RESUMEN
Streptococcus gordonii is a commensal colonizer of oral cavity that initiates the formation of dental plaque. Oxyresveratrol is a natural purification from plants with antibacterial effects on various oral bacteria including Streptococcus mutans. The aim of this study was to investigate the effects of oxyresveratrol on S. gordonii. The basic viability, biofilm formation and cell aggregation of S. gordonii treated with oxyresveratrol were investigated. Oxyresveratrol dose-dependently inhibited the growth of S. gordonii in the absence of sucrose. However, in the presence of sucrose, it promoted biofilm formation under MIC. Both the biofilm formation and extracellular polysaccharides synthesis reached the maximum level at ½ MIC (250 µg/mL) oxyresveratrol. The gene expressions of abpA, abpB, scaA, gtfG, hsa, cshA, cshB, ccpA, srtA and sspB were upregulated when treated with 62.5 and 125 µg/mL oxyresveratrol. A total eight of the ten genes were significantly upregulated at 250 µg/mL oxyresveratrol except abpB and sspB, which were downregulated at 250 µg/mL without significance. In conclusion, oxyresveratrol has dual-effects on S. gordonii. Considering its specific biofilm suppressive effect on S. mutans, it might be a candidate for bacterial interspecies modulator applied in caries prevention.
Asunto(s)
Adhesión Bacteriana/efectos de los fármacos , Biopelículas/efectos de los fármacos , Extractos Vegetales/farmacología , Estilbenos/farmacología , Streptococcus gordonii/efectos de los fármacos , Sacarosa/farmacología , Antiinfecciosos/farmacología , Interacciones Farmacológicas , Perfilación de la Expresión Génica , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Streptococcus gordonii/genéticaRESUMEN
The pyruvate oxidase (SpxB)-dependent production of H2O2 is widely distributed among oral commensal streptococci. Several studies confirmed the ability of H2O2 to antagonize susceptible oral bacterial species, including caries-associated Streptococcus mutans as well as several periodontal pathobionts. Here we report a potential mechanism to bolster oral commensal streptococcal H2O2 production by magnesium (Mg2+) supplementation. Magnesium is a cofactor for SpxB catalytic activity, and supplementation increases the production of H2O2 in vitro. We demonstrate that Mg2+ affects spxB transcription and SpxB abundance in Streptococcus sanguinis and Streptococcus gordonii. The competitiveness of low-passage commensal streptococcal clinical isolates is positively influenced in antagonism assays against S. mutans. In growth conditions normally selective for S. mutans, Mg2+ supplementation is able to increase the abundance of S. sanguinis in dual-species biofilms. Using an in vivo biophotonic imaging platform, we further demonstrate that dietary Mg2+ supplementation significantly improves S. gordonii oral colonization in mice. In summary, our results support a role for Mg2+ supplementation as a potential prebiotic to promote establishment of oral health-associated commensal streptococci.
Asunto(s)
Boca , Animales , Biopelículas , Peróxido de Hidrógeno , Magnesio , Ratones , Streptococcus gordonii , Streptococcus mutans , Streptococcus sanguisRESUMEN
We have previously demonstrated that out of the butyrolactones series synthesized based on the natural lichen metabolite lichesterinic acid, compound (B-13) was the most effective against oral bacteria. However, its antibacterial mechanism is still unknown. In this study, we have investigated its bacterial localization by synthesizing a fluorescently labeled B-13 with NBD while maintaining its antibacterial activity. We showed that this compound binds to Streptococcus gordonii cell surface, as demonstrated by HPLC analysis. By adhering to cell surface, B-13 induced cell wall disruption leading to the release of bacterial constituents and consequently, the death of S. gordonii, a Gram-positive bacterium. A Gram-negative counterpart, Porphyromanas gingivalis, showed also cracked and ruptured cells in the presence of B-13. Besides, we also demonstrated that the analog of B-13, B-12, has also induced disruption of P. gingivalis and S. gordonii. This study revealed that butyrolactones can be considered as potent antibacterial compounds against oral pathogens causing medical complications.
Asunto(s)
Antibacterianos/farmacología , Lactonas/farmacología , Líquenes/química , Porphyromonas gingivalis/efectos de los fármacos , Streptococcus gordonii/efectos de los fármacos , Membrana Celular/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Estructura MolecularRESUMEN
Objective: The purpose of this in vitro study was to evaluate the antimicrobial effect of activated irrigation with different modes of erbium-doped yttrium aluminum garnet (Er:YAG) laser application on microorganisms related to secondary endodontic infection. Background: Er:YAG laser has been recommended as an adjuvant tool for root canal disinfection during endodontic treatment. Materials and methods: Laser-activated irrigation (LAI) with 300 or 600 µm tips were tested with or without intermittent irrigation with 0.9% sodium chloride (NaCl) solution against different microorganisms (five single strains and dual species (Streptococcus gordonii combined with Actinomyces oris or Fusobacterium nucleatum) in root canals after 3 days of incubation. In a 21-day infection model, LAI was used together with intermittent rinsing with sodium hypochlorite (NaOCl) against the dual-species mixtures; here the incidence of microbial regrowth after up to 7 days was monitored. Results: In the 3-day root infection model, LAI protocols did not show any significant reduction of the microbial load when compared with manual irrigation with saline solution. In the 21-day infection, S. gordonii combined with A. oris were not detectable anymore after applying the LAI protocol with a 600 µm tip (30 mJ/10 pps) up to 7 days after treatment. Conclusions: Application of LAI with a 600 µm tip by using an Er:YAG laser might be advantageous in treatment of endodontic infections.
Asunto(s)
Cavidad Pulpar/microbiología , Desinfección/instrumentación , Láseres de Estado Sólido , Preparación del Conducto Radicular/métodos , Irrigación Terapéutica/instrumentación , Actinomyces/efectos de la radiación , Candida albicans/efectos de la radiación , Enterococcus faecalis/efectos de la radiación , Fusobacterium nucleatum/efectos de la radiación , Técnicas In Vitro , Irrigantes del Conducto Radicular/farmacología , Hipoclorito de Sodio/farmacología , Streptococcus gordonii/efectos de la radiaciónRESUMEN
A novel silver nanoparticle (AgNP) formulation was developed as a targeted application for the disinfection of carious dentine. Silver nitrate (AgNO3) was chemically reduced using sodium borohydrate (NaBH4) in the presence of sodium dodecyl sulfate (SDS) to form micelle aggregate structures containing monodisperse 6.7- to 9.2-nm stabilized AgNPs. AgNPs were characterized by measurement of electrical conductivity and dynamic light scattering, scanning electron microscopy, transmission electron microscopy, and inductively coupled plasma mass spectrometry. Antimicrobial activity of AgNPs was tested against planktonic cultures of representative gram-positive and gram-negative oral bacteria using well diffusion assays on tryptic soy broth media and monoculture biofilms grown with brain heart infusion ± sucrose anaerobically at 37°C on microtiter plates. Biofilm mass was measured by crystal violet assay. Effects were compared to silver diamine fluoride and chlorhexidine (negative controls) and 70% isopropanol (positive control) exposed cultures. In the presence of AgNPs, triplicate testing against Streptococcus gordonii DL1, C219, G102, and ATCC10558 strains; Streptococcus mutans UA159; Streptococcus mitis I18; and Enterococcus faecalis JH22 for planktonic bacteria, the minimum inhibitory concentrations were as low as 7.6 µg mL-1 and the minimum bacteriocidal concentrations as low as 19.2 µg mL-1 silver concentration. Microplate readings detecting crystal violet light absorption at 590 nm showed statistically significant differences between AgNP-exposed biofilms and where no antimicrobial agents were used. The presence of sucrose did not influence the sensitivity of any of the bacteria. By preventing in vitro biofilm formation for several Streptococcus spp. and E. faecalis, this AgNP formulation demonstrates potential for clinical application inhibiting biofilms.
Asunto(s)
Biopelículas/efectos de los fármacos , Desinfectantes/farmacología , Nitrato de Plata/farmacología , Clorhexidina/farmacología , Caries Dental/microbiología , Desinfectantes/química , Conductividad Eléctrica , Enterococcus faecalis/efectos de los fármacos , Fluoruros Tópicos/farmacología , Nanopartículas del Metal/química , Pruebas de Sensibilidad Microbiana , Microscopía Electrónica , Compuestos de Amonio Cuaternario/farmacología , Compuestos de Plata/farmacología , Espectrofotometría Atómica , Streptococcus gordonii/efectos de los fármacos , Streptococcus mitis/efectos de los fármacos , Streptococcus mutans/efectos de los fármacosRESUMEN
The oral bacteria not only infect the mouth and reside there, but also travel through the blood and reach distant body organs. If left untreated, the dental biofilm that can cause destructive inflammation in the oral cavity may result in serious medical complications. In dental biofilm, Streptococcus gordonii, a primary oral colonizer, constitutes the platform on which late pathogenic colonizers like Porphyromonas gingivalis, the causative agent of periodontal diseases, will bind. The aim of this study was to determine the antibacterial activity of eleven natural lichen compounds belonging to different chemical families and spanning from linear into cyclic and aromatic structures to uncover new antibiotics which can fight against the oral bacteria. The compounds were screened by broth microdilution assay. Three compounds were shown to have promising antibacterial activities where the depsidone core with certain functional groups constituted the best compound, psoromic acid, with the lowest MICs=11.72 and 5.86µg/mL against S. gordonii and P. gingivalis, respectively. The compounds screened had promising antibacterial activity which might be attributed to some important functional groups as discussed in our study. The best compounds did not induce the death of gingival epithelial carcinoma cells (Ca9-22). These results introduce new compounds having potent antibacterial activities against oral pathogens causing serious medical complications.
Asunto(s)
Antibacterianos/química , Líquenes/química , Porphyromonas gingivalis/efectos de los fármacos , Streptococcus gordonii/efectos de los fármacos , Antibacterianos/aislamiento & purificación , Biopelículas/efectos de los fármacos , Línea Celular Tumoral , Humanos , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Boca/microbiología , Enfermedades Periodontales/microbiologíaRESUMEN
BACKGROUND AND OBJECTIVE: In the therapy of destructive periodontal disease, chemical antimicrobial agents and increasingly photodynamic therapy (PDT) play an important adjunctive role to standard mechanical anti-infective treatment procedures. However, both antiseptic methods have their shortcomings in terms of eliminating periodontal pathogens. The aim of the study was to compare the antibacterial efficacy of different antiseptic mouthrinses, of a conventional and a new, modified PDTplus as well as of the different antiseptic mouthrinses combined with either the conventional or the modified PDTplus against periopathogens. MATERIAL AND METHODS: Six representative periodontitis-associated bacterial strains were grown for 24 h under anaerobic conditions. After mixing the individual cell pellets they were exposed to 10 different antiseptic mouthrinse formulations: chlorhexidine (0.2%, 0.06%, CHX); CHX + cetylpyridinium chloride (each 0.05%); sodium hypochlorite (0.05%); polyhexanide (0.04%, PHMB1; 0.1%, PHMB2); octenidine dihydrochloride (0.1%); fluoride (250 ppm); essential oils; povidone iodine (10%); and saline (0.9%, NaCl) as control. Furthermore, the bacteria were treated with conventional PDT based on light-emitting diodes and a new modified photodisinfection combining photosensitizer with hydrogen peroxide to PDTplus also based on light-emitting diodes. In addition to the single treatments, a combined application of antiseptic exposure followed by use of PDT or PDTplus was performed. The microbial viability was characterized by analyzing colony growth and fluorescence-based vitality proportions. RESULTS: Nearly all mouthrinses caused a statistically significant growth inhibition. The most effective antiseptics, CHX (0.2%), CHX/cetylpyridinium chloride and octenidine dihydrochloride, inhibited bacterial growth completely. Conventional PDT resulted in moderate reduction of colony growth. The modified PDTplus achieved maximum antimicrobial effect. The combination of antiseptic exposure and PDT against periopathogens predominantly increased antibacterial efficacy compared to the single applications. The mouthrinse containing essential oil seemed to interfere with PDT. CONCLUSION: A combination therapy of preceding chemotherapeutical exposure and subsequent photodisinfection may be a more effective and promising antibacterial treatment than single applications of the antiseptic methods. The modified PDTplus using oxygen-enriched toluidine showed a superior antibacterial effect on periodontal pathogens to conventional PDT and to the majority of the investigated mouthrinses.
Asunto(s)
Actinomyces viscosus/efectos de los fármacos , Aggregatibacter actinomycetemcomitans/efectos de los fármacos , Antiinfecciosos Locales/farmacología , Fusobacterium nucleatum/efectos de los fármacos , Antisépticos Bucales/farmacología , Fotoquimioterapia , Porphyromonas gingivalis/efectos de los fármacos , Streptococcus gordonii/efectos de los fármacos , Veillonella/efectos de los fármacos , Terapia Combinada , Fotoquimioterapia/métodosRESUMEN
In the present study, antimicrobial activity of Piper betle crude ethanol extract against 4 strains of oral pathogens; Candida albicans DMST 8684, C. albicans DMST 5815, Streptococcus gordonii DMST 38731 and Streptococcus mutans DMST 18777 was compared with other medicinal plants. P. betle showed the strongest antimicrobial activity against all tested strains. Fractionated extracts of P. betle using hexane, ethyl acetate, and ethanol, respectively, were subjected to antimicrobial assay. The result revealed that the fractionated extract from ethyl acetate (F-EtOAc) possessed the strongest antimicrobial activity against all tested strains. Its inhibition zones against those pathogens were 23.00 ± 0.00, 24.33 ± 0.58, 12.50 ± 0.70 and 11.00 ± 0.00 mm, respectively and its minimum inhibitory concentrations were 0.50, 1.00, 0.50 and 1.00 mg/mL, respectively. Interestingly, the minimum concentration to completely kill those pathogens was the same for all strains and found to be 2.00 mg/mL. Killing kinetic study revealed that the activity of F-EtOAc was dose dependent. HPLC chromatograms of P. betle extracts were compared with its antimicrobial activity. An obvious peak at a retention time of 4.11 min was found to be a major component of F-EtOAc whereas it was a minor compound in the other extracts. This peak was considered to be an active compound of P. betle as it was consistent with the antimicrobial activity of F-EtOAc, the most potential extract against the tested pathogens. It is suggested that F-EtOAc is a promising extract of P. betle for inhibition of oral pathogens. Separation and structure elucidation of the active compound of this extract will be further investigated.
Asunto(s)
Antiinfecciosos/farmacología , Candida albicans/efectos de los fármacos , Piper betle , Extractos Vegetales/farmacología , Streptococcus gordonii/efectos de los fármacos , Streptococcus mutans/efectos de los fármacos , Acetatos , Cromatografía Líquida de Alta Presión , Etanol , Hexanos , Humanos , Pruebas de Sensibilidad Microbiana , Boca/microbiología , SolventesRESUMEN
Objective The clinical part of this study aimed to investigate whether the racemate of delmopinol [(±)-delmopinol] is equivalent to its two enantiomers [(+)-delmopinol and (-)-delmopinol] with respect to efficiency and to determine and compare their pharmacokinetic properties. The purpose of the pre-clinical part was to elucidate possible differences in antimicrobial efficiency. Materials and methods The compounds were tested clinically in a double-blind, randomized, cross-over study comprising three treatment periods of 4 days each. The antimicrobial efficacy of the enantiomers was compared in vitro with respect to planktonic and biofilm bacteria of different species. Results No statistically significant differences in prevention of plaque formation were observed. Except for a somewhat higher systemic exposure in terms of AUC and Cmax indicated for (-)-delmopinol compared to (+)-delmopinol, the pharmacokinetic properties were similar. The most common adverse event was a transient anaesthetic feeling in the mouth. This event was reported with the same frequency for all three test solutions. The enantiomers showed similar antimicrobial effects on planktonic bacteria and their biofilms. Conclusions The enantiomers were found to be equally effective with respect to inhibition of plaque development and only minor differences were observed with respect to their pharmacokinetic properties. No differences could be observed in the adverse events reports. There is, therefore, no reason to use one of the enantiomers of delmopinol instead of the racemate. This was further supported by the antimicrobial tests. It is suggested that the combined action of cationic and neutral delmopinol is important for its effect on biofilms.
Asunto(s)
Antibacterianos/uso terapéutico , Morfolinas/uso terapéutico , Antisépticos Bucales/uso terapéutico , Adolescente , Adulto , Antibacterianos/sangre , Antibacterianos/farmacocinética , Biopelículas/efectos de los fármacos , Estudios Cruzados , Placa Dental/microbiología , Placa Dental/prevención & control , Método Doble Ciego , Escherichia coli/efectos de los fármacos , Femenino , Gingivitis/prevención & control , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Morfolinas/sangre , Morfolinas/farmacocinética , Antisépticos Bucales/farmacocinética , Neisseria sicca/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacos , Staphylococcus epidermidis/efectos de los fármacos , Estereoisomerismo , Streptococcus gordonii/efectos de los fármacos , Adulto JovenRESUMEN
Arginine is utilized by the oral inhabitant Streptococcus gordonii as a substrate of the arginine deiminase system (ADS), eventually producing ATP and NH3, the latter of which is responsible for microbial resistance to pH stress. S. gordonii expresses a putative arginine-ornithine antiporter (ArcD) whose function has not been investigated despite relevance to the ADS and potential influence on inter-bacterial communication with periodontal pathogens that utilize amino acids as a main energy source. Here, we generated an S. gordonii ΔarcD mutant to explore the role of ArcD in physiological homeostasis and bacterial cross-feeding. First, we confirmed that S. gordonii ArcD plays crucial roles for mediating arginine uptake and promoting bacterial growth, particularly under arginine-limited conditions. Next, metabolomic profiling and transcriptional analysis of the ΔarcD mutant revealed that deletion of this gene caused intracellular accumulation of ornithine leading to malfunction of the ADS and suppression of de novo arginine biosynthesis. The mutant strain also showed increased susceptibility to low pH stress due to reduced production of ammonia. Finally, accumulation of Fusobacterium nucleatum was found to be significantly decreased in biofilm formed by the ΔarcD mutant as compared with the wild-type strain, although ornithine supplementation restored fusobacterium biovolume in dual-species biofilms with the ΔarcD mutant and also enhanced single species biofilm development by F. nucleatum. Our results are the first direct evidence showing that S. gordonii ArcD modulates not only alkali and energy production but also interspecies interaction with F. nucleatum, thus initiating a middle stage of periodontopathic biofilm formation, by metabolic cross-feeding.
Asunto(s)
Sistemas de Transporte de Aminoácidos/metabolismo , Antiportadores/metabolismo , Arginina/metabolismo , Proteínas Bacterianas/metabolismo , Biopelículas , Infecciones Estreptocócicas/microbiología , Streptococcus gordonii/fisiología , Sistemas de Transporte de Aminoácidos/genética , Antiportadores/genética , Proteínas Bacterianas/genética , Biopelículas/crecimiento & desarrollo , Fusobacterium nucleatum/fisiología , Eliminación de Gen , Humanos , Interacciones Microbianas , Ornitina/metabolismo , Streptococcus gordonii/genética , Streptococcus gordonii/crecimiento & desarrolloRESUMEN
OBJECTIVE: The aim of the present study was to evaluate in vitro antibacterial activities of blackcurrant and sea buckthorn juices on bacteria associated with gingival inflammation. MATERIALS AND METHODS: The growth of selected bacteria (Streptococcus mitis, Streptococcus mutans, Streptococcus sanguinis, Streptococcus gordonii, Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa) was studied in vitro on agar plates. The content of phenols in the different extracts was measured with HPLC-ESI-MS. RESULTS: The spectrometric analysis identified that the highest level of the single phenols studied was found for ferulic acid (113 µg/ml) in blackcurrant juice. Sea buckthorn contained low levels of selected phenols. Total bacterial inhibition for all bacterial species studied was found at 20% berry juice concentration with pH varying between 4.1-5.4. CONCLUSIONS: The present study identified that in vitro bacterial growth on agar plates was inhibited by blackcurrant and sea buckthorn juices and that low juice pH explains bacterial in vitro growth. This may have clinical implications in biofilm development, reducing the risks for both tooth decay and gingivitis.
Asunto(s)
Antibacterianos/farmacología , Frutas , Hippophae , Extractos Vegetales/farmacología , Ribes , Técnicas Bacteriológicas , Biopelículas/efectos de los fármacos , Ácido Clorogénico/análisis , Cromatografía Líquida de Alta Presión , Ácidos Cumáricos/análisis , Humanos , Concentración de Iones de Hidrógeno , Ensayo de Materiales , Fenoles/análisis , Fenoles/farmacología , Extractos Vegetales/análisis , Pseudomonas aeruginosa/efectos de los fármacos , Espectrometría de Masa por Ionización de Electrospray , Staphylococcus aureus/efectos de los fármacos , Staphylococcus epidermidis/efectos de los fármacos , Streptococcus gordonii/efectos de los fármacos , Streptococcus mutans/efectos de los fármacos , Streptococcus sanguis/efectos de los fármacosRESUMEN
The purpose of this study was to evaluate various biological effects of silver-zeolite and a polyphenol-rich extract of A. nodosum (ASCOP) to prevent and/or treat biofilm-related oral diseases. Porphyromonas gingivalis and Streptococcus gordonii contribute to the biofilm formation associated with chronic periodontitis. In this study, we evaluated in vitro antibacterial and anti-biofilm effects of silver-zeolite (Ag-zeolite) combined to ASCOP on P. gingivalis and S. gordonii growth and biofilm formation capacity. We also studied the anti-inflammatory and antioxidant capacities of ASCOP in cell culture models. While Ag-zeolite combined with ASCOP was ineffective against the growth of S. gordonii, it showed a strong bactericidal effect on P. gingivalis growth. Ag-zeolite combined with ASCOP was able to completely inhibit S. gordonii monospecies biofilm formation as well as to reduce the formation of a bi-species S. gordonii/P. gingivalis biofilm. ASCOP alone was ineffective towards the growth and/or biofilm formation of S. gordonii and P. gingivalis while it significantly reduced the secretion of inflammatory cytokines (TNFα and IL-6) by LPS-stimulated human like-macrophages. It also exhibited antioxidant properties and decreased LPS induced lipid peroxidation in gingival epithelial cells. These findings support promising use of these products in future preventive or therapeutic strategies against periodontal diseases.
Asunto(s)
Ascophyllum/química , Extractos Vegetales/farmacología , Polifenoles/farmacología , Plata/farmacología , Zeolitas/farmacología , Antibacterianos/farmacología , Antiinflamatorios/farmacología , Antioxidantes/farmacología , Biopelículas/efectos de los fármacos , Línea Celular Tumoral , Citocinas/metabolismo , Humanos , Enfermedades Periodontales/microbiología , Enfermedades Periodontales/prevención & control , Porphyromonas gingivalis/efectos de los fármacos , Plata/química , Streptococcus gordonii/efectos de los fármacos , Zeolitas/químicaRESUMEN
The oral pathogen Aggregatibacter actinomycetemcomitans (Aa) resides in infection sites with many microbes, including commensal streptococci such as Streptococcus gordonii (Sg). During infection, Sg promotes the virulence of Aa by producing its preferred carbon source, l-lactate, a phenomenon referred to as cross-feeding. However, as with many streptococci, Sg also produces high levels of the antimicrobial hydrogen peroxide (H2O2), leading to the question of how Aa deals with this potent antimicrobial during coinfection. Here, we show that Aa possesses two complementary responses to H2O2: a detoxification or fight response mediated by catalase (KatA) and a dispersion or flight response mediated by Dispersin B (DspB), an enzyme that dissolves Aa biofilms. Using a murine abscess infection model, we show that both of these responses are required for Sg to promote Aa virulence. Although the role of KatA is to detoxify H2O2 during coinfection, 3D spatial analysis of mixed infections revealed that DspB is required for Aa to spatially organize itself at an optimal distance (>4 µm) from Sg, which we propose allows cross-feeding but reduces exposure to inhibitory levels of H2O2. In addition, these behaviors benefit not only Aa but also Sg, suggesting that fight and flight stimulate the fitness of the community. These results reveal that an antimicrobial produced by a human commensal bacterium enhances the virulence of a pathogenic bacterium by modulating its spatial location in the infection site.
Asunto(s)
Aggregatibacter actinomycetemcomitans/patogenicidad , Proteínas Bacterianas/metabolismo , Biopelículas/crecimiento & desarrollo , Catalasa/metabolismo , Coinfección/fisiopatología , Glicósido Hidrolasas/metabolismo , Streptococcus gordonii/metabolismo , Aggregatibacter actinomycetemcomitans/metabolismo , Animales , Coinfección/microbiología , Peróxido de Hidrógeno/metabolismo , Ácido Láctico/metabolismo , Ratones , Análisis por Micromatrices , VirulenciaRESUMEN
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.
Asunto(s)
Antibacterianos/farmacología , Caries Dental/microbiología , Bacterias Grampositivas/efectos de los fármacos , Fitoterapia/métodos , Extractos Vegetales/farmacología , Actinomyces/efectos de los fármacos , Annonaceae/química , Arecaceae/química , Brasil , Combretaceae/química , Croton/química , Humanos , Jatropha/química , Lactobacillus acidophilus/efectos de los fármacos , Malpighiaceae/química , Melastomataceae/química , Pruebas de Sensibilidad Microbiana , Fenoles/análisis , Extractos Vegetales/química , Extracto de Senna/química , Solventes/química , Streptococcus gordonii/efectos de los fármacos , Streptococcus mitis/efectos de los fármacos , Streptococcus mutans/efectos de los fármacos , Streptococcus sanguis/efectos de los fármacos , Streptococcus sobrinus/efectos de los fármacos , Temperatura , Terpenos/análisisRESUMEN
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.
Asunto(s)
Adhesión Bacteriana/fisiología , Biopelículas/crecimiento & desarrollo , Boca/microbiología , Soportes Ortodóncicos/microbiología , Streptococcus/fisiología , Óxido de Aluminio/química , Técnicas Bacteriológicas , Cerámica/química , Colorantes , Resinas Compuestas/química , Medios de Cultivo , Materiales Dentales/química , Placa Dental/microbiología , Aleaciones de Oro/química , Humanos , Ensayo de Materiales , Cemento de Policarboxilato/química , Saliva/microbiología , Acero Inoxidable/química , Streptococcus/clasificación , Streptococcus gordonii/fisiología , Streptococcus mutans/fisiología , Streptococcus oralis/fisiología , Streptococcus sanguis/fisiología , Streptococcus sobrinus/fisiología , Sales de Tetrazolio , Tiazoles , Titanio/químicaRESUMEN
BACKGROUND AND OBJECTIVE: Antimicrobial agents provide valuable adjunctive therapy for the prevention and the control of oral diseases. Limitations in their prolonged use have stimulated the search for new, naturally occurring agents with more specific activity and fewer adverse effects. Here we sought to determine the antibacterial properties of blackberry extract (BBE) in vitro against oral bacterial commensals and periodontopathogens. MATERIAL AND METHODS: The effects of whole and fractionated BBE on the metabolism of 10 different oral bacteria were evaluated using the colorimetric water-soluble tetrazolium-1 assay. The bactericidal effects of whole BBE against Fusobacterium nucleatum were determined by quantitating the numbers of colony-forming units (CFUs). Cytotoxicity was determined in oral epithelial (OKF6) cells. RESULTS: BBE at 350-1400 µg/mL reduced the metabolic activity of Porphyromonas gingivalis, F. nucleatum and Streptococcus mutans. The reduced metabolic activity observed for F. nucleatum corresponded to a reduction in the numbers of CFUs following exposure to BBE for as little as 1 h, indicative of its bactericidal properties. An anthocyanin-enriched fraction of BBE reduced the metabolic activity of F. nucleatum, but not of P. gingivalis or S. mutans, suggesting the contribution of species-specific agents in the whole BBE. Oral epithelial cell viability was not reduced following exposure to whole BBE (2.24-1400 µg/mL) for ≤ 6 h. CONCLUSION: BBE alters the metabolic activity of oral periodontopathogens while demonstrating a minimal effect on commensals. The specific antibacterial properties of BBE shown in this study, along with its previously demonstrated anti-inflammatory and antiviral properties, make this natural extract a promising target as an adjunct for prevention and/or complementary therapy of periodontal infections.
Asunto(s)
Antibacterianos/farmacología , Bacterias/efectos de los fármacos , Enfermedades Periodontales/microbiología , Extractos Vegetales/farmacología , Rosaceae , Actinomyces/efectos de los fármacos , Aggregatibacter actinomycetemcomitans/efectos de los fármacos , Antocianinas/farmacología , Antiinfecciosos Locales/farmacología , Carga Bacteriana/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Clorhexidina/análogos & derivados , Clorhexidina/farmacología , Colorimetría/métodos , Células Epiteliales/efectos de los fármacos , Fusobacterium nucleatum/efectos de los fármacos , Humanos , Indicadores y Reactivos , Queratinocitos/efectos de los fármacos , Ensayo de Materiales , Mucosa Bucal/citología , Mucosa Bucal/efectos de los fármacos , Porphyromonas gingivalis/efectos de los fármacos , Prevotella intermedia/efectos de los fármacos , Streptococcus gordonii/efectos de los fármacos , Streptococcus mutans/efectos de los fármacos , Streptococcus oralis/efectos de los fármacos , Sales de Tetrazolio , Factores de Tiempo , Veillonella/efectos de los fármacosRESUMEN
INTRODUCTION: Many pathogenic microorganisms were found in an infected root canal. The object of this study was to evaluate the effect of MTAD in combination with nisin on the pathogens associated with root canal infection. METHODS: The survival rates of 9 pathogenic bacteria were determined after 1-, 5-, and 10-minute treatment with MTAD, MTAN (substitution of doxycycline with nisin), and MTADN (nisin in combination with doxycycline). The survival rates of Enterococcus faecalis in the starvation phase and pretreatment alkalization as well as in the normal physiological state under MTAD, MTAN, and MTADN challenge for 1, 5, and 10 minutes were evaluated and compared. Furthermore, scanning electron microscopy was used to observe the morphologic modification of Actinomyces naeslundii, Lactobacillus paracasei, and Porphyromonas gingivalis after MTAD and MTADN treatment. RESULTS: L. fermenti, L. paracasei, A. viscosus, A. naeslundii, Streptococcus gordonii, and Peptostreptococcus were more sensitive to MTADN and MTAN than to MTAD. MTAD, MTAN, and MTADN showed a rapid antibacterial effect on P. gingivalis, Prevotella intermedia, and Fusobacterium nucleatum. Enterococcus faecalis in the stress state was as sensitive to MTAD, MTAN, and MTADN as the control E. faecalis. Furthermore, in the observation of scanning electron microscopy, the membranes in A. naeslundii and L. paracasei presented significant rupture, and P. gingivalis did not exhibit significant damage after MTADN treatment. CONCLUSIONS: MTAD in combination with nisin improved antibacterial efficacy against pathogens, especially for some gram-positive bacteria associated with persistent intracanal infection. Therefore, the combination had the potential to be used as an effective intracanal irrigation.
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Antibacterianos/farmacología , Ácido Cítrico/farmacología , Enfermedades de la Pulpa Dental/microbiología , Doxiciclina/farmacología , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Nisina/farmacología , Polisorbatos/farmacología , Irrigantes del Conducto Radicular/farmacología , Actinomyces/efectos de los fármacos , Actinomyces viscosus/efectos de los fármacos , Fenómenos Fisiológicos Bacterianos/efectos de los fármacos , Membrana Celular/efectos de los fármacos , Evaluación Preclínica de Medicamentos , Enterococcus faecalis/efectos de los fármacos , Fusobacterium nucleatum/efectos de los fármacos , Humanos , Lactobacillus/efectos de los fármacos , Limosilactobacillus fermentum/efectos de los fármacos , Viabilidad Microbiana/efectos de los fármacos , Microscopía Electrónica de Rastreo , Peptostreptococcus/efectos de los fármacos , Porphyromonas gingivalis/efectos de los fármacos , Prevotella intermedia/efectos de los fármacos , Streptococcus gordonii/efectos de los fármacos , Factores de TiempoRESUMEN
In the oral cavity, omnipresent salivary protein films (pellicle) mediate bacterial adhesion and biofilm formation on natural tissues as well as on artificial implant surfaces, which may cause serious infectious diseases like periimplantitis. The purpose of this in vitro study was to investigate the adsorption/desorption behaviour of human saliva on model surfaces grafted with polyamidoamine (PAMAM) dendrimer molecules compared to self-assembled monolayers (SAMs) exhibiting the same terminal functions (-NH(2), -COOH) by two complementary analytical methods. Furthermore, the role of saliva conditioning of PAMAM and analogous SAM modifications on the adhesion of Streptococcus gordonii DL1, an early oral colonizer, was investigated. In contrast to SAMs, PAMAM-grafted surfaces showed reduced streptococcal adherence in the absence of pre-adsorbed saliva similar to the level obtained for poly(ethylene glycol) (PEG) coatings. Moreover, coatings of PAMAM-NH(2) maintained their bacteria-repellent behaviour even after saliva-conditioning. As a general outcome, it was found that lower amounts of protein adsorbed on PAMAM coatings than on analogous SAMs. Since this study demonstrates that covalently bound PAMAM dendrimers can modulate the oral bacterial response, this approach has significant potential for the development of anti-adhesive biomaterial surfaces that are conditioned with proteinaceous films.
Asunto(s)
Adhesión Bacteriana/efectos de los fármacos , Dendrímeros/química , Dendrímeros/farmacología , Modelos Químicos , Proteínas y Péptidos Salivales/metabolismo , Silicio/farmacología , Streptococcus gordonii/fisiología , Adsorción/efectos de los fármacos , Humanos , Concentración de Iones de Hidrógeno/efectos de los fármacos , Punto Isoeléctrico , Microscopía de Fuerza Atómica , Tecnicas de Microbalanza del Cristal de Cuarzo , Electricidad Estática , Streptococcus gordonii/citología , Streptococcus gordonii/efectos de los fármacos , Propiedades de Superficie/efectos de los fármacos , Agua/químicaRESUMEN
OBJECTIVE: To examine the effect of garlic extract on the biofilm formation by Streptococcus mutans on orthodontic wire and on glucosyltransferase gene expression. MATERIALS AND METHODS: Growth inhibition of oral bacteria was tested after 50 µL of garlic extract was placed on an agar plate. The minimum inhibitory concentration (MIC) of garlic extract on S mutans growth was first determined. After cultivating streptococci in biofilm medium (BM)-sucrose with garlic extract and orthodontic wire, adenosine triphosphate (ATP) measurement and viable cell counting was performed from the bacteria attached on the wire. Scanning electron microscopy (SEM) analysis of morphology was observed on bacterial cells attached to orthodontic wire. The effect of garlic extract on gene expression was evaluated using quantitative real-time polymerase chain reaction (PCR) of glucosyltransferase. RESULTS: Though garlic extract had a clear antibacterial effect on all microorganisms, it also enhanced S mutans attachment on orthodontic wire. Low concentration of garlic extract also increased glucosyltransferase gene expression of S mutans. CONCLUSIONS: Despite its antibacterial function, garlic extract increases biofilm formation by S mutans to orthodontic wire, likely through upregulation of glucosyltransferase expression. Garlic extract may thus play an important role in increased bacterial attachment to orthodontic wires.