic natural substance with multiple pharmacological properties including anti-cancer, antioxidant, fungicidal, antibacterial, antiviral, and anti-inflammatory among others. The aim of this study was to determine the chemical and botanical characterization of Chilean propolis samples and to evaluate their biological activity against the cariogenic bacteria Streptococcus mutans and Streptococcus sobrinus. Twenty propolis samples were obtained from beekeeping producers from the central and southern regions of Chile. The botanical profile was determined by palynological analysis. Total phenolic contents were determined using colorimetric assays. Reverse phase HPLC and HPLC-MS were used to determine the chemical composition. The minimum inhibitory concentration (MIC) was determined on S. mutans and S. sobrinus. All propolis samples were dominated by structures from native plant species. The characterization by HPLC/MS, evidenced the presence of quercetin, myricetin, kaempferol, rutine, pinocembrin, coumaric acid, caffeic acid and caffeic acid phenethyl ester, that have already been described in these propolis with conventional HPLC. Although all propolis samples inhibited the mutans streptococci growth, it was observed a wide spectrum of action (MIC 0.90 to 8.22 µg mL(-1)). Given that results it becomes increasingly evident the need of standardization procedures, where we combine both the determination of botanical and the chemical characterization of the extracts. Research conducted to date, describes a promising effectiveness of propolis in the prevention of caries and other diseases of the oral cavity, making it necessary to develop studies to identify and understand the therapeutic targets or mechanisms of molecular action of the various compounds present on them.

Adhesion and biofilm formation by oral streptococci on different commercial brackets.

AIM: To compare early bacterial adhesion and biofilm formation in vitro by different oral streptococci on a variety of commercial brackets. METHODS: Adhesion and biofilm formation in vitro of 6 Streptococcus spp. on 15 different commercial brackets, in standard culture medium and in human saliva were evaluated by the MTT reduction assay. RESULTS: Significant differences were evidenced in both early adhesion and biofilm formation among the studied brackets and between the two conditions of growth. Gold brackets resulted less prone to colonisation compared to composite brackets. The growth rates of the tested species on the different tested materials were significantly different. CONCLUSION: The adopted experimental plan, dissecting the two phases of plaque formation on different brackets in different conditions, showed that composite brackets are more susceptible to adhesion and colonisation by streptococci, while the remaining tested brackets did not show differences that could be clinically relevant. Data suggest that different personal behaviours affecting the oral environment could significantly affect colonisation of brackets by oral streptococci.