Effects of brief sodium fluoride treatment on the growth of early and mature cariogenic biofilms

Antibiofilm Activity ◽

Fluoride Treatment ◽

Formation Stage ◽

Early Formation ◽

Cariogenic Biofilm

Abstract Although fluoride has been widely used in the prevention of dental caries, the effect of fluoride on the activity of biofilm in different stages of cariogenic biofilm formation is less studied. This study aimed to investigate the antibiofilm activity of sodium fluoride during early and mature Streptococcus mutans (S. mutans) biofilms 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 biofilm was treated with different concentrations of fluoride (250, 500, 1000, 2000 ppm; 5 times in total, 1 min/treatment). Acidogenicity, dry weight, colony-forming units, water-soluble/insoluble extracellular polysaccharides (EPS), and intracellular polysaccharides were analyzed and confocal laser scanning microscopy images were obtained of the two stages of biofilms (early and mature biofilms). To determine the antibiofilm activity of sodium fluoride during the formation of early and mature biofilms, and to evaluate the relationship between different concentrations of sodium fluoride and antibiofilm activity. In the early cariogenic biofilm formation stage, all fluoride concentration test groups (250, 500, 1000, 2000 ppm) significantly inhibited the growth of S. mutans biofilm. The antibiofilm and anti-EPS formation activities of the brief fluoride treatment increased in 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 the control and other fluoride treatment groups. The antimicrobial activity of fluoride is related to the formation stage of cariogenic biofilm. The early formation stage of cariogenic biofilm is more susceptible to the inhibition of fluorine than the mature stage. The fluoride treatment in the early formation stage of cariogenic biofilm may be an effective means to control the development of cariogenic biofilm and prevent caries.

Effects of brief sodium fluoride treatments on the growth of early and mature cariogenic biofilms

Scientific Reports ◽

10.1038/s41598-021-97905-0 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Ye Han

Keyword(s):

Dental Caries ◽

Sodium Fluoride ◽

Biofilm Formation ◽

Water Soluble ◽

Antibiofilm Activity ◽

Fluoride Treatment ◽

Biofilm Development ◽

Cariogenic Biofilm

AbstractAlthough 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.

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

PLoS ONE ◽

10.1371/journal.pone.0259895 ◽

2021 ◽

Vol 16 (11) ◽

pp. e0259895

Author(s):

Ye Han

Keyword(s):

Cigarette Smoking ◽

Streptococcus Mutans ◽

Water Soluble ◽

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.

Quaternized Chitosan InhibitsicaATranscription and Biofilm Formation byStaphylococcuson a Titanium Surface

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01005-10 ◽

2010 ◽

Vol 55 (2) ◽

pp. 860-866 ◽

Cited By ~ 68

Author(s):

Zhao-Xiang Peng ◽

Bing Tu ◽

Yang Shen ◽

Lin Du ◽

Ling Wang ◽

...

Keyword(s):

Staphylococcus Epidermidis ◽

Biofilm Formation ◽

Laser Scanning ◽

Titanium Surface ◽

Water Soluble ◽

Confocal Laser ◽

Trimethylammonium Chloride ◽

Orthopedic Implant

ABSTRACTOur previous study (Z. X. Peng et al., Carbohydr. Polym.81:275-283, 2010) demonstrated that water-soluble quaternary ammonium salts, which are produced by the reaction of chitosan with glycidyl trimethylammonium chloride, provide chitosan derivatives with enhanced antibacterial ability. Because biofilm formation is believed to comprise the key step in the development of orthopedic implant-related infections, we further evaluated the efficacy of hydroxypropyltrimethyl ammonium chloride chitosan (HACC) with different degrees of substitution (DS; referred to as HACC 6%, 18%, and 44%) in preventing biofilm formation on a titanium surface. We used a tissue culture plate method to quantify the biomass ofStaphylococcus epidermidisandStaphylococcus aureusbiofilms and found that HACC, especially HACC 18% and 44%, significantly inhibited biofilm formation compared to the untreated control, even at concentrations far below their MICs (P< 0.05). Scanning electron microscopy showed that inhibition of biofilm formation on titanium increased dramatically with increased DS and HACC concentrations. Confocal laser scanning microscopy indicated that growth of a preexisting biofilm on titanium was inhibited by concentrations of HACC 18% and 44% below their minimum biofilm eradication concentrations. We also demonstrated that HACC inhibited the expression oficaA, which mediates the production of extracellular polysaccharides, both in new biofilms and in preexisting biofilms on titanium. Our results indicate that HACC may serve as a new antibacterial agent to inhibit biofilm formation and prevent orthopedic implant-related infections.

