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1.
Planta Med ; 85(16): 1242-1252, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31597167

RESUMO

Numerous chemically synthesized compounds are widely used in oral hygiene products. However, due to their potential risk, there is a need to improve the safety and quality of dental care by seeking alternative control agents such as those naturally found in plant materials. Here we assessed antibacterial potentials of extracts from 100 species of Korean native plants against Streptococcus mutans on cariogenesis. Among those, extracts from five plants (Arctii Fructus, Caryopteris incana, Aralia continentalis, Symplocarpus renifolius, and Lamium amplexicaule) showed a growth inhibition of S. mutans. The five extracts were further individually evaluated for their minimal inhibitory concentration and minimal bactericidal concentration. Interestingly, a synergistic antibacterial activity was observed with the combination of sodium fluoride and the plant extracts. To determine the anti-biofilm activity of plant extracts, S. mutans was treated with increasing concentrations of the extracts in the range from 1250 to 3750 µg/mL. When S. mutans was grown in the defined biofilm medium containing the individual extracts of 47 species, the biofilm amount markedly decreased compared to that of a negative control. Notably, the extract of S. renifolius significantly downregulated the gtf and spaP genes for synthesis of glucan and adhesive proteins in S. mutans, and L. amplexicaule decreased the expression of gtfD gene. Therefore, these results demonstrate that the five plant extracts modulate survival and pathogenesis of S. mutans by growth inhibition and downregulation of the gene(s) implicated in biofilm formation.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Produtos Biológicos/farmacologia , Streptococcus mutans/efeitos dos fármacos , Antibacterianos/química , Produtos Biológicos/química , República da Coreia , Streptococcus mutans/crescimento & desenvolvimento
2.
Int J Mol Sci ; 20(19)2019 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-31547104

RESUMO

Antibacterial resistance (ABR) poses an enormous threat to human health. ABR mainly develops due to bacteria being constantly exposed to diluted levels of disinfectants. Here, we propose a method for suppressing ABR through the chemical binding of disinfectants to polymethyl methacrylate (PMMA) device surfaces in solutions of 5%, 10%, and 20% disinfectant concentrations. PMMA discs were fabricated from a commercial orthodontic acrylic resin system (Ortho-Jet) and quaternary ammonium salts (QAS), 3-(trimethoxysilyl)-propyldimethyloctadecyl ammonium chloride (42% in methanol), were used as the disinfectant. The PMMA surfaces were activated in 3 M sulfuric acid at 80 °C for 5 h for the esterification of hydrolyzed QAS to PMMA. Fourier transform infrared difference spectra confirmed that the carboxy-terminated PMMA was chemically bound to the QAS. In vitro cell viability tests using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays revealed that 5%QAS-c-PMMA was more biocompatible than 10%QAS-c-PMMA and 20%QAS-c-PMMA. The results of antibacterial tests and clinical trials demonstrated the excellent antibacterial power of 5%QAS-c-PMMA. This method is the first solution-based approach to successfully avoid disinfectant leakage and subsequent ABR, as revealed by mass spectrometry studies of the solution obtained by agitating the disinfectant-bound PMMA for 28 days.


Assuntos
Desinfetantes , Farmacorresistência Bacteriana/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Fibroblastos/metabolismo , Polimetil Metacrilato , Compostos de Amônio Quaternário , Streptococcus mutans/crescimento & desenvolvimento , Sobrevivência Celular/efeitos dos fármacos , Desinfetantes/química , Desinfetantes/farmacologia , Humanos , Polimetil Metacrilato/química , Polimetil Metacrilato/farmacologia , Compostos de Amônio Quaternário/química , Compostos de Amônio Quaternário/farmacologia
3.
J Appl Biomater Funct Mater ; 17(3): 2280800019847067, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31530071

RESUMO

BACKGROUND: Biofilm formation and microbial colonization on the surface of implant devices may cause dental caries and peri-implantitis. Therefore, various surface treatments have been developed to improve the antibacterial activity of titanium implant. METHODS: Silver-loaded polydopamine coating was formed by immersing pure titanium in dopamine hydrochloride/HCl buffer solution for 24 h in 50 mL silver nitrate solutions with different concentrations for 30 min. Microbial growth inhibition and microbial growth curve analyses for bacterial solutions of Streptococcus mutans and Porphyromonas gingivalis incubated with the specimens were respectively conducted by counting the numbers of colonies on agar solid medium and by measuring absorbance using enzyme-linked immunosorbent assay reader. RESULTS: Silver nanoparticles were uniformly distributed over the whole surface of the polydopamine and silver-coated titanium specimens. The numbers of microbial colonies for both bacteria cultured with surface-modified titanium were significantly lower than those cultured with uncoated titanium. When Streptococcus mutans and Porphyromonas gingivalis were cultured with surface-modified titanium, the lag phase of the growth curves for both bacteria was continually maintained, whereas the lag phase for Streptococcus mutans and Porphyromonas gingivalis changed to exponential phase after 9 and 15 h, respectively, when both bacteria were cultured with uncoated titanium. CONCLUSION: It was confirmed that the coating of polydopamine and silver on the surface of titanium effectively retards the microbial growth, which can cause the formation of biofilm and pathogenesis of gum disease in the mouth.


Assuntos
Antibacterianos , Materiais Revestidos Biocompatíveis , Implantes Dentários/microbiologia , Indóis , Polímeros , Porphyromonas gingivalis/crescimento & desenvolvimento , Streptococcus mutans/crescimento & desenvolvimento , Titânio , Antibacterianos/química , Antibacterianos/farmacologia , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia , Humanos , Indóis/química , Indóis/farmacologia , Nanopartículas Metálicas/química , Polímeros/química , Polímeros/farmacologia , Prata/química , Prata/farmacologia , Titânio/química , Titânio/farmacologia
4.
ACS Appl Mater Interfaces ; 11(41): 37381-37396, 2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31517483

RESUMO

Biodegradable synthetic scaffolds hold great promise for oral and craniofacial guided tissue regeneration and bone regeneration. To overcome the limitations of current scaffold materials in terms of osteogenic and antimicrobial properties, we have developed a novel silver-modified/collagen-coated electrospun poly-lactic-co-glycolic acid/polycaprolactone (PLGA/PCL) scaffold (PP-pDA-Ag-COL) with improved antimicrobial and osteogenic properties. Our novel scaffold was generated by electrospinning a basic PLGA/PCL matrix, followed by silver nanoparticles (AgNPs) impregnation via in situ reduction, polydopamine coating, and then coating by collagen I. The three intermediate materials involved in the fabrication of our scaffolds, namely, PLGA/PCL (PP), PLGA/PCL-polydopamine (PP-pDA), and PLGA/PCL-polydopamine-Ag (PP-pDA-Ag), were used as control scaffolds. Scanning electron micrographs and mechanical testing indicated that the unique three-dimensional structures with randomly oriented nanofibrous electrospun scaffold architectures, the elasticity modulus, and the tensile strength were maintained after modifications. CCK-8 cell proliferation analysis demonstrated that the PP-pDA-Ag-COL scaffold was associated with higher MC3T3 proliferation rates than the three control scaffolds employed. Scanning electron and fluorescence light microscopy illustrated that PP-pDA-Ag-COL scaffolds significantly enhanced MC3T3 cell adhesion compared to the control scaffolds after 12 and 24 h culture, in tandem with the highest ß1 integrin expression levels, both at the mRNA level and the protein level. Alkaline phosphatase activity, BMP2, and RUNX2 expression levels of MC3T3 cells cultured on PP-pDA-Ag-COL scaffolds for 7 and 14 days were also significantly higher when compared to controls (P < 0.001). There was a wider antibacterial zone associated in PP-pDA-Ag-COL and PP-pDA-Ag scaffolds versus control scaffolds (P < 0.05), and bacterial fluorescence was reduced on the Ag-modified scaffolds after 24 h inoculation against Staphylococcus aureus and Streptococcus mutans. In a mouse periodontal disease model, the PP-pDA-Ag-COL scaffold enhanced alveolar bone regeneration (31.8%) and was effective for periodontitis treatment. These results demonstrate that our novel PP-pDA-Ag-COL scaffold enhanced biocompatibility and osteogenic and antibacterial properties and has therapeutic potential for alveolar/craniofacial bone regeneration.


Assuntos
Antibacterianos , Regeneração Óssea/efeitos dos fármacos , Materiais Revestidos Biocompatíveis , Teste de Materiais , Osteogênese/efeitos dos fármacos , Periodontite/terapia , Staphylococcus aureus/crescimento & desenvolvimento , Streptococcus mutans/crescimento & desenvolvimento , Tecidos Suporte/química , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Linhagem Celular , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia , Colágeno/química , Colágeno/farmacologia , Modelos Animais de Doenças , Regeneração Tecidual Guiada , Camundongos , Periodontite/patologia , Poliésteres/química , Poliésteres/farmacologia , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/farmacologia , Prata/metabolismo , Prata/farmacologia
5.
Arch Oral Biol ; 107: 104484, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31382161

RESUMO

OBJECTIVES: The aim of this study was to examine the effect of nano-hydroxyapatite (nHA) on biofilm formation by Streptococcus mutans, which is actively involved in the initiation of dental caries. DESIGN: The effects of nHA on growth and biofilm formation by S. mutans were investigated in two media: a saliva analog medium, basal medium mucin (BMM); and a nutrient-rich medium, brain heart infusion (BHI); in the presence and absence of sucrose. RESULTS: Sucrose enhanced the growth of S. mutans in both media. In the presence of sucrose, nHA enhanced bacterial growth and biofilm formation more in BMM medium than in BHI. nHA also affected the transcription of glucosyltransferase (gtf) genes and production of polysaccharide differently in the two media. In BHI medium, the transcription of all three gtf genes, coding for enzymes that synthesize soluble and insoluble glucans from sucrose, was increased more than 3-fold by nHA. However, in BMM medium, only the transcription of gtfB and gtfC, coding for insoluble glucans, was substantially enhanced by nHA. CONCLUSIONS: nHA appeared to enhance biofilm formation by increasing glucosyltransferase transcription, which resulted in an increase in production of insoluble glucans. This effect was influenced by the growth conditions.


Assuntos
Biofilmes/efeitos dos fármacos , Durapatita/farmacologia , Streptococcus mutans/crescimento & desenvolvimento , Meios de Cultura/química , Cárie Dentária , Glucosiltransferases , Nanopartículas , Streptococcus mutans/efeitos dos fármacos
6.
Microsc Res Tech ; 82(11): 1869-1877, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31373738

RESUMO

Laser irradiation has been proposed as a preventive method against dental caries since it is capable to inhibit enamel demineralization by reducing carbonate and modifying organic matter, yet it can produce significant morphological changes. The purpose of this study was to evaluate the influence of Er:YAG laser irradiation on superficial roughness of deciduous dental enamel and bacterial adhesion. Fifty-four samples of deciduous enamel were divided into three groups (n = 18 each). G1_control (nonirradiated); G2_100 (7.5 J/cm2 ) and G3_100 (12.7 J/cm2 ) were irradiated with Er:YAG laser at 7.5 and 12.7 J/cm2 , respectively, under water irrigation. Surface roughness was measured before and after irradiation using a profilometer. Afterwards, six samples per group were used to measure bacterial growth by XTT cell viability assay. Adhered bacteria were observed using confocal laser scanning microscopy (CLSM) and a scanning electron microscopy (SEM). Paired t-, one-way analysis of variance (ANOVA), Kruskal-Wallis and pairwise Mann-Whitney U tests were performed to analyze statistical differences (p < .05). Before treatment, samples showed homogenous surface roughness, and after Er:YAG laser irradiation, the surfaces showed a significant increase in roughness values (p < .05). G3_100 (12.7 J/cm2 ) showed the highest amount of Streptococcus mutans adhered (p < .05). The increase in the roughness of the tooth enamel surfaces was proportional to the energy density used; the increase in surface roughness caused by laser irradiation did not augment the adhesion of Streptococcus sanguinis; only the use of the energy density of 12.7 J/cm2 favored significantly the adhesion of S. mutans.


Assuntos
Aderência Bacteriana/efeitos da radiação , Cárie Dentária/prevenção & controle , Esmalte Dentário/efeitos da radiação , Lasers de Estado Sólido/uso terapêutico , Streptococcus mutans/fisiologia , Streptococcus/fisiologia , Aderência Bacteriana/fisiologia , Humanos , Microscopia Eletrônica de Varredura , Streptococcus/crescimento & desenvolvimento , Streptococcus/efeitos da radiação , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus mutans/efeitos da radiação , Propriedades de Superfície/efeitos da radiação
7.
Curr Microbiol ; 76(11): 1238-1246, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31377819

RESUMO

Streptococcus mutans (S. mutans) adheres to the tooth surface, metabolizes carbohydrates, and produces acid products, leading to enamel demineralization-the onset of dental caries. Rapid acidification by S. mutans has been observed in the presence of glucose. However, little is known about the role of small RNAs (sRNAs) in S. mutans in the presence of glucose and their relationship to tooth adherence. The objective of this study was to evaluate the role of sRNAs in S. mutans (18-50 nucleotides) regarding adherence capacity under 1% and 5% glucose concentrations. The pH drop and adherence capacity in the 1% glucose condition were similar to these parameters under conditions of 5% sucrose that were published in our previous study. A total of 2149 candidate sRNA with at least 100 average reads in the 5% and 1% glucose libraries were obtained. Between the two libraries, 581 sRNAs were differentially expressed and 43 sRNAs were verified. However, the expression levels of the predicted target genes gtfC and spaP were similar between the 1% and 5% glucose conditions. The bioinformatic analysis suggested that differentially expressed sRNAs may be involved in several pathways. These findings indicate that sRNAs were induced under these glucose concentrations and a series of sRNAs were specifically induced, respectively. sRNAs that are induced under glucose stress may be involved in regulating adherence of S. mutans.


Assuntos
Aderência Bacteriana , Glucose/análise , RNA Bacteriano/metabolismo , Streptococcus mutans/fisiologia , Cárie Dentária/microbiologia , Regulação Bacteriana da Expressão Gênica , Glucose/metabolismo , Humanos , RNA Bacteriano/genética , Streptococcus mutans/genética , Streptococcus mutans/crescimento & desenvolvimento , Sacarose/metabolismo
8.
Colloids Surf B Biointerfaces ; 182: 110359, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31352253

RESUMO

Multiple ions codoping may effectively modulate physicochemical and biological properties of hydroxyapatite (HA) for diverse biomedical applications. This study synthesized strontium (Sr)-, fluorine (F)- doped, and Sr/F-codoped HA nanoparticles by a hydrothermal method, and investigated the effect of ion doping on characteristics of HA, including crystallinity, crystal size, lattice parameters, and substitution sites by experiments and simulation with density functional theory (DFT) methods. It was found that, Sr doping increased the lattice parameters of HA whereas F doping decreased these parameters. Additionally, F doping enhanced the structural stability of the Sr-doped HA. F doping created excellent antibacterial properties to effectively inhibit growth of Streptococcus mutans. An appropriate Sr doping level endowed HA with optimum osteogenic ability to promote osteoblastic differentiation of bone marrow stem cells. These suggest that, Sr/F codoping is an effective approach to synthesizing HA-based materials with both antibacterial and osteogenic properties. More broadly, HA nanomaterials with specific characteristics may be designed for meeting diverse requirements from biomedical applications.


Assuntos
Antibacterianos/síntese química , Durapatita/química , Fluoretos/química , Células-Tronco Mesenquimais/efeitos dos fármacos , Nanopartículas/química , Osteoblastos/efeitos dos fármacos , Estrôncio/química , Antibacterianos/farmacologia , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Cristalização , Durapatita/farmacologia , Fluoretos/farmacologia , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/fisiologia , Testes de Sensibilidade Microbiana , Estrutura Molecular , Osteoblastos/citologia , Osteoblastos/fisiologia , Osteogênese/efeitos dos fármacos , Teoria Quântica , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/crescimento & desenvolvimento , Estrôncio/farmacologia , Engenharia Tecidual
9.
Microbiol Immunol ; 63(7): 251-260, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31166029

RESUMO

Streptococcus mutans is a cariogenic bacterium that localizes in the oral cavity. Glycyrrhetinic acid (GRA) is a major component of licorice extract. GRA and several derivatives, including disodium succinoyl glycyrrhetinate (GR-SU), are known to have anti-inflammatory effects in humans. In this study, the antimicrobial effect of GRA and its derivatives against the S. mutans UA159 strain were investigated. Minimum inhibitory concentrations (MICs) of GRA and GR-SU showed antibacterial activity against the S. mutans strain, whereas other tested derivatives did not. Because GR-SU is more soluble than GRA, GR-SU was used for further experiments. The antibacterial activity of GR-SU against 100 S. mutans strains was evaluated and it was found that all strains are susceptible to GR-SU, with MIC values below 256 µg/mL. A cell viability assay showed that GR-SU has a bacteriostatic effect on S. mutans cells. As to growth kinetics, sub-MICs of GR-SU inhibited growth. The effect of GR-SU on S. mutans virulence was then investigated. GR-SU at sub-MICs suppresses biofilm formation. Additionally, GR-SU greatly suppresses the pH drop caused by the addition of glucose and glucose-induced expression of the genes responsible for acid production (ldh and pykF) and tolerance (aguD and atpD). Additionally, expression of enolase, which is responsible for the carbohydrate phosphotransferase system, was not increased in the presence of GR-SU, indicating that GR-SU suppresses incorporation of sugars into S. mutans. In conclusion, GR-SU has antibacterial activity against S. mutans and also decreases S. mutans virulence.


Assuntos
Antibacterianos/farmacologia , Ácido Glicirretínico/farmacologia , Glycyrrhiza/química , Extratos Vegetais/farmacologia , Streptococcus mutans/efeitos dos fármacos , Antibacterianos/química , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Genes Bacterianos/genética , Glucose/metabolismo , Ácido Glicirretínico/química , Concentração de Íons de Hidrogênio , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Extratos Vegetais/química , Streptococcus mutans/genética , Streptococcus mutans/crescimento & desenvolvimento , Virulência , Fatores de Virulência/genética , Fatores de Virulência/metabolismo
10.
J Agric Food Chem ; 67(26): 7297-7303, 2019 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-31244198

RESUMO

3,6-Anhydro-l-galactose (AHG) produced from agarose in red macroalgae was recently suggested as an anticariogenic sugar to replace widely used xylitol. However, the multi-step process for obtaining monomeric sugar AHG from agarose may be expensive. Generally, it is easier to obtain oligosaccharides than monosaccharides from polysaccharides. Therefore, a one-step process to obtain agarobiose (AB) from agarose was recently developed, and here, we suggest AB as a new anticariogenic agent, owing to its anticariogenic activity against Streptococcus mutans. Among AHG-containing oligosaccharides, AB, neoagarobiose (NAB), agarooligosaccharides (AOSs), and neoagarooligosaccharides (NAOSs), AB showed higher inhibitory activity than AOSs against the growth and lactic acid production of S. mutans; no such inhibitory activity was observed for NAB and NAOSs. This inhibitory effect of AB was comparable to the previously reported inhibitory activity of AHG against S. mutans. These results suggest that AB, which can be more economically and simply produced than AHG, may serve as an anticariogenic sugar.


Assuntos
Antibacterianos/farmacologia , Dissacarídeos/farmacologia , Oligossacarídeos/farmacologia , Extratos Vegetais/farmacologia , Rodófitas/química , Alga Marinha/química , Antibacterianos/química , Antibacterianos/isolamento & purificação , Dissacarídeos/química , Dissacarídeos/isolamento & purificação , Oligossacarídeos/química , Oligossacarídeos/isolamento & purificação , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/crescimento & desenvolvimento
11.
Artigo em Inglês | MEDLINE | ID: mdl-31041198

RESUMO

Streptococcus oralis subspecies dentisani is explored as an anti-cariogenic probiotic. Here, subjecting freshly stimulated saliva samples of 35 healthy volunteers, six epidemiologically unrelated and two related strains were isolated (prevalence around 20%) applying a newly developed three-step procedure. Furthermore, the probiotic strain S. dentisani 7746 (AB-Dentisanium®) was tested under a variety of environmental conditions for its inhibitory effect on six S. mutans, two S. sobrinus, 15 other oral or intestinal streptococci, 15 S. dentisani strains, and six representatives of other species including periodontopathogens. All except one of the S. mutans strains were inhibited by 7746 colonies or culture supernatant concentrate but only if either the test cell number was low or the producer or its bacteriocin concentration, respectively, was high. S. sanguinis OMI 332, S. salivarius OMI 315, S. parasanguinis OMI 335, S. vestibularis OMI 238, and the intestinal S. dysgalactiae OMI 339 were not inhibited, while the other 10 streptococcal strains (especially S. oralis OMI 334 and intestinal S. gallolyticus OMI 326) showed a certain degree of inhibition. From the panel of other bacterial species only Aggregatibacter actinomycetemcomitans was slightly inhibited. With the exception of OMI 285 and OMI 291 that possessed a 7746 bacteriocin-like gene cluster, all S. dentisani strains and especially type strain 7747T were strongly inhibited by 7746. In conclusion, probiotic strain 7746 might antagonize the initiation and progression of dental caries by reducing S. mutans if not too abundant. S. dentisani strains inhibit each other, but strains with similar bacteriocin-related gene clusters, including immunity genes, are able to co-exist due to cross-resistance. In addition, development of resistance and adaptation to 7746-bacteriocins was observed during our study and needs attention. Hence, mechanisms underlying such processes need to be further investigated using omics-approaches. On the manufacturing level, probiotic strains should be continuously tested for function. Further clinical studies investigating inhibition of S. mutans by AB-Dentisanium® are required that should also monitor the impact on the oral microbiome composition including resident S. dentisani strains.


Assuntos
Antibacterianos/metabolismo , Técnicas de Tipagem Bacteriana/métodos , Bacteriocinas/metabolismo , Probióticos/isolamento & purificação , Infecções Estreptocócicas/microbiologia , Streptococcus oralis/classificação , Streptococcus oralis/isolamento & purificação , Aggregatibacter actinomycetemcomitans/efeitos dos fármacos , Antibiose , Portador Sadio/epidemiologia , Portador Sadio/microbiologia , Voluntários Saudáveis , Prevalência , Infecções Estreptocócicas/epidemiologia , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus oralis/efeitos dos fármacos , Streptococcus sobrinus/crescimento & desenvolvimento
12.
Microb Pathog ; 133: 103546, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31112769

RESUMO

With this study, we investigated the effect of synthetic antimicrobial peptides Pep19-2.5 and Pep194LF alone or in combination with antibiotics on S. mutans growth and biofilm formation/disruption. We also examined the cytotoxic effect of each peptide on monocytes. S. mutans was cultured in the presence of different concentrations of each peptide. We showed that Pep19-2.5 and Pep19-4LF were able to significantly (p ≤ 0.01) inhibit the growth of S. mutans. The synthetic peptides also decreased biofilm formation by S. mutans. Furthermore, both peptides reduced the viability of S. mutans in already formed biofilms. The combination of each peptide with antibiotics (penicillin/streptomycin, P/S) produced additive interactions which inhibited S. mutans growth and biofilm formation. Pep19-2.5 and Pep19-4LF were nontoxic, as they did not decrease monocyte viability and did not increase the lactate dehydrogenase activity of the exposed cells. In conclusion, synthetic peptides Pep19-2.5 and Pep19-4LF did inhibit S. mutans growth and its capacity to form biofilm. Both peptides were found to be nontoxic to monocytes. These data provide new insight into the efficacy of synthetic peptides Pep19-2.5 and Pep19-4LF against S. mutans. These peptides may thus be useful in controlling the adverse effects of this cariogenic bacterium in human.


Assuntos
Anti-Infecciosos/farmacologia , Biofilmes/efeitos dos fármacos , Proteínas do Tecido Nervoso/farmacologia , Peptídeos/farmacologia , Streptococcus mutans/efeitos dos fármacos , Peptídeos Catiônicos Antimicrobianos , Biofilmes/crescimento & desenvolvimento , Sobrevivência Celular/efeitos dos fármacos , Combinação de Medicamentos , Sinergismo Farmacológico , Humanos , L-Lactato Desidrogenase/metabolismo , Testes de Sensibilidade Microbiana , Monócitos/efeitos dos fármacos , Proteínas do Tecido Nervoso/síntese química , Penicilinas/farmacologia , Peptídeos/síntese química , Streptococcus mutans/crescimento & desenvolvimento
13.
J Ethnopharmacol ; 239: 111872, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-30991136

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Chronic periodontitis is an inflammatory process of the oral cavity that is caused by different reasons. Streptococcus mutans bacteria is the main cause of the disease. Scrophularia striata in Iranian traditional medicine is used to heal inflammation of gum and mouth. Also, In vitro anti-bacterial effect of S. striata in S. mutans have been demonstrated. AIM OF THE STUDY: Clinical, in vitro and chemical constituent studies of S. striata on chronic periodontitis disease. MATERIALS AND METHODS: This study is a randomized clinical trial that was performed in Dental College, Hamadan University of Medical Sciences. Fifty patients with chronic periodontitis (20-50 years old) were selected and the study was approved and registered in Iranian Registry of Clinical Trials as number IRCT2015081323468N1 prior to the experiment. The 50 patients were divided into two groups. One group used herbal mouthwash that was made from hydro alcoholic extract of S. striata and another group used Irsha mouthwash (Iranian form of Listerine mouthwash). Patients were followed up after two weeks and four weeks of using mouthwash for assessment of plaque index (PI), pocket depth (PD), bleeding on probing (BOP) and counting Streptococcus mutans. The mouthwash was standardized according to gallic acid, quercetin and apigenin reference using the RP-HPLC method. The total phenolic and flavonoids contents were also determined spectrophotometerically. RESULTS: S. striata in the short term, improve plaque index, pocket depth and bleeding on probing but the number of Streptococcus mutans changed significantly in the long term and is more potent in comparison to Irsha mouthwash. The mouthwash was standardized according to gallic acid, quercetin and apigenin standard (3, 24, 11 µg, respectively). The total phenolic and flavonoid content of mouthwash were 120.7 µg GAE/100 cc and 78.6 µg QE/100 cc respectively. CONCLUSIONS: S. striata mouthwash is effective in the treatment of chronic periodontitis disease and is more potent comparing the Iranian Irsha mouthwash.


Assuntos
Periodontite Crônica/tratamento farmacológico , Antissépticos Bucais/uso terapêutico , Extratos Vegetais/uso terapêutico , Scrophularia , Adulto , Índice de Placa Dentária , Feminino , Hemorragia/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-Idade , Antissépticos Bucais/química , Compostos Fitoquímicos/análise , Compostos Fitoquímicos/uso terapêutico , Extratos Vegetais/química , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/crescimento & desenvolvimento , Resultado do Tratamento , Adulto Jovem
14.
J Clin Pediatr Dent ; 43(3): 181-184, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30964724

RESUMO

Objectives: To compare the in vitro cariogenic potential of two infant formulas (IF, for ages 6-12 months) processed with sterile distilled water with or without 1 ppm fluoride. Study design: Nine specimens in each water type were inoculated with Streptococcus mutans (SM) suspension. The specimens were then divided into one group of samples cultured immediately on Mitis salivarius (MS) agar plates (T0) and another group of specimens cultured on MS agar plates after incubation at 37°C for 4 hours in anaerobic conditions (T4). Six-fold dilutions of each sample were incubated for 48 hours, and colony-forming units (CFUs) of SM were numerated. The pH changes associated with bacterial fermentation of each of the suspensions were measured at T0 and at T4 following incubation. Results: The pH was lower at T4 than at T0 in both IF. The SM colonies increased significantly at T4 compared to T0 in both IF (P < 0.001). There was no significant difference between the CFUs of the mediums at T0 and T4 or relation between SM colonies of IF based on fluoridated and non-fluoridated distilled water. Conclusion: The cariogenic potential was not significantly different between two IF prepared with fluoridated or non-fluoridated distilled water.


Assuntos
Cariogênicos , Fluoretos , Fórmulas Infantis , Streptococcus mutans , Fluoretos/administração & dosagem , Fluoretos/uso terapêutico , Humanos , Lactente , Fórmulas Infantis/efeitos adversos , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/crescimento & desenvolvimento
15.
Arch Biochem Biophys ; 666: 46-51, 2019 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-30930283

RESUMO

Streptococcus mutans, a bacterium mainly inhabiting the tooth surface, is a major pathogen of dental caries. The bacterium metabolizes sugars to produce acids, resulting in an acidic microenvironment in the dental plaque. Hence, S. mutans should possess a mechanism for surviving under acidic conditions. In the current study, we report the effects of inhibitors of Escherichia coli proton-pumping F-type ATPase (F-ATPase) on the activity of S. mutans enzyme, and the growth and survival of S. mutans under acidic conditions. Piceatannol, curcumin, and demethoxycurcumin strongly reduced the ATPase activity of S. mutans F-ATPase. Interestingly, these compounds inhibited the growth of S. mutans at pH 5.3 but not at pH 7.3. They also significantly reduced the colony-forming ability of S. mutans after incubation at pH 4.3, while showing essentially no effect at pH 7.3. These observations indicate that S. mutans is highly sensitive to F-ATPase inhibitors under acidic conditions and that F-ATPase plays an important role in acid tolerance of this bacterium.


Assuntos
Adenosina Trifosfatases/metabolismo , Concentração de Íons de Hidrogênio , Bombas de Próton/metabolismo , Streptococcus mutans/enzimologia , Streptococcus mutans/crescimento & desenvolvimento
16.
J Med Food ; 22(6): 623-630, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31021282

RESUMO

This study was designed to determine whether the ethanol extract of Artemisia princeps could inhibit the cariogenic activity of Streptococcus mutans. The increase in acid production and biofilm formation by S. mutans were evaluated. The expression levels of virulence factor genes were determined by performing the real-time polymerase chain reaction (PCR). The bactericidal effect was tested by confocal laser scanning microscopy. The A. princeps extract was observed to inhibit the growth of S. mutans at concentrations >0.05 mg/mL (P < .05). After using the safranin staining method, we found that the A. princeps extract had an inhibitory effect against biofilm formation at a concentration of >0.05 mg/mL. These experimental results were similar to that observed with the scanning electron microscopy. The results of the confocal microscopy revealed that the A. princeps extract at high concentrations of 0.4-3.2 mg/mL showed a bactericidal effect in a concentration-dependent manner. According to the results of the real-time PCR analysis, it was observed that the A. princeps extract inhibited the expression of virulence factor genes. These results suggest that A. princeps may inhibit the cariogenic activity of S. mutans, and may be useful as an anticariogenic agent.


Assuntos
Antibacterianos/farmacologia , Artemisia/química , Proteínas de Bactérias/genética , Biofilmes/efeitos dos fármacos , Extratos Vegetais/farmacologia , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/genética , Fatores de Virulência/genética , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus mutans/fisiologia , Fatores de Virulência/metabolismo
17.
Carbohydr Polym ; 212: 142-149, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30832841

RESUMO

Mentha piperita essential oils (MPEO) were loaded into chitosan nanogel to use as antibiofilm agent against Streptococcus mutans and to protect its dental plaque. Chitosan nanoparticles (CsNPs) were prepared by sol-gel method using linking bridge of tripolyphosphate (TPP). Physiological properties of MPEO-CNs were assessed by FTIR, SEM/EDX, DLS and zeta potential. Release kinetics, MIC and MBC were determined for MPEO-CNs. Expression of biofilm-associated genes including 8 genes: grfB, C and D, brpA, spaP, gbpB, relA and vicR was investigated at the presence of sub-MIC of MPEO-CNs. Most abundant bioactive compounds of MPEO were l-menthol (45.05%) and l-menthal (17.53%). SEM/EDX exhibited successful entrapment of MPEO into CsNPs followed by the changes in abundance of elemental peaks. A signal at 1737 cm-1 on chitosan spectrum was attributed to the carboxylic (CO) groups overlapped by MPEO incorporation. A new signal at 2361 cm-1 was assigned to electrostatic interactions of amine groups in chitosan with phosphoric units of TPP within the MPEO-chitosan. MPEO incorporation into porous nanogel decreased monodispersity of the nanoparticles and then raises z-average. Maximum release of MPEO was about 50% during 360 h in a hydroalcoholic solvent at ambient temperature. The adherence of bacterial cells showed high sensitivity to the nanoformulation of MPEO compared with unloaded chitosan-nanogel. Antibiofilm inhibition of S. mutans occurred in 50 and 400 µg/mL for MPEO-CNs and unloaded-nanogel, respectively. Among biofilm synthesis genes, gtfB, gtfC, gtfD were slightly affected by MPEO-CNs treatment, while gbpB, spaP, brpA, relA, and vicR genes underwent significant down-regulation in the presence of both unloaded-nanogel and MPEO-loaded-nanogel. This study demonstrated that the MPEO-CNs promised an efficient nanoformulation with the greatest inhibitory action against some glycosyltransferase genes (gtfB, C and D) as important enzymes involved in extracellular polymers. Finally, the results concluded that MPEO-CNs have a potential use as antibiofilm agent in toothpaste or mouth washing formulations.


Assuntos
Biofilmes/efeitos dos fármacos , Quitosana/administração & dosagem , Óleos Vegetais/administração & dosagem , Polietilenoglicóis/administração & dosagem , Polietilenoimina/administração & dosagem , Streptococcus mutans/efeitos dos fármacos , Dente/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Quitosana/metabolismo , Cárie Dentária/tratamento farmacológico , Cárie Dentária/microbiologia , Placa Dentária/tratamento farmacológico , Placa Dentária/microbiologia , Humanos , Mentha piperita , Óleos Vegetais/isolamento & purificação , Óleos Vegetais/metabolismo , Polietilenoglicóis/metabolismo , Polietilenoimina/metabolismo , Streptococcus mutans/crescimento & desenvolvimento , Dente/microbiologia
18.
Molecules ; 24(3)2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30717122

RESUMO

This study aimed to test the effectiveness of ethyl gallate (EG) against S. mutans biofilm formation on solid surfaces (polystyrene, glass) and acidogenicity, and to examine the effect on expression of related genes. The biofilm that is formed by S. mutans bacteria was evaluated using colorimetric assay and optical profilometry, while the pH of the biofilm growth medium was measured with microelectrode. The expression of genes encoding glucan binding protein B (gbpB), glucosyltranferases B, -C, -D (gtfB, -C, -D) and F-ATPase (atpD, atpF) was assessed using a quantitative reverse transcription-polymerase chain reaction (RT-qPCR). It was revealed that all of the EG concentrations significantly suppressed S. mutans biofilm build-up on polystyrene and glass surfaces, and inhibited acidogenicity, in a dose-dependent manner, compared to the activity of untreated bacteria (p < 0.05). The highest concentration of EG (3.53 mM) reduced biofilm formation on polystyrene and glass surfaces by 68% and more than 91%, respectively, and prevented a decrease in pH levels by 95%. The RT-qPCR data demonstrate that the biofilm-producing bacteria treated with EG underwent significant gene expression changes involving the gtfC (a 98.6 increase in fold change), gtfB gene (a 47.5 increase in fold change) and the gbpB gene (a 13.8 increase in fold change). However, for the other genes tested (gtfD, atpD and atpF), the EG treatments did not produce significant expression change compared to the control. EG produced significant gene expression change in three genes-gtfC, gtfB, and gbpB; it has the capacity to inhibit S. mutans biofilm formation on solid surfaces (polystyrene, glass), as well as acidogenicity. Therefore, EG might be used as an antibiofilm and/or anticaries agent for oral formulations in order to reduce the prevalence of dental caries.


Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Biofilmes/efeitos dos fármacos , Ácido Gálico/análogos & derivados , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Streptococcus mutans/efeitos dos fármacos , Aderência Bacteriana/efeitos dos fármacos , Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Meios de Cultura/química , Cárie Dentária/microbiologia , Cárie Dentária/prevenção & controle , Relação Dose-Resposta a Droga , Ácido Gálico/farmacologia , Vidro/química , Glucosiltransferases/genética , Glucosiltransferases/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Lectinas/genética , Lectinas/metabolismo , Testes de Sensibilidade Microbiana , Poliestirenos/química , ATPases Translocadoras de Prótons/genética , ATPases Translocadoras de Prótons/metabolismo , Streptococcus mutans/genética , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus mutans/metabolismo
19.
Molecules ; 24(3)2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30717140

RESUMO

There is no consensus on the antibacterial activity of dentin bonding systems (DBS). Many study models have been used to evaluate the antimicrobial activity of dental materials. In this study, a novel detection method, flow cytometry, was introduced. It allows for evaluation of the antibacterial activity of DBS, based on assessment of the disruption of the bacterial physical membrane induced by DBS. The aim of the study was to evaluate the antibacterial properties of selected dentin bonding systems against Streptococcus mutans. The highest antibacterial activity against S. mutans was observed for Adhese Universal (99.68% dead cells) and was comparable to that of Prime&Bond Universal, OptiBond Universal, or Clearfil Universal Bond Quick (p > 0.05). The lowest activity of all tested systems was displayed by the multi-mode adhesive, Universal Bond (12.68% dead bacteria cells), followed by the self-etch adhesive, OptiBond FL (15.58% dead bacteria cells). The present study showed that in the case of two-component DBS, the primer exhibited higher antimicrobial activity than the adhesive (or bond) itself.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Adesivos Dentinários/farmacologia , Streptococcus mutans/efeitos dos fármacos , Antibacterianos/química , Antibacterianos/classificação , Biofilmes/crescimento & desenvolvimento , Cárie Dentária/microbiologia , Cárie Dentária/prevenção & controle , Adesivos Dentinários/química , Adesivos Dentinários/classificação , Relação Dose-Resposta a Droga , Citometria de Fluxo , Humanos , Teste de Materiais , Testes de Sensibilidade Microbiana , Streptococcus mutans/genética , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus mutans/metabolismo
20.
Mol Oral Microbiol ; 34(2): 51-63, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30659765

RESUMO

The cariogenic pathogen Streptococcus mutans effectively utilizes dietary sucrose for the synthesis of exopolysaccharides (EPS), which act as a scaffold for its biofilm and thus contribute to its cariogenic pathogenicity. Dextranase (Dex), which is a type of glucanase, participates in the degradation of water-soluble glucan (WSG); however, the structural features of the EPS regulated by the dexAgene have received limited attention. Our recent studies reported novel protocols to fractionate and analyzed the structural characteristics of glucans from S mutans biofilms. In this study, we identify the role of the S mutans dexAgene in dextran-dependent aggregation in biofilm formation. Our results show that deletion of dexA (SmudexA) results in increased transcription of EPS synthesis-related genes, including gtfB, gtfD, and ftf. Interestingly, we reveal that inactivating the dexA gene may lead to elevated WSG synthesis in S mutans , which results in dysregulated cariogenicity in vivo. Furthermore, structural analysis provides new insights regarding the lack of mannose monosaccharides, especially in the WSG synthesis of the SmudexA mutants. The biofilm phenotypes that are associated with the reduced glucose monosaccharide composition in both WSG and water-insoluble glucan shift the dental biofilm to reduce the cariogenic incidence of the SmudexA mutants. Taken together, these data reveal that EPS synthesis fine-tuning by the dexA gene results in a densely packed EPS matrix that may impede the glucose metabolism of WSG, thereby leading to the lack of an energy source for the bacteria. These results highlight dexA targeting as a potentially effective tool in dental caries management.


Assuntos
Biofilmes/crescimento & desenvolvimento , Dextranase/genética , Glucanos/biossíntese , Streptococcus mutans/enzimologia , Streptococcus mutans/genética , Streptococcus mutans/metabolismo , Água/química , Animais , Cárie Dentária , Feminino , Regulação Bacteriana da Expressão Gênica , Técnicas de Inativação de Genes , Glucanos/química , Glucose/metabolismo , Glucosiltransferases/genética , Humanos , Concentração de Íons de Hidrogênio , Masculino , Manose/metabolismo , Modelos Animais , Mutação , Fenótipo , Ratos , Ratos Sprague-Dawley , Streptococcus mutans/crescimento & desenvolvimento , Transcriptoma , Virulência
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