scholarly journals Effect of LongZhang Gargle on Dual-Species Biofilm of Candida albicans and Streptococcus mutans

2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Jinglei Gong ◽  
Die Hu ◽  
Jinzhi He ◽  
Ling Zou ◽  
Zhu Chen ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Streptococcus Mutans ◽  
Hot Spot ◽  
Fungal Spores ◽  
Cell Activity ◽  
Single Species ◽  
Laser Scanning Microscopy ◽  
Oral Disease ◽  
Extracellular Polysaccharides ◽  
Confocal Laser

Bioactive natural products have become a hot spot for oral disease treatments. At the present study, LongZhang Gargle was investigated for its effects on single-species biofilms of Candida albicans and dual-species biofilms of Candida albicans and Streptococcus mutans. Two different models of single and dual-species biofilms were grown in YNBB medium under appropriate conditions. Biofilm biomass, biofilm architecture, and cell activity in biofilms were assessed using Crystal Violet Staining, MTT, scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM). Significant reductions of biofilm biomass and fungus activity were obtained when treated with LongZhang Gargle at 2% ( P < 0.05 ), 4% ( P < 0.05 ), and 8% ( P < 0.05 ) in single-species biofilms of C. albicans, and at 4% ( P < 0.05 ) and 8% ( P < 0.05 ) in double-species biofilms. Suppression of density, thickness, and the proportion of hyphae and fungal spores were obtained under SEM and CLSM. In conclusion, LongZhang Gargle affects single and dual-species biofilms by inhibiting biofilm biomass, cell activity, and formation of hyphae, but it does not affect the production of Extracellular polysaccharides (EPS). We speculate that LongZhang Gargle would be a promising natural drug, which can be used in treatment against C. albicans and S. mutans in oral diseases.

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.

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 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 Effects of cigarette smoking on the growth of Streptococcus mutans biofilms: An in vitro study

PLoS ONE ◽  
2021 ◽  
Vol 16 (11) ◽  
pp. e0259895
Author(s):  
Ye Han
Keyword(s):  
Cigarette Smoking ◽  
Streptococcus Mutans ◽  
Water Soluble ◽  
Laser Scanning Microscopy ◽  
Extracellular Polysaccharides ◽  
Confocal Laser ◽  
Control Group ◽  
Exposure Group ◽  
Cariogenic Biofilm

The increased incidence of dental caries by cigarette smoking (CS) has been widely reported in epidemiological studies, but the relationship between CS and cariogenic biofilm growth has been rarely studied. This study aims to investigate the effects of CS exposure on the growth and virulence of Streptococcus mutans biofilms (S. mutans). Briefly, S. mutans biofilms were formed on saliva-coated hydroxyapatite disks, which were exposed to CS 1, 3, and 6 times per day, respectively. In addition, S. mutans biofilms without CS exposure were considered as the control group. Acidogenicity, dry weight, colony-forming units (CFUs), water-soluble/insoluble extracellular polysaccharides (EPSs), and intracellular polysaccharides (IPSs) were analyzed and confocal laser scanning microscopy (CLSM) images of 74-h-old S. mutans biofilms were obtained. The lowest accumulation of biofilms and EPSs were detected in the 6 times/day CS exposure group compared with those of the control group and other CS exposure groups in 74-h-old S. mutans biofilms. CLSM also revealed the lowest bacterial count (live and dead cells) and EPSs biovolume in the six times/day CS exposure group in 74-h-old S. mutans biofilms. CS exposure inhibited the growth of S. mutans biofilm in vitro study, the anti-cariogenic biofilm formation was enhanced with a dose (frequency)-dependent at which frequency has more influence in the present findings.

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 Low-Temperature Plasma as an Approach for Inhibiting a Multi-Species Cariogenic Biofilm

2021 ◽  
Vol 11 (2) ◽  
pp. 570
Author(s):  
Leandro W. Figueira ◽  
Beatriz H. D. Panariello ◽  
Cristiane Y. Koga-Ito ◽  
Simone Duarte
Keyword(s):  
Low Temperature ◽  
Laser Scanning ◽  
Single Species ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Low Temperature Plasma ◽  
Temperature Plasma ◽  
Plasma Device ◽  
Negative Controls ◽  
Promising Source

This study aimed to determine how low-temperature plasma (LTP) treatment affects single- and multi-species biofilms formed by Streptococcus mutans, Streptococcus sanguinis, and Streptococcus gordonii formed on hydroxyapatite discs. LTP was produced by argon gas using the kINPen09™ (Leibniz Institute for Plasma Science and Technology, INP, Greifswald, Germany). Biofilms were treated at a 10 mm distance from the nozzle of the plasma device to the surface of the biofilm per 30 s, 60 s, and 120 s. A 0.89% saline solution and a 0.12% chlorhexidine solution were used as negative and positive controls, respectively. Argon flow at three exposure times (30 s, 60 s, and 120 s) was also used as control. Biofilm viability was analyzed by colony-forming units (CFU) recovery and confocal laser scanning microscopy. Multispecies biofilms presented a reduction in viability (log10 CFU/mL) for all plasma-treated samples when compared to both positive and negative controls (p < 0.0001). In single-species biofilms formed by either S. mutans or S. sanguinis, a significant reduction in all exposure times was observed when compared to both positive and negative controls (p < 0.0001). For single-species biofilms formed by S. gordonii, the results indicate total elimination of S. gordonii for all exposure times. Low exposure times of LTP affects single- and multi-species cariogenic biofilms, which indicates that the treatment is a promising source for the development of new protocols for the control of dental caries.

scholarly journals Effect of antibacterial photodynamic therapy on Streptococcus mutans plaque biofilm in vitro

2020 ◽  
Vol 13 (06) ◽  
pp. 2050022
Author(s):  
Xiaoyue Liang ◽  
Zhaohui Zou ◽  
Zheng Zou ◽  
Changyi Li ◽  
Xiaoxi Dong ◽  
...  
Keyword(s):  
Photodynamic Therapy ◽  
Lactic Acid ◽  
Streptococcus Mutans ◽  
Antibacterial Effect ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Saline Control ◽  
Control Group ◽  
Plaque Biofilm

The main objective of this study is to evaluate the antibacterial effect of antibacterial photodynamic therapy (aPDT) on Streptococcus mutans (S. mutans) biofilm model in vitro. The selection of photosensitizers is the key step for the efficacy of photodynamic therapy (PDT). However, no studies have been conducted in the oral field to compare the functional characteristics and application effects of PDT mediated by various photosensitizers. In this research, the antibacterial effect of Methylene blue (MB)/650[Formula: see text]nm laser and Hematoporphyrin monomethyl ether (HMME)/532[Formula: see text]nm laser on S. mutans biofilm was compared under different energy densities to provide experimental reference for the clinical application of the two PDT. The yield of lactic acid was analyzed by Colony forming unit (CFU) and spectrophotometry, and the complete biofilm activity was measured by Confocal Laser Scanning Microscopy (CLSM) to evaluate the bactericidal effect on each group. Based on the results of CFU, the bacterial colonies formed by 30.4[Formula: see text]J/cm2 532[Formula: see text]nm MB-aPDT group and 30.4[Formula: see text]J/cm2 532[Formula: see text]nm HMME-aPDT group were significantly less than those in other groups, and the bacterial colonies in HMME-aPDT group were less than those in HMME-aPDT group. Lactic acid production in all treatment groups except the photosensitizer group was statistically lower than that in the normal saline control group. The activity of bacterial plaque biofilm was significantly decreased in the two groups treated with 30.4[Formula: see text]J/cm2 aPDT. Therefore, aPDT suitable for energy measurement can kill S. mutans plaque biofilm, and MB-aPDT is better than HMME-aPDT.

scholarly journals Symbiotic Relationship between Streptococcus mutans and Candida albicans Synergizes Virulence of Plaque BiofilmsIn Vivo

2014 ◽  
Vol 82 (5) ◽  
pp. 1968-1981 ◽  
Author(s):  
Megan L. Falsetta ◽  
Marlise I. Klein ◽  
Punsiri M. Colonne ◽  
Kathleen Scott-Anne ◽  
Stacy Gregoire ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Dental Caries ◽  
Streptococcus Mutans ◽  
Bacterial Pathogen ◽  
Single Species ◽  
Content Type ◽  
3 Dimensional ◽  
Biofilm Development

ABSTRACTStreptococcus mutansis often cited as the main bacterial pathogen in dental caries, particularly in early-childhood caries (ECC).S. mutansmay not act alone;Candida albicanscells are frequently detected along with heavy infection byS. mutansin plaque biofilms from ECC-affected children. It remains to be elucidated whether this association is involved in the enhancement of biofilm virulence. We showed that the ability of these organisms together to form biofilms is enhancedin vitroandin vivo. The presence ofC. albicansaugments the production of exopolysaccharides (EPS), such that cospecies biofilms accrue more biomass and harbor more viableS. mutanscells than single-species biofilms. The resulting 3-dimensional biofilm architecture displays sizeableS. mutansmicrocolonies surrounded by fungal cells, which are enmeshed in a dense EPS-rich matrix. Using a rodent model, we explored the implications of this cross-kingdom interaction for the pathogenesis of dental caries. Coinfected animals displayed higher levels of infection and microbial carriage within plaque biofilms than animals infected with either species alone. Furthermore, coinfection synergistically enhanced biofilm virulence, leading to aggressive onset of the disease with rampant carious lesions. Ourin vitrodata also revealed that glucosyltransferase-derived EPS is a key mediator of cospecies biofilm development and that coexistence withC. albicansinduces the expression of virulence genes inS. mutans(e.g.,gtfB,fabM). We also found thatCandida-derived β1,3-glucans contribute to the EPS matrix structure, while fungal mannan and β-glucan provide sites for GtfB binding and activity. Altogether, we demonstrate a novel mutualistic bacterium-fungus relationship that occurs at a clinically relevant site to amplify the severity of a ubiquitous infectious disease.

scholarly journals Antibody response to the 45 kDa Candida albicans antigen in an animal model and potential role of the antigen in adherence

2008 ◽  
Vol 57 (12) ◽  
pp. 1466-1472 ◽  
Author(s):  
Helena Bujdáková ◽  
Ema Paulovičová ◽  
Silvia Borecká-Melkusová ◽  
Juraj Gašperík ◽  
Soňa Kucharíková ◽  
...  
Keyword(s):  
Animal Model ◽  
Candida Albicans ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Vaccine Development ◽  
Laser Scanning Microscopy ◽  
Model Systems ◽  
Confocal Laser

The Candida antigen CR3-RP (complement receptor 3-related protein) is supposed to be a ‘mimicry’ protein because of its ability to bind antibody directed against the α subunit of the mammalian CR3 (CD11b/CD18). This study aimed to (i) investigate the specific humoral isotypic response to immunization with CR3-RP in vivo in a rabbit animal model, and (ii) determine the role of CR3-RP in the adherence of Candida albicans in vitro using the model systems of buccal epithelial cells (BECs) and biofilm formation. The synthetic C. albicans peptide DINGGGATLPQ corresponding to 11 amino-acids of the CR3-RP sequence DINGGGATLPQALXQITGVIT, determined by N-terminal sequencing, was used for immunization of rabbits to obtain polyclonal anti-CR3-PR serum and for subsequent characterization of the humoral isotypic response of rabbits. A significant increase of IgG, IgA and IgM anti-CR3-RP specific antibodies was observed after the third (P<0.01) and the fourth (P<0.001) immunization doses. The elevation of IgA levels suggested peptide immunomodulation of the IgA1 subclass, presumably in coincidence with Candida epithelial adherence. Blocking CR3-RP with polyclonal anti-CR3-RP serum reduced the ability of Candida to adhere to BECs, in comparison with the control, by up to 35 % (P<0.001), and reduced biofilm formation by 28 % (P<0.001), including changes in biofilm thickness and integrity detected by confocal laser scanning microscopy. These properties of CR3-RP suggest that it has potential for future vaccine development.

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