Confocal and SEM imaging to demonstrate food pathogen- biofilm biocontrol by pyocyanin from Pseudomonas aeruginosa BTRY1

International Journal of Bioassays ◽

10.21746/ijbio.2017.01.007 ◽

2016 ◽

Vol 6 (01) ◽

pp. 5218

Author(s):

Laxmi Mohandas ◽

Anju T. R. ◽

Sarita G. Bhat*

Keyword(s):

Pseudomonas Aeruginosa ◽

Biofilm Formation ◽

Laser Scanning ◽

Confocal Laser ◽

Antibiofilm Activity ◽

Opportunistic Pathogens ◽

Redox Active ◽

Sem Imaging ◽

The Impact

An assortment of redox-active phenazine compounds like pyocyanin with their characteristic blue-green colour are synthesized by Pseudomonas aeruginosa, Gram-negative opportunistic pathogens, which are also considered one of the most commercially valuable microorganisms. In this study, pyocyanin from Pseudomonas aeruginosa BTRY1 from food sample was assessed for its antibiofilm activity by micro titer plate assay against strong biofilm producers belonging to the genera Bacillus, Staphylococcus, Brevibacterium and Micrococcus. Pyocyanin inhibited biofilm activity in very minute concentrations. This was also confirmed by Scanning Electron Microscopy (SEM) and Confocal Laser Scanning Microscopy (CLSM). Both SEM and CLSM helped to visualize the biocontrol of biofilm formation by eight pathogens. The imaging and quantification by CLSM also established the impact of pyocyanin on biofilm-biocontrol mainly in the food industry.

Antibiofilm Properties of Temporin-L on Pseudomonas fluorescens in Static and In-Flow Conditions

International Journal of Molecular Sciences ◽

10.3390/ijms21228526 ◽

2020 ◽

Vol 21 (22) ◽

pp. 8526

Author(s):

Angela Di Somma ◽

Federica Recupido ◽

Arianna Cirillo ◽

Alessia Romano ◽

Alessandra Romanelli ◽

...

Keyword(s):

Pseudomonas Fluorescens ◽

Biofilm Formation ◽

Confocal Laser ◽

Antibiofilm Activity ◽

Dead Cell ◽

Flow Conditions ◽

Dynamic Conditions ◽

Ability To Act ◽

Static Conditions

Biofilms consist of a complex microbial community adhering to biotic or abiotic surfaces and enclosed within a protein/polysaccharide self-produced matrix. The formation of this structure represents the most important adaptive mechanism that leads to antibacterial resistance, and therefore, closely connected to pathogenicity. Antimicrobial peptides (AMPs) could represent attractive candidates for the design of new antibiotics because of their specific characteristics. AMPs show a broad activity spectrum, a relative selectivity towards their targets (microbial membranes), the ability to act on both proliferative and quiescent cells, a rapid mechanism of action, and above all, a low propensity for developing resistance. This article investigates the effect at subMIC concentrations of Temporin-L (TL) on biofilm formation in Pseudomonas fluorescens (P. fluorescens) both in static and dynamic conditions, showing that TL displays antibiofilm properties. Biofilm formation in static conditions was analyzed by the Crystal Violet assay. Investigation of biofilms in dynamic conditions was performed in a commercial microfluidic device consisting of a microflow chamber to simulate real flow conditions in the human body. Biofilm morphology was examined using Confocal Laser Scanning Microscopy and quantified via image analysis. The investigation of TL effects on P. fluorescens showed that when subMIC concentrations of this peptide were added during bacterial growth, TL exerted antibiofilm activity, impairing biofilm formation both in static and dynamic conditions. Moreover, TL also affects mature biofilm as confocal microscopy analyses showed that a large portion of preformed biofilm architecture was clearly perturbed by the peptide addition with a significative decrease of all the biofilm surface properties and the overall biomass. Finally, in these conditions, TL did not affect bacterial cells as the live/dead cell ratio remained unchanged without any increase in damaged cells, confirming an actual antibiofilm activity of the peptide.

