scholarly journals Antibacterial Photodynamic Inactivation of Fagopyrin F from Tartary Buckwheat (Fagopyrum tataricum) Flower against Streptococcus mutans and Its Biofilm

2021 ◽  
Vol 22 (12) ◽  
pp. 6205
Author(s):  
Jaecheol Kim ◽  
Suna Kim ◽  
Kiuk Lee ◽  
Ryun Hee Kim ◽  
Keum Taek Hwang
Keyword(s):  
Streptococcus Mutans ◽  
Laser Scanning ◽  
Light Exposure ◽  
Laser Scanning Microscopy ◽  
Tartary Buckwheat ◽  
Plate Count ◽  
Confocal Laser ◽  
Light Sources ◽  
Photodynamic Inactivation ◽  
Fagopyrum Tataricum

The objective of this study was to determine reactive oxygen species (ROS) produced by fagopyrin F-rich fraction (FFF) separated from Tartary buckwheat flower extract exposed to lights and to investigate its antibacterial photodynamic inactivation (PDI) against Streptococcus mutans and its biofilm. ROS producing mechanisms involving FFF with light exposure were determined using a spectrophotometer and a fluorometer. S. mutans and its biofilm inactivation after PDI treatment of FFF using blue light (BL; 450 nm) were determined by plate count method and crystal violet assay, respectively. The biofilm destruction by ROS produced from FFF after exposure to BL was visualized using confocal laser scanning microscopy (CLSM) and field emission scanning electron microscope (FE-SEM). BL among 3 light sources produced type 1 ROS the most when applying FFF as a photosensitizer. FFF exposed to BL (5 and 10 J/cm2) significantly more inhibited S. mutans viability and biofilm formation than FFF without the light exposure (p < 0.05). In the PDI of FFF exposed to BL (10 J/cm2), an apparent destruction of S. mutans and its biofilm were observed by the CLSM and FE-SEM. Antibacterial PDI effect of FFF was determined for the first time in this study.

scholarly journals Effects of cigarette smoking on the growth of Streptococcus mutans biofilms: an in vitro study

2021 ◽  
Author(s):  
Ye Han
Keyword(s):  
Treatment Group ◽  
Cigarette Smoking ◽  
Streptococcus Mutans ◽  
Laser Scanning ◽  
Water Soluble ◽  
Laser Scanning Microscopy ◽  
Extracellular Polysaccharides ◽  
Confocal Laser ◽  
Control Group

Abstract This study aimed to investigate the differences in growth and virulence (EPSs and acidogenicity) of Streptococcus mutans biofilms (S. mutans) according to the different times of cigarette smoking (CS) treatment. S. mutans biofilms (74-hour-old) were formed on saliva-coated hydroxyapatite disks. The biofilms were treated with CS at different times per day (one time, three times, and six times/day). The control group did not receive CS treatment. Acidogenicity, dry weight, colony-forming units, water-soluble/insoluble extracellular polysaccharides, and intracellular polysaccharides were analyzed and confocal laser scanning microscopy images were obtained of the 74-h-old biofilms. The 74-h-old biofilms on sHA discs in the 6 times/day CS treatment group showed the lowest biofilm accumulation and extracellular polysaccharide amount compared with the control group and other CS treatment groups. In the CLSM study, the biofilms in the six times/day CS treatment group also showed the lowest bacterial count (live and dead cells) and EPS biovolume. CS has an obvious inhibition on the growth of S. mutans biofilms, the degree of inhibition is proportional to the number of CS treatments.

scholarly journals Assessment of Genes Associated with Streptococcus mutans Biofilm Morphology

2006 ◽  
Vol 72 (9) ◽  
pp. 6277-6287 ◽  
Author(s):  
Mizuho Motegi ◽  
Yuzo Takagi ◽  
Hideo Yonezawa ◽  
Nobuhiro Hanada ◽  
Jun Terajima ◽  
...  
Keyword(s):  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Protein Gene ◽  
Laser Scanning Microscopy ◽  
Live Cells ◽  
Confocal Laser ◽  
Morphological Development ◽  
Phosphotransferase System ◽  
Whole Saliva

ABSTRACT Streptococcus mutans, the major pathogen responsible for dental caries in humans, is a biofilm-forming bacterium. In the present study, 17 different pulsed-field gel electrophoresis patterns of genomic DNA were identified in S. mutans organisms isolated clinically from whole saliva. The S. mutans isolates showed different abilities to form biofilms on polystyrene surfaces in semidefined minimal medium cultures. Following cultivation in a flow cell system in tryptic soy broth with 0.25% sucrose and staining using a BacLight LIVE/DEAD system, two strains, designated FSC-3 and FSC-4, showed the greatest and least, respectively, levels of biofilm formation when examined with confocal laser scanning microscopy. Further, image analyses of spatial distribution and architecture were performed to quantify the merged green (live cells) and red (dead cells) light. The light intensity of the FSC-3 biofilm was greater than that of the FSC-4 biofilm in the bottom area but not in the top area. S. mutans whole-genome array results showed that approximately 3.8% of the genes were differentially expressed in the two strains, of which approximately 2.2%, including bacitracin transport ATP-binding protein gene glrA and a BLpL-like putative immunity protein gene, were activated in FSC-3. In addition, about 1.6% of the genes, including those associated with phosphotransferase system genes, were repressed. Analyses of the glrA-deficient strains and reverse transcription-PCR confirmed the role of the gene in biofilm formation. Differential assessment of biofilm-associated genes in clinical strains may provide useful information for understanding the morphological development of streptococcal biofilm, as well as for colonization of S. mutans.

Role of the luxS Gene in Initial Biofilm Formation by Streptococcus mutans

2015 ◽  
Vol 25 (1) ◽  
pp. 60-68 ◽  
Author(s):  
Zhiyan He ◽  
Jingping Liang ◽  
Zisheng Tang ◽  
Rui Ma ◽  
Huasong Peng ◽  
...  
Keyword(s):  
Streptococcus Mutans ◽  
Mutant Strain ◽  
Biofilm Formation ◽  
Type Strain ◽  
Laser Scanning ◽  
Wild Type Strain ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Wild Type ◽  
Initial Biofilm

Quorum sensing (QS) is a process by which bacteria communicate with each other by secreting chemical signals called autoinducers (AIs). Among Gram-negative and Gram-positive bacteria, AI-2 synthesized by the LuxS enzyme is widespread. The aim of this study was to evaluate the effect of QS <i>luxS</i> gene on initial biofilm formation by <i>Streptococcus mutans</i>. The bacterial cell surface properties, including cell hydrophobicity (bacterial adherence to hydrocarbons) and aggregation, which are important for initial adherence during biofilm development, were investigated. The biofilm adhesion assay was evaluated by the MTT method. The structures of the 5-hour biofilms were observed by using confocal laser scanning microscopy, and QS-related gene expressions were investigated by real-time PCR. The <i>luxS</i> mutant strain exhibited higher biofilm adherence and aggregation, but lower hydrophobicity than the wild-type strain. The confocal laser scanning microscopy images revealed that the wild-type strain tended to form smaller aggregates with uniform distribution, whereas the <i>luxS</i> mutant strain aggregated into distinct clusters easily discernible in the generated biofilm. Most of the genes examined were downregulated in the biofilms formed by the <i>luxS</i> mutant strain, except the <i>gtfB </i>gene. QS <i>luxS</i> gene can affect the initial biofilm formation by <i>S. mutans.</i>

scholarly journals The vicK gene of Streptococcus mutans mediates its cariogenicity via exopolysaccharides metabolism

2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Yalan Deng ◽  
Yingming Yang ◽  
Bin Zhang ◽  
Hong Chen ◽  
Yangyu Lu ◽  
...  
Keyword(s):  
Dental Caries ◽  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Single Species ◽  
Laser Scanning Microscopy ◽  
Gas Chromatography Mass Spectrometry ◽  
Extracellular Polysaccharides ◽  
Confocal Laser

AbstractStreptococcus mutans (S. mutans) is generally regarded as a major contributor to dental caries because of its ability to synthesize extracellular polysaccharides (EPS) that aid in the formation of plaque biofilm. The VicRKX system of S. mutans plays an important role in biofilm formation. The aim of this study was to investigate the effects of vicK gene on specific characteristics of EPS in S. mutans biofilm. We constructed single-species biofilms formed by different mutants of vicK gene. Production and distribution of EPS were detected through atomic force microscopy, scanning electron microscopy and confocal laser scanning microscopy. Microcosmic structures of EPS were analyzed by gel permeation chromatography and gas chromatography-mass spectrometry. Cariogenicity of the vicK mutant was assessed in a specific pathogen-free rat model. Transcriptional levels of cariogenicity-associated genes were confirmed by quantitative real-time polymerase chain reaction. The results showed that deletion of vicK gene suppressed biofilm formation as well as EPS production, and EPS were synthesized mostly around the cells. Molecular weight and monosaccharide components underwent evident alterations. Biofilms formed in vivo were sparse and contributed a decreased degree of caries. Moreover, expressional levels of genes related to EPS synthesis were down-regulated, except for gtfB. Our report demonstrates that vicK gene enhances biofilm formation and subsequent caries development. And this may due to its regulations on EPS metabolism, like synthesis or microcosmic features of EPS. This study suggests that vicK gene and EPS can be considered as promising targets to modulate dental caries.

scholarly journals Influence of BrpA on Critical Virulence Attributes of Streptococcus mutans

2006 ◽  
Vol 188 (8) ◽  
pp. 2983-2992 ◽  
Author(s):  
Zezhang T. Wen ◽  
Henry V. Baker ◽  
Robert A. Burne
Keyword(s):  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Tooth Surface ◽  
Confocal Laser ◽  
Carbohydrate Source ◽  
Flow Cells ◽  
Forming Characteristics

ABSTRACT Streptococcus mutans, the primary etiological agent of human dental caries, has developed multiple mechanisms to colonize and form biofilms on the tooth surface. The brpA gene codes for a predicted surface-associated protein with apparent roles in biofilm formation, autolysis, and cell division. In this study, we used two models to further characterize the biofilm-forming characteristics of a BrpA-deficient mutant, strain TW14. Compared to those of the parent strain, UA159, TW14 formed long chains and sparse microcolonies on hydroxylapatite disks but failed to accumulate and form three-dimensional biofilms when grown on glucose as the carbohydrate source. The biofilm formation defect was also readily apparent by confocal laser scanning microscopy when flow cells were used to grow biofilms. When subjected to acid killing at pH 2.8 for 45 min, the survival rate of strain TW14 was more than 1 log lower than that of the wild-type strain. TW14 was at least 3 logs more susceptible to killing by 0.2% hydrogen peroxide than was UA159. The expression of more than 200 genes was found by microarray analysis to be altered in cells lacking BrpA (P < 0.01). These results suggest that the loss of BrpA can dramatically influence the transcriptome and significantly affects the regulation of acid and oxidative stress tolerance and biofilm formation in S. mutans, which are key virulence attributes of the organism.

Candida albicans Increases Dentine Demineralization Provoked by Streptococcus mutans Biofilm

2018 ◽  
Vol 53 (3) ◽  
pp. 322-331 ◽  
Author(s):  
Aline A. Sampaio ◽  
Samilly E. Souza ◽  
Antônio P. Ricomini-Filho ◽  
Altair A. Del Bel Cury ◽  
Yuri W. Cavalcanti ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Streptococcus Mutans ◽  
Laser Scanning ◽  
Surface Hardness ◽  
Laser Scanning Microscopy ◽  
Broth Culture ◽  
Extracellular Polysaccharides ◽  
Confocal Laser ◽  
Ph Indicator ◽  
Transmission Electron

Streptococcus mutans are considered the most cariogenic bacteria, but it has been suggested that Candida albicans could increase their cariogenicity. However, the effect of this dual-species microorganisms’ combination on dentine caries has not been experimentally evaluated. Biofilms of C. albicans, S. mutans and C. albicans + S. mutans (n = 12/biofilm) were grown in ultra-filtered tryptone yeast extract broth culture medium for 96 h on root dentine slabs of known surface hardness and exposed 8 times per day for 3 min to 10% sucrose. The medium was changed 2 times per day (after the 8 cariogenic challenges and after the overnight period of famine), and aliquots were analyzed to determinate the pH (indicator of biofilm acidogenicity). After 96 h, the biofilms were collected to determine the wet weight, colony-forming units, and the amounts of extracellular polysaccharides (soluble and insoluble). Dentine demineralization was assessed by surface hardness loss (% SHL). The architecture of the biofilms was analyzed by confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM). Data were analyzed by ANOVA followed by Tukey’s test (α = 0.05). The dual-species C. albicans + S. mutans biofilm provoked higher % SHL on dentine (p < 0.05) than the S. mutans and C. albicans biofilm. This was supported by the results of biofilm acidogenicity and the amounts of soluble (6.4 ± 2.14 vs. 4.0 ± 0.94 and 1.9 ± 0.97, respectively) and insoluble extracellular polysaccharides (24.9 ± 9.22 vs. 18.9 ± 5.92 and 0.7 ± 0.48, respectively) (p < 0.05). The C. albicans biofilm alone presented low cariogenicity. The images by CLSM and TEM, respectively, suggest that the C. albicans + S. mutans biofilm is more voluminous than the S. mutans biofilm, and S. mutans cells interact with C. albicans throughout polysaccharides from the biofilm matrix. These findings show that C. albicans enhances the cariogenic potential of the S. mutans biofilm, increasing dentine demineralization.

scholarly journals Comparison of Cariogenic Characteristics between Fluoride-sensitive and Fluoride-resistant Streptococcus mutans

2020 ◽  
Vol 47 (4) ◽  
pp. 397-405
Author(s):  
Seung-Hwan Ong ◽  
Jongsoo Kim ◽  
Dong-Heon Baek ◽  
Seunghoon Yoo
Keyword(s):  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Single Species ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Rt Pcr ◽  
Oral Biofilm ◽  
Low Levels ◽  
Number Of Bacteria

The aim of this study is to compare cariogenic characteristics of fluoride-sensitive <i>Streptococcus mutans</i> [fluoride-sensitive (FS) <i>S. mutans</i> ] and fluoride-resistant <i>Streptococcus mutans</i> [fluoride-resistant (FR) <i>S. mutans</i>] in the presence of sucrose, and to evaluate its effect on cariogenic biofilm formation. <i>S. mutans</i> ATCC 25175 was continuously cultured in trypticase soy broth (TSB) containing NaF (70 ppm) for 40 days to generate FR <i>S. mutans</i> . FS and FR <i>S. mutans</i> were inoculated in TSB with or without 2% sucrose, and optical density and pH were measured every hour. An oral biofilm was formed using saliva bacteria and analyzed through confocal laser scanning microscopy and CFU count. Finally, the expression of glucosyltransferases genes of both <i>S. mutans</i> was investigated through RT-PCR. FR <i>S. mutans</i> exhibited slower growth and lower acidogenicity in the presence of sucrose compared to FS <i>S. mutans</i> . Both cariogenic and single species biofilm formation was lower in the presence of FR <i>S. mutans</i> , along with reduced number of bacteria. FR <i>S. mutans</i> showed significantly low levels of gtfB, gtfC, and gtfD expression compared to FS <i>S. mutans</i> . On the basis of results, FR <i>S. mutans</i> may be less virulent in the induction of dental caries.

scholarly journals Genetic and Physiological Effects of Noncoherent Visible Light Combined with Hydrogen Peroxide on Streptococcus mutans in Biofilm

2008 ◽  
Vol 52 (7) ◽  
pp. 2626-2631 ◽  
Author(s):  
Doron Steinberg ◽  
Daniel Moreinos ◽  
John Featherstone ◽  
Moshe Shemesh ◽  
Osnat Feuerstein
Keyword(s):  
Gene Expression ◽  
Hydrogen Peroxide ◽  
Visible Light ◽  
Streptococcus Mutans ◽  
Laser Scanning ◽  
Antibacterial Effect ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Planktonic Bacteria ◽  
Deep Layers

ABSTRACT Oral biofilms are associated with the most common infections of the oral cavity. Bacteria embedded in the biofilms are less sensitive to antibacterial agents than planktonic bacteria are. Recently, an antibacterial synergic effect of noncoherent blue light and hydrogen peroxide (H2O2) on planktonic Streptococcus mutans was demonstrated. In this study, we tested the effect of a combination of light and H2O2 on the vitality and gene expression of S. mutans embedded in biofilm. Biofilms of S. mutans were exposed to visible light (wavelengths, 400 to 500 nm) for 30 or 60 s (equivalent to 34 or 68 J/cm2) in the presence of 3 to 300 mM H2O2. The antibacterial effect was assessed by microbial counts of each treated sample compared with that of the control. The effect of light combined with H2O2 on the different layers of the biofilm was evaluated by confocal laser scanning microscopy. Gene expression was determined by real-time reverse transcription-PCR. Our results show that noncoherent light, in combination with H2O2, has a synergistic antibacterial effect through all of the layers of the biofilm. Furthermore, this treatment was more effective against bacteria in biofilm than against planktonic bacteria. The combined light and H2O2 treatment up-regulated the expression of several genes such as gtfB, brp, smu630, and comDE but did not affect relA and ftf. The ability of noncoherent visible light in combination with H2O2 to affect bacteria in deep layers of the biofilm suggests that this treatment may be applied in biofilm-related diseases as a minimally invasive antibacterial procedure.

scholarly journals Fluorescence lifetime imaging of red yeast Cystofilobasidium capitatum during growth

2018 ◽  
Vol 2 (2) ◽  
pp. 114-120 ◽  
Author(s):  
Martin Vanek ◽  
Filip Mravec ◽  
Martin Szotkowski ◽  
Dana Byrtusova ◽  
Andrea Haronikova ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Laser Scanning ◽  
Light Exposure ◽  
Laser Scanning Microscopy ◽  
Model Systems ◽  
Confocal Laser ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Red Yeast

AbstractRed yeast Cystofilobasidium capitatum autofluorescence was studied by means of confocal laser scanning microscopy (CLSM) to reveal distribution of carotenoids inside the cells. Yeasts were cultivated in 2L fermentor on glucose medium at permanent light exposure and aeration. Samples were collected at different times for CLSM, gravimetric determination of biomass and HPLC determination of pigments. To compare FLIM (Fluorescence Lifetime Imaging Microscopy) images and coupled data (obtained by CLSM) with model systems, FLIM analysis was performed on micelles of SDS:ergosterol and SDS:coenzyme Q with different content of ergosterol and coenzyme Q, respectively, and with constant addition of beta-carotene. Liposomes lecithin:ergosterol:beta-carotene were investigated too. Two different intracellular forms of carotenoids were observed during most of cultivations, with third form appeared at the beginning of stationary phase. Observed behavior is probably due to formation of some kind of carotenoid protective system in membranes of different compartments of yeast cell, especially cytoplasmic membrane.

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