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1.
Sci Rep ; 11(1): 23800, 2021 12 10.
Article in English | MEDLINE | ID: mdl-34893687

ABSTRACT

The aim of this study was to evaluate the antimicrobial efficacy of non-thermal atmospheric plasma (NTAP) against Streptococcus mutans biofilms. Resin discs were fabricated, wet-polished, UV sterilized, and immersed in water for monomer extraction (37 °C, 24 h). Biofilms of bioluminescent S. mutans strain JM10 was grown on resin discs in anaerobic conditions for (37 °C, 24 h). Discs were divided into seven groups: control (CON), 2% chlorhexidine (CHX), only argon gas 150 s (ARG) and four NTAP treatments (30 s, 90 s, 120 s, 150 s). NTAP was applied using a plasma jet device. After treatment, biofilms were analyzed through the counting of viable colonies (CFU), bioluminescence assay (BL), scanning electron microscopy (SEM), and polymerase chain reaction (PCR). All NTAP-treated biofilm yielded a significant CFU reduction when compared to ARG and CON. BL values showed that NTAP treatment for 90 s, 120 s or 150 s resulted in statistically significantly lower metabolic activity when compared to the other groups. CHX displayed the lowest means of CFU and BL. SEM showed significant morphological changes in NTAP-treated biofilm. PCR indicated damage to the DNA structure after NTAP treatment. NTAP treatment was effective in lowering the viability and metabolism of S. mutans in a time-dependent manner, suggesting its use as an intraoral surface-decontamination strategy.


Subject(s)
Anti-Bacterial Agents/pharmacology , Biofilms/drug effects , Composite Resins , Plasma Gases/pharmacology , Streptococcus mutans/drug effects , Luminescent Measurements/methods , Microbial Sensitivity Tests , Microbial Viability/drug effects , Streptococcus mutans/ultrastructure , Surface Properties , Time Factors
2.
Sci Rep ; 11(1): 18290, 2021 09 14.
Article in English | MEDLINE | ID: mdl-34521969

ABSTRACT

Although fluoride has been widely used as a preventive agent for dental caries, the effects of fluoride on the activities of biofilms in different stages of cariogenic biofilm formation are less studied. This study was designed to investigate the antibiofilm activity of sodium fluoride during the early and mature stages of Streptococcus mutans (S. mutans) biofilm formation. S. mutans biofilms were formed on saliva-coated hydroxyapatite disks. In the early (0-46 h) and mature (46-94 h) biofilm stages, the biofilms were treated with different concentrations of fluoride (250, 500, 1000, 2000 ppm; 5 times in total, 1 min/treatment). Acidogenicity, dry weight, colony-forming units (CFUs), water-soluble/insoluble extracellular polysaccharides (EPSs), and intracellular polysaccharides were analysed, and confocal laser scanning microscopy images were obtained of the two stages of biofilms to determine antibiofilm activities of fluoride at varying concentrations during the formation of early and mature biofilms. In the early stages of cariogenic biofilm formation, test groups with all fluoride concentrations significantly inhibited the growth of S. mutans biofilms. The antibiofilm and anti-EPS formation activities of the brief fluoride treatments increased with a concentration-dependent pattern. At the mature biofilm stage, only the 2000 ppm fluoride treatment group significantly inhibited biofilm accumulation, activity, and intracellular/extracellular polysaccharide content compared with those of the control and other fluoride treatment groups. The antimicrobial effect of fluoride treatment on the growth of S. mutans biofilms was linked with the stage of cariogenic biofilm formation. The inhibition of S. mutans biofilm growth by fluoride treatment was easier in the early formation stage than in the mature stage. Fluoride treatment in the early stage of cariogenic biofilm formation may be an effective approach to controlling cariogenic biofilm development and preventing dental caries.


Subject(s)
Biofilms/drug effects , Sodium Fluoride/pharmacology , Streptococcus mutans/drug effects , Adult , Biofilms/growth & development , Dose-Response Relationship, Drug , Humans , Male , Microscopy, Confocal , Sodium Fluoride/administration & dosage , Streptococcus mutans/growth & development , Streptococcus mutans/ultrastructure
3.
Commun Biol ; 4(1): 846, 2021 07 15.
Article in English | MEDLINE | ID: mdl-34267305

ABSTRACT

Dental plaques are biofilms that cause dental caries by demineralization with acidogenic bacteria. These bacteria reside inside a protective sheath which makes any curative treatment challenging. We propose an antibiotic-free strategy to disrupt the biofilm by engineered clustered carbon dot nanoparticles that function in the acidic environment of the biofilms. In vitro and ex vivo studies on the mature biofilms of Streptococcus mutans revealed >90% biofilm inhibition associated with the contact-mediated interaction of nanoparticles with the bacterial membrane, excessive reactive oxygen species generation, and DNA fragmentation. An in vivo examination showed that these nanoparticles could effectively suppress the growth of S. mutans. Importantly, 16S rRNA analysis of the dental microbiota showed that the diversity and richness of bacterial species did not substantially change with nanoparticle treatment. Overall, this study presents a safe and effective approach to decrease the dental biofilm formation without disrupting the ecological balance of the oral cavity.


Subject(s)
Biofilms/drug effects , Microbiota/physiology , Nanoparticles/toxicity , Polymers/toxicity , Streptococcus mutans/drug effects , Animals , Anti-Bacterial Agents/pharmacology , Biofilms/growth & development , Female , Humans , Mice , Microbial Viability/drug effects , Microbiota/genetics , Microscopy, Atomic Force , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , NIH 3T3 Cells , Nanoparticles/chemistry , Nanoparticles/ultrastructure , Polymers/chemistry , RNA, Ribosomal, 16S/genetics , Rats, Sprague-Dawley , Streptococcus mutans/growth & development , Streptococcus mutans/ultrastructure
4.
Int J Nanomedicine ; 16: 3041-3057, 2021.
Article in English | MEDLINE | ID: mdl-33948084

ABSTRACT

BACKGROUND: The dentin exposure always leads to dentin hypersensitivity and/or caries. Given the dentin's tubular structure and low mineralization degree, reestablishing an effective biobarrier to stably protect dentin remains significantly challenging. This study reports a versatile dentin surface biobarrier consisting of a mesoporous silica-based epigallocatechin-3-gallate (EGCG)/nanohydroxyapatite delivery system and evaluates its stability on the dentinal tubule occlusion and the Streptococcus mutans (S. mutans) biofilm inhibition. MATERIALS AND METHODS: The mesoporous delivery system was fabricated and characterized. Sensitive dentin discs were prepared and randomly allocated to three groups: 1, control group; 2, casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) group; and 3, the mesoporous delivery system group. The dentin permeability, dentinal tubule occlusion, acid and abrasion resistance, and S. mutans biofilm inhibition were determined for 1 week and 1 month. The in vitro release profiles of EGCG, Ca, and P were also monitored. RESULTS: The mesoporous delivery system held the ability to sustainably release EGCG, Ca, and P and could persistently occlude dentinal tubules with acid and abrasion resistance, reduce the dentin permeability, and inhibit the S. mutans biofilm formation for up to 1 month compared with the two other groups. The system provided prolonged stability to combat oral adverse challenges and served as an effective surface biobarrier to protect the exposed dentin. CONCLUSION: The establishment of the dentin surface biobarrier consisting of a mesoporous delivery system indicates a promising strategy for the prevention and the management of dentin hypersensitivity and caries after enamel loss.


Subject(s)
Biofilms/growth & development , Dentin/chemistry , Streptococcus mutans/physiology , Acids , Adsorption , Biofilms/drug effects , Calcium/analysis , Caseins/pharmacology , Catechin/analogs & derivatives , Catechin/chemistry , Catechin/pharmacology , Cell Death/drug effects , Colony Count, Microbial , Dental Pulp/cytology , Humans , Nanoparticles/chemistry , Nanoparticles/ultrastructure , Nitrogen/chemistry , Permeability , Phosphorus/analysis , Porosity , Silicon Dioxide/chemistry , Streptococcus mutans/ultrastructure
5.
Biomed Res Int ; 2021: 6692328, 2021.
Article in English | MEDLINE | ID: mdl-33860052

ABSTRACT

To investigate the effects of ficin on biofilm formation of conditionally cariogenic Streptococcus mutans (S. mutans). Biomass and metabolic activity of biofilm were assessed using crystal violet assay, colony-forming unit (CFU) counting, and MTT assay. Extracellular polysaccharide (EPS) synthesis was displayed by SEM imaging, bacteria/EPS staining, and anthrone method while acid production was revealed by lactic acid assay. Growth curve and live/dead bacterial staining were conducted to monitor bacterial growth state in both planktonic and biofilm form. Total protein and extracellular proteins of S. mutans biofilm were analyzed by protein/bacterial staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), severally. qRT-PCR was conducted to detect acid production, acid tolerance, and biofilm formation associated genes. Crystal violet assay, CFU counting, and MTT assay showed that the suppression effect of ficin on S. mutans biofilm formation was concentration dependent. 4 mg/mL ficin had significant inhibitory effect on S. mutans biofilm formation including biomass, metabolic activity, EPS synthesis, and lactic acid production (p < 0.05). The growth curves from 0 mg/mL to 4 mg/mL ficin were aligned with each other. There was no significant difference among different ficin groups in terms of live/dead bacterial staining result (p > 0.05). Protein/bacterial staining outcome indicated that ficin inhibit both total protein and biofilm formation during the biofilm development. There were more relatively small molecular weight protein bands in extracellular proteins of 4 mg/mL ficin group when compared with the control. Generally, ficin could inhibit biofilm formation and reduce cariogenic virulence of S. mutans effectively in vitro; thus, it could be a potential anticaries agent.


Subject(s)
Biofilms/growth & development , Ficain/pharmacology , Streptococcus mutans/physiology , Biomass , Colony Count, Microbial , Gene Expression Regulation, Bacterial/drug effects , Lactic Acid/biosynthesis , Microbial Viability/drug effects , Polysaccharides, Bacterial/biosynthesis , RNA, Messenger/genetics , RNA, Messenger/metabolism , Streptococcus mutans/drug effects , Streptococcus mutans/metabolism , Streptococcus mutans/ultrastructure
6.
Microb Pathog ; 143: 104129, 2020 Jun.
Article in English | MEDLINE | ID: mdl-32169491

ABSTRACT

Dental caries is a common cause for tooth loss and Streptococcus mutans is identified as the etiologic pathogen. This study evaluates the inhibitory potential of Epigallocatechin gallate (EGCG) on S.mutans glucansucrase enzyme and its biofilm. Glucansucrase binding and the inhibitory potential of EGCG was validated using AutoDock tool and enzyme inhibitory assay. Biofilm inhibitory potential was also confirmed using Scanning Electron Microscopic (SEM) analysis in human tooth samples. Molecular docking revealed that EGCG interacted with GLU 515 and TRP 517 amino acids and binds to glucansucrase. SEM analysis revealed inhibition of S.mutans biofilm by various concentrations of EGCG on surfaces of tooth samples. Bioinformatics and biological assays confirmed that EGCG potentially binds to the S. mutans glucansucrase and inhibits its enzymatic activity. Enzymatic inhibition of glucansucrase attenuated biofilm formation potential of S. mutans on tooth surface. Thus, we conclude that EGCG inhibitory potential of S. mutans biofilm on the tooth surface is a novel approach in prevention of dental caries.


Subject(s)
Biofilms/drug effects , Catechin/analogs & derivatives , Dental Caries/prevention & control , Streptococcus mutans/drug effects , Catechin/pharmacology , Catechin/therapeutic use , Dental Caries/microbiology , Humans , Microscopy, Electron, Scanning , Molecular Docking Simulation , Streptococcus mutans/ultrastructure , Tooth/microbiology
7.
Lasers Med Sci ; 35(3): 709-718, 2020 Apr.
Article in English | MEDLINE | ID: mdl-31713778

ABSTRACT

The potential anti-cariogenic effect of blue light was evaluated using an oral biofilm model. Two species, Streptococcus mutans and Streptococcus sanguinis, were cultivated ex vivo on bovine enamel blocks for 24 h, either separately or mixed together, then exposed to blue light (wavelengths 400-500 nm) using 112 J/cm2. Twenty four or 48 h after exposure to light the biofilm structure and biomass were characterized and quantified using SEM and qPCR, respectively. Bacterial viability was analyzed by CLSM using live/dead bacterial staining. Gene expression was examined by RT-qPCR. After exposure to light, S. mutans biomass in mono-species biofilm was increased mainly by dead bacteria, relative to control. However, the bacterial biomass of S. mutans when grown in mixed biofilm and of S. sanguinis in mono-species biofilm was reduced after light exposure, with no significant change in viability when compared to control. Furthermore, when grown separately, an upregulation of gene expression related to biofilm formation of S. mutans, and downregulation of similar genes of S. sanguinis, were measured 24 h after exposure to blue light. However, in mixed biofilm, a downregulation of those genes in both species was observed, although not significant in S. mutans. In conclusion, blue light seems to effectively alter the bacterial biomass by reducing the viability and virulence characteristics in both bacterial species and may promote the anti-cariogenic balance between them, when grown in a mixed biofilm. Therefore, exposure of oral biofilm to blue light has the potential to serve as a complementary approach in preventive dentistry.


Subject(s)
Biofilms/radiation effects , Light , Models, Biological , Mouth/microbiology , Streptococcus mutans/radiation effects , Streptococcus sanguis/radiation effects , Animals , Biofilms/growth & development , Cattle , Dental Enamel/microbiology , Dental Enamel/ultrastructure , Gene Expression Regulation, Bacterial/radiation effects , Streptococcus mutans/genetics , Streptococcus mutans/ultrastructure , Streptococcus sanguis/genetics , Streptococcus sanguis/ultrastructure
8.
Biomed Res Int ; 2019: 1950790, 2019.
Article in English | MEDLINE | ID: mdl-31781595

ABSTRACT

The present study aimed at investigating the influence of norspermidine on the formation of dual-species biofilms composed of Streptococcus mutans (S. mutans) and Streptococcus sanguinis (S. sanguinis). Crystal violet assay was conducted to assess the formation of single-species biofilms of S. mutans and S. sanguinis, and the growth curve was carefully observed to monitor the growth of these two species of bacteria. Fluorescence in situ hybridization (FISH) and MTT array were used to analyze the composition and metabolic activity of the dual-species biofilms, respectively. Extracellular polysaccharides (EPS)/bacteria staining, anthrone method, and scanning electron microscopy (SEM) imaging were conducted to study the synthesis of EPS by dual-species biofilms. Lactic acid assay and pH were measured to detect dual-species biofilm acid production. We found that norspermidine had different effects on S. mutans and S. sanguinis including their growth and biofilm formation. Norspermidine regulated the composition of the dual-species biofilms, decreased the ratio of S. mutans in dual-species biofilms, and reduced the metabolic activity, EPS synthesis, and acid production of dual-species biofilms. Norspermidine regulated dual-species biofilms in an ecological way, suggesting that it may be a potent reagent for controlling dental biofilms and managing dental caries.


Subject(s)
Biofilms/drug effects , Cariogenic Agents/pharmacology , Dental Caries/prevention & control , Polysaccharides, Bacterial/metabolism , Spermidine/analogs & derivatives , Streptococcus mutans/drug effects , Streptococcus sanguis/drug effects , Biofilms/growth & development , Dental Caries/drug therapy , Dental Caries/microbiology , Hydrogen-Ion Concentration , In Situ Hybridization, Fluorescence , Microbial Interactions , Microscopy, Electron, Scanning , Spermidine/pharmacology , Streptococcus mutans/growth & development , Streptococcus mutans/ultrastructure , Streptococcus sanguis/growth & development , Streptococcus sanguis/ultrastructure , Virulence
9.
ACS Appl Mater Interfaces ; 11(46): 42943-42955, 2019 Nov 20.
Article in English | MEDLINE | ID: mdl-31647216

ABSTRACT

The capability to detect bacteria at a low cell density is critical to prevent the delay in therapeutic intervention and to avoid the emergence of antibiotic-resistant species. Till date, significant advancement has been made to develop a sensing platform for rapid and reliable bacterial detection. However, critical requirements, that is, limit of detection, fast time of response, ultrasensitivity with high reproducibility, and the ability to distinguish between bacterial strains are yet to be met within a single sensing platform. In this contribution, we present a novel label-free sensor based on pH-sensitive fluorescent yttrium-doped carbon nanoparticles (YCNPs) embedded in agarose that can rapidly and accurately detect and discriminate pathogens in real time. The developed sensor matrix presented pH-triggered aggregation-induced emission quenching of YCNPs in a wide pH range. When the pH decreased from 10.0 to 4.0, the fluorescence of the matrix decreased linearly (R2 = 0.9229). The sensor 's high sensitivity in a physiologically relevant pH range enables the monitoring of the presence of live pathogens to single-cell resolution. In addition, the 3D matrix sensor showed low cytotoxicity and long stability (>30 days). Besides, the YCNP platform is stable for several hours (5 h) in a complex medium and does not alter the bacterial activities, allowing real-time monitoring of bacterial growth with a small sample volume (100 µL) and rapid response time (25 min). Furthermore, using machine learning-assisted tools, different bacterial strains with various cell densities were discriminated with an accuracy of almost 100%. Moreover, blends of pathogens and a real-world sample can also be identified accurately, thus enabling the sensor to provide fast and reliable pathogen information for clinical decisions and allowing continuous monitoring of infectious disease trends.


Subject(s)
Carbon/chemistry , Nanoparticles/chemistry , Streptococcus mutans , Yttrium/chemistry , Hydrogen-Ion Concentration , Limit of Detection , Nanoparticles/ultrastructure , Streptococcus mutans/metabolism , Streptococcus mutans/ultrastructure
10.
Microbiologyopen ; 8(4): e00695, 2019 04.
Article in English | MEDLINE | ID: mdl-30051653

ABSTRACT

For caries-active patients, antimicrobial measures may be useful in addition to mechanical biofilm removal. The aim of this study was to investigate the antimicrobial efficacy of alternative compounds for use in oral care from two main categories (i.e., preservatives and natural compounds) toward biofilms from caries-associated bacteria as compared to oral care gold-standards chlorhexidine digluconate (CHX), cetylpyridinium chloride (CPC), and zinc. Compounds were screened in initial Streptococcus mutans biofilms. Then, the most effective compounds were further investigated in mature S. mutans and polymicrobial biofilms comprising Actinomyces naeslundii, Actinomyces odontolyticus, and S. mutans. Here, distinct treatment periods and concentrations were evaluated. Biofilms were visualized by scanning electron microscopy and bacterial membrane damage was evaluated by means of flow cytometry and staining with SYBR Green and propidium iodide. Citrus extract was the only compound exhibiting similar antimicrobial efficacy in initial S. mutans biofilms (>5 log10 ) as compared to CHX and CPC, but its effect was clearly inferior in mature S. mutans and polymicrobial biofilms. Flow cytometric data suggested that the mechanism of antimicrobial action of citrus extract may be based on damage of bacterial membranes similar to CHX and CPC. From all alternative compounds investigated in this study, citrus extract exhibited the highest antimicrobial efficacy toward in vitro biofilms from caries-associated bacteria, but still was less effective than oral care gold-standard antiseptics CHX and CPC. Nevertheless, citrus extract may be a valuable antimicrobial compound for use in oral care for caries-active patients.


Subject(s)
Anti-Bacterial Agents/pharmacology , Anti-Infective Agents, Local/pharmacology , Biofilms , Dental Caries/microbiology , Streptococcal Infections/microbiology , Streptococcus mutans/drug effects , Cetylpyridinium/pharmacology , Chlorhexidine/analogs & derivatives , Chlorhexidine/pharmacology , Humans , Microbial Sensitivity Tests , Microscopy, Electron, Scanning , Streptococcus mutans/physiology , Streptococcus mutans/ultrastructure
11.
Int J Nanomedicine ; 13: 7697-7709, 2018.
Article in English | MEDLINE | ID: mdl-30538453

ABSTRACT

BACKGROUND: Oral plaque biofilms pose a threat to periodontal health and are challenging to eradicate. There is a growing belief that a combination of silver nanoparticles and chlorhexidine (CHX) is a promising strategy against oral biofilms. PURPOSE: To overcome the side effects of this strategy and to exert maximum efficiency, we fabricated biodegradable disulfide-bridged mesoporous silica nanoparticles (MSNs) to co-deliver silver nanoparticles and CHX for biofilm inhibition. MATERIALS AND METHODS: CHX-loaded, silver-decorated mesoporous silica nanoparticles (Ag-MSNs@CHX) were fabricated after CHX loading, and the pH- and glutathione-responsive release profiles of CHX and silver ions along with their mechanism of degradation were systematically investigated. Then, the efficacy of Ag-MSNs@CHX against Streptococcus mutans and its biofilm was comprehensively assessed by determining the minimum inhibitory concentration, minimum bactericidal concentration, minimal biofilm inhibitory concentration, and the inhibitory effect on S. mutans biofilm formation. In addition, the biosafety of nanocarriers was evaluated by oral epithelial cells and a mouse model. RESULTS: The obtained Ag-MSNs@CHX possessed redox/pH-responsive release properties of CHX and silver ions, which may be attributed to the redox-triggered matrix degradation mechanism of exposure to biofilm-mimetic microenvironments. Ag-MSNs@CHX displayed dose-dependent antibacterial activity against planktonic and clone formation of S. mutans. Importantly, Ag-MSNs@CHX had an increased and long-term ability to restrict the growth of S. mutans biofilms compared to free CHX. Moreover, Ag-MSNs@CHX showed less cytotoxicity to oral epithelial cells, whereas orally administered Ag-MSNs exhibited no obvious toxic effects in mice. CONCLUSION: Our findings constitute a highly effective and safe strategy against biofilms that has a good potential as an oral biofilm therapy.


Subject(s)
Biocompatible Materials/chemistry , Biofilms/drug effects , Chlorhexidine/pharmacology , Metal Nanoparticles/chemistry , Mouth/microbiology , Silicon Dioxide/chemistry , Silver/pharmacology , Animals , Anti-Bacterial Agents/pharmacology , Cell Death/drug effects , Delayed-Action Preparations/pharmacology , Drug Liberation , Epithelial Cells/drug effects , Epithelial Cells/pathology , Humans , Hydrogen-Ion Concentration , Ions , Metal Nanoparticles/ultrastructure , Mice , Microbial Sensitivity Tests , Nanoparticles/chemistry , Organ Specificity , Oxidation-Reduction , Streptococcus mutans/drug effects , Streptococcus mutans/ultrastructure
12.
Sci Rep ; 8(1): 15818, 2018 10 25.
Article in English | MEDLINE | ID: mdl-30361619

ABSTRACT

Soft lithography and Dip-Pen Nanolithography (DPN) are techniques that have been used to modify the surface of biomaterials. Modified surfaces play a role in reducing bacterial adhesion and biofilm formation. Also, titanium dioxide has been reported as an antibacterial substance due to its photocatalytic effect. This work aimed at creating patterns on model surfaces using DPN and soft lithography combined with titanium dioxide to create functional antibacterial micropatterned surfaces, which were tested against Streptococcus mutans. DPN was used to create a master pattern onto a model surface and microstamping was performed to duplicate and transfer such patterns to medical-grade stainless steel 316L using a suspension of TiO2. Modified SS316L plates were subjected to UVA black light as photocatalytic activator. Patterns were characterized by atomic force microscopy and biologically evaluated using S. mutans. A significant reduction of up to 60% in bacterial adhesion to TiO2 -coated and -micropatterned surfaces was observed. Moreover, both TiO2 surfaces reduced the viability of adhered bacteria after UV exposure. TiO2 micropatterned demonstrated a synergic effect between physical and chemical modification against S. mutans. This dual effect was enhanced by increasing TiO2 concentration. This novel approach may be a promising alternative to reduce bacterial adhesion to surfaces.


Subject(s)
Anti-Bacterial Agents/pharmacology , Nanotechnology/methods , Printing , Titanium/pharmacology , Bacterial Adhesion/drug effects , Coated Materials, Biocompatible/pharmacology , Dimethylpolysiloxanes/chemistry , Microbial Viability/drug effects , Microscopy, Atomic Force , Spectrometry, X-Ray Emission , Streptococcus mutans/drug effects , Streptococcus mutans/ultrastructure , Surface Properties , Water/chemistry
13.
Int J Nanomedicine ; 13: 3751-3762, 2018.
Article in English | MEDLINE | ID: mdl-29988768

ABSTRACT

INTRODUCTION: The placement of dental implants is performed in a contaminated surgical field in the oral cavity, which may lead to implant failure. Bacterial adhesion and proliferation (Streptococcus mutans, Porphyromonas gingivalis) often lead to implant infections. Although Ag nanoparticles hold great promise for a broad spectrum of antibacterial activities, their runoff from dental implants compromises their antibacterial efficacy and potentially impairs osteoblast proliferation. Thus, this aspect remains a primary challenge and should be controlled. MATERIALS AND METHODS: In this study, PLGA(Ag-Fe3O4) was modified on the implanted tooth surface and was characterized by transmission electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy. The magnetic and antibacterial properties were also determined. RESULTS: Results showed that Ag successfully bonded with Fe3O4, and Ag-Fe3O4 not only exerted superparamagnetism but also exhibited antibacterial activity almost identical to silver nanoparticles (nano-Ag). The PLGA(Ag-Fe3O4) coating could significantly maintain the antibacterial activity and avoid bacterial adhesion to the implant. Compared with the blank control group, PLGA(Ag-Fe3O4) under magnetic field-coated samples had a significantly lower amount of colonized S. mutans (P<0.01). Osteoblast proliferation results showed that the coated samples did not exhibit cytotoxicity and could promote osteoblast proliferation as shown by MTT, alkaline phosphatase, and the nucleolar organizer region count. CONCLUSION: We developed a novel Ag biologically compatible nanoparticle in this study without compromising the nano-Ag antibacterial activity, which provided continuous antibacterial action.


Subject(s)
Bacterial Adhesion/drug effects , Coated Materials, Biocompatible/pharmacology , Dental Implants/microbiology , Ferric Compounds/pharmacology , Lactic Acid/chemistry , Magnetic Fields , Osteogenesis/drug effects , Polyglycolic Acid/chemistry , Silver/pharmacology , Animals , Anti-Bacterial Agents/pharmacology , Cell Proliferation/drug effects , Female , Male , Metal Nanoparticles/chemistry , Metal Nanoparticles/ultrastructure , Mice , Microbial Sensitivity Tests , Nanocomposites/chemistry , Nanocomposites/ultrastructure , Polylactic Acid-Polyglycolic Acid Copolymer , Porphyromonas gingivalis/drug effects , Rats, Sprague-Dawley , Streptococcus mutans/drug effects , Streptococcus mutans/ultrastructure
14.
Drug Discov Ther ; 12(3): 133-141, 2018.
Article in English | MEDLINE | ID: mdl-29998994

ABSTRACT

The overgrowth of certain strains of normal flora in oral cavity can cause many kinds of oral infections or diseases such as carries, periodontitis, and gingivitis. Prevention and treatment of these diseases are usually achieved by chemical antiseptics. However, these chemicals are found as negative impacts of human health hazards and accession of microbial resistance. The present study explores the potential of Piper betle extracts on inhibition of two oral pathogenic bacteria; Streptococcus mutans DMST 41283 and Streptococcus intermedius DMST 42700. P. betle demonstrated significantly higher inhibitory activity against both pathogenic strains than Acacia catechu, Camellia sinensis, Coccinia grandis, Solanum indicum, and Streblus asper. Among fractionated extracts of P. betle from several solvents, the extract from ethyl acetate (Pb-EtOAc) possessed the widest inhibition zone of 11.0 ± 0.1 and 11.3 ± 0.4 mm against both bacterial strains, respectively. Pb-EtOAc showed the same minimum inhibitory concentration of 0.5 mg/mL against both strains, whereas its minimum bactericidal concentrations were 2.0 and 0.5 mg/mL against S. mutans and S. intermedius, respectively. HPLC analysis demonstrated that the major active compound of Pb-EtOAc was 4-allylpyrocatechol. It was found that the killing kinetics of Pb-EtOAc against both test strains were time and dose dependent. Scanning electron microscopy micrographs showed the morphological changes and depletion of the tested pathogens indicating cell destruction after exposure to Pb-EtOAc. It is confirmed that Pb-EtOAc is potentially effective against both oral pathogens and might be used as natural alternative agents in prevention and treatment of oral infections caused by oral pathogenic bacteria.


Subject(s)
Piper betle , Plant Extracts/pharmacology , Streptococcus intermedius/drug effects , Streptococcus mutans/drug effects , Acacia , Camellia sinensis , Catechols/chemistry , Catechols/pharmacology , Chromatography, High Pressure Liquid , Cucurbitaceae , Dental Caries/microbiology , Gingivitis/microbiology , Humans , Microbial Sensitivity Tests , Microscopy, Electron, Scanning , Moraceae , Periodontitis/microbiology , Piper betle/chemistry , Plant Extracts/chemistry , Solanum , Streptococcus intermedius/ultrastructure , Streptococcus mutans/ultrastructure
15.
Arch Oral Biol ; 87: 226-234, 2018 Mar.
Article in English | MEDLINE | ID: mdl-29328950

ABSTRACT

OBJECTIVE: To study the ultrastructural alterations induced in Streptococcus mutans (ATCC 25175) incubated with saliva, saliva plus histatin 5 and histatin 5. METHODS: S. mutans incubated with saliva histatin 5 or a combination of both were morphologically analyzed and counted. The results were expressed as (CFU)ml-1. Ultrastructural damage was evaluated by transmission electron microscopy. Ultrastructural localization of histatin 5 was examined using immunogold labeling. Apoptotic cell death was determined by flow cytometry (TUNEL). RESULTS: A decrease in the bacteria numbers was observed after incubation with saliva, saliva with histatin 5 or histatin 5 compared to the control group (p<0.0001). Ultrastructural damage in S. mutans incubated with saliva was found in the cell wall. Saliva plus histatin 5 induced a cytoplasmic granular pattern and decreased the distance between the plasma membrane bilayers, also found after incubation with histatin 5, together with pyknotic nucleoids. Histatin 5 was localized on the bacterial cell walls, plasma membranes, cytoplasm and nucleoids. Apoptosis was found in the bacteria incubated with saliva (63.9%), saliva plus histatin 5 (71.4%) and histatin 5 (29.3%). Apoptosis in the control bacteria was 0.2%. CONCLUSIONS: Antibacterial activity against S. mutans and the morphological description of damage induced by saliva and histatin 5 was demonstrated. Pyknotic nucleoids observed in S. mutans exposed to saliva, saliva plus histatin 5 and histatin 5 could be an apoptosis-like death mechanism. The knowledge of the damage generated by histatin 5 and its intracellular localization could favor the design of an ideal peptide as a therapeutic agent.


Subject(s)
Histatins/pharmacology , Saliva/chemistry , Streptococcus mutans/drug effects , Streptococcus mutans/ultrastructure , Apoptosis , Cell Membrane/drug effects , Cell Membrane/ultrastructure , Cell Wall/drug effects , Cell Wall/ultrastructure , In Situ Nick-End Labeling , Microscopy, Electron, Transmission
16.
Nan Fang Yi Ke Da Xue Xue Bao ; 37(6): 786-791, 2017 Jun 20.
Article in Chinese | MEDLINE | ID: mdl-28669953

ABSTRACT

OBJECTIVE: To construct a SMU.2055-dificient mutant strain of Streptococcus mutans (S. mutans) and evaluate its cariogenic capacity in comparison with wild-type S. mutans. METHODS: The SMU.2055-dificient mutant strain of S. mutans was constructed using homologous recombination technique and observed with scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The absorbance at 600 nm and pH values of the wild-type and mutant strains were monitored to evaluate their growth and acid production. After acid adaption, the two strains were challenged with acid shock and their survival rates were determined. RESULTS: PCR and sequence analyses verified the successful construction of the SMU.2055-dificient mutant strain. Observation with SEM revealed obvious changes in the morphology of the mutant strain, which showed reduced irregular substances between the individual bacteria as compared with the wild-type strain. TEM revealed major alterations in the cellular architecture of the mutant strain with blurry cell membrane and disruption of the membrane integrity. The growth capacity of the mutant strain decreased in both normal and acidic conditions but its acid production capacity remained unaffected. CONCLUSION: SMU.2055 gene is associated with morphology maintenance, growth capacity and acid resistance of S. mutans but is not related to the acid production capacity of the bacterium.


Subject(s)
Bacterial Proteins/genetics , Streptococcus mutans/genetics , Streptococcus mutans/ultrastructure , Acids/metabolism , Dental Caries/microbiology , Genes, Bacterial , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Streptococcus mutans/pathogenicity
17.
J Biomed Mater Res A ; 105(9): 2632-2639, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28512783

ABSTRACT

Orthodontic retention has been proposed as a life-long commitment for patients who desire to maintain straight teeth. However, the presence of foreign material increases risk of bacterial colonization and caries formation, of which Streptococcus mutans is a key contributor. Multiple studies have assessed the ability of silver to be added to base plate material and resist attachment of S. mutans. However, it does not appear that long-term washout in connection with biofilm growth under physiologically relevant conditions has been taken into consideration. In this study, silver was added to base plate material and exposed to short- or long-term washout periods. Materials were then assessed for their ability to resist biofilm formation of S. mutans using a drip flow reactor that modeled the human oral environment. Data indicated that silver was able to resist biofilm formation following short-term washout, but long-term washout periods resulted in a lack of ability to resist biofilm formation. These data will be important for future development of base plate materials to achieve long-term antimicrobial efficacy to reduce risk of caries formation and benefit patients in the long term. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2632-2639, 2017.


Subject(s)
Bioreactors/microbiology , Polymethyl Methacrylate/pharmacology , Silver/pharmacology , Streptococcus mutans/drug effects , Biofilms/drug effects , Biofilms/growth & development , Imaging, Three-Dimensional , Microbial Sensitivity Tests , Streptococcus mutans/ultrastructure , Surface Properties
18.
J Mol Recognit ; 30(7)2017 07.
Article in English | MEDLINE | ID: mdl-28256775

ABSTRACT

Streptococcus mutans cells form robust biofilms on human teeth and are strongly related to caries incidents. Hence, understanding the adhesion of S. mutans in the human oral cavity is of major interest for preventive dentistry. In this study, we report on atomic force microscopy-based single-cell force spectroscopy measurements of S. mutans cells to hydroxyapatite surfaces. We observe for almost all measurements a significant difference in adhesion strength for S. mutans as well as for Staphylococcus carnosus cells. However, the increase in adhesion strength after saliva exposure is much higher for S. mutans cells compared to S. carnosus cells. Our results demonstrate that S. mutans cells are well adapted to their natural environment, the oral cavity. This ability promotes the biofilm-forming capability of that species and hence the production of caries-provoking acids. In consequence, understanding the fundamentals of this mechanism may pave a way towards more effective caries-reducing techniques.


Subject(s)
Biofilms/drug effects , Cell Adhesion/drug effects , Saliva/chemistry , Streptococcus mutans/drug effects , Biofilms/growth & development , Durapatite/chemistry , Humans , Microscopy, Atomic Force , Saliva/microbiology , Single-Cell Analysis , Streptococcus mutans/pathogenicity , Streptococcus mutans/ultrastructure , Tooth/microbiology , Tooth/ultrastructure
19.
PLoS One ; 12(3): e0174440, 2017.
Article in English | MEDLINE | ID: mdl-28350880

ABSTRACT

A comparative study of the corrosion resistance of CoCr and NiCr alloys in artificial saliva (AS) containing tryptic soy broth (Solution 1) and Streptococcus mutans (S. mutans) species (Solution 2) was performed by electrochemical methods, including open circuit potential measurements, impedance spectroscopy, and potentiodynamic polarization. The adherence of S. mutans to the NiCr and CoCr alloy surfaces immersed in Solution 2 for 24 h was verified by scanning electron microscopy, while the results of electrochemical impedance spectroscopy confirmed the importance of biofilm formation for the corrosion process. The R(QR) equivalent circuit was successfully used to fit the data obtained for the AS mixture without S. mutans, while the R(Q(R(QR))) circuit was found to be more suitable for describing the biofilm properties after treatment with the AS containing S. mutans species. In addition, a negative shift of the open circuit potential with immersion time was observed for all samples regardless of the solution type. Both alloys exhibited higher charge transfer resistance after treatment with Solution 2, and lower corrosion current densities were detected for all samples in the presence of S. mutans. The obtained results suggest that the biofilm formation observed after 24 h of exposure to S. mutans bacteria might enhance the corrosion resistance of the studied samples by creating physical barriers that prevented oxygen interactions with the metal surfaces.


Subject(s)
Corrosion , Dental Alloys/chemistry , Saliva, Artificial/pharmacology , Streptococcus mutans/drug effects , Bacterial Adhesion/drug effects , Biofilms/drug effects , Biofilms/growth & development , Chromium Alloys/chemistry , Chromium Alloys/standards , Cobalt/chemistry , Dental Alloys/standards , Dielectric Spectroscopy , Electrochemical Techniques , Humans , Hydrogen-Ion Concentration , Materials Testing/methods , Microscopy, Electron, Scanning , Nickel/chemistry , Saliva, Artificial/chemistry , Streptococcus mutans/physiology , Streptococcus mutans/ultrastructure , Surface Properties , Time Factors
20.
Sci Rep ; 7: 41787, 2017 02 07.
Article in English | MEDLINE | ID: mdl-28169312

ABSTRACT

Irreversible white spot lesion (WSL) occurs in up to 50% of patients during orthodontic treatment. Therefore, orthodontic adhesives need to be able to inhibit or reduce bacterial growth in order to prevent or minimize WSL. This study evaluated the antibacterial effect and shear bond strength (SBS) of a resin-based orthodontic adhesive containing the antibacterial monomer 2-methacryloxylethyl hexadecyl methyl ammonium bromide (MAE-HB). MAE-HB was added at three concentrations (1, 3, and 5 wt%) to a commercial orthodontic adhesive Transbond XT, while the blank control comprised unmodified Transbond XT. Their antibacterial effects on Streptococcus mutans were investigated after 0 and 180 days of aging. The SBS of metal brackets bonded to the buccal enamel surface of human premolars was assessed. Compared with the blank control, the MAE-HB-incorporated adhesive exhibited a significant contact inhibitory effect on the growth of S. mutans (P < 0.05), even after 180 days of aging. SBS and adhesive remnant index values revealed that the bonding ability of the experimental adhesive was not significantly adversely affected by the incorporation of MAE-HB at any of the three concentrations. Therefore, orthodontic adhesives with strong and long-lasting bacteriostatic properties can be created through the incorporation of MAE-HB without negatively influencing bonding ability.


Subject(s)
Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Dental Cements/chemistry , Dental Cements/pharmacology , Methacrylates/chemistry , Methacrylates/pharmacology , Quaternary Ammonium Compounds/chemistry , Quaternary Ammonium Compounds/pharmacology , Biofilms/drug effects , Humans , Microbial Sensitivity Tests , Shear Strength , Streptococcus mutans/drug effects , Streptococcus mutans/growth & development , Streptococcus mutans/ultrastructure
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