The Antibiofilm Effect of a Medical Device Containing TIAB on Microorganisms Associated with Surgical Site Infection

Molecules ◽

10.3390/molecules24122280 ◽

2019 ◽

Vol 24 (12) ◽

pp. 2280 ◽

Cited By ~ 2

Author(s):

Valentina Puca ◽

Tonino Traini ◽

Simone Guarnieri ◽

Simone Carradori ◽

Francesca Sisto ◽

...

Keyword(s):

Medical Device ◽

Biofilm Formation ◽

Inhibitory Concentration ◽

Test Method ◽

Antibiofilm Activity ◽

E Coli ◽

Post Surgery ◽

Agar Diffusion Test ◽

Surgical Sutures

Surgical site infections (SSIs) represent the most common nosocomial infections, and surgical sutures are optimal surfaces for bacterial adhesion and biofilm formation. Staphylococcus spp., Enterococcus spp., and Escherichia coli are the most commonly isolated microorganisms. The aim of this research was to evaluate the antibiofilm activity of a medical device (MD) containing TIAB, which is a silver-nanotech patented product. The antibacterial effect was evaluated against Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212, and E. coli ATCC 25922 by assessing the minimum inhibitory concentration (MIC) by the Alamar Blue® (AB) assay. The antibiofilm effect was determined by evaluation of the minimum biofilm inhibitory concentration (MBIC) and colony-forming unit (CFU) count. Subsequently, the MD was applied on sutures exposed to the bacterial species. The antimicrobial and antibiofilm effects were evaluated by the agar diffusion test method, confocal laser scanning microscopy (CLSM), and scanning electron microscopy (SEM). The MIC was determined for S. aureus and E. faecalis at 2 mg/mL, while the MBIC was 1.5 mg/mL for S. aureus and 1 mg/mL for E. faecalis. The formation of an inhibition zone around three different treated sutures confirmed the antimicrobial activity, while the SEM and CLSM analysis performed on the MD-treated sutures underlined the presence of a few adhesive cells, which were for the most part dead. The MD showed antimicrobial and antibiofilm activities versus S. aureus and E. faecalis, but a lower efficacy against E. coli. Surgical sutures coated with the MD have the potential to reduce SSIs as well as the risk of biofilm formation post-surgery.

Antibiofilm activity of antifungal drugs, including the novel drug olorofim, against Lomentospora prolificans

Journal of Antimicrobial Chemotherapy ◽

10.1093/jac/dkaa157 ◽

2020 ◽

Cited By ~ 1

Author(s):

Lisa Kirchhoff ◽

Silke Dittmer ◽

Ann-Kathrin Weisner ◽

Jan Buer ◽

Peter-Michael Rath ◽

...

Keyword(s):

Amphotericin B ◽

Biofilm Formation ◽

Laser Scanning ◽

Antifungal Agents ◽

Fungal Infections ◽

Pathogenic Fungus ◽

Antifungal Drugs ◽

Confocal Laser ◽

Antibiofilm Activity

Abstract Objectives Patients with immunodeficiency or cystic fibrosis frequently suffer from respiratory fungal infections. In particular, biofilm-associated fungi cause refractory infection manifestations, linked to increased resistance to anti-infective agents. One emerging filamentous fungus is Lomentospora prolificans. Here, the biofilm-formation capabilities of L. prolificans isolates were investigated and the susceptibility of biofilms to various antifungal agents was analysed. Methods Biofilm formation of L. prolificans (n = 11) was estimated by crystal violet stain and antibiofilm activity was additionally determined via detection of metabolically active biofilm using an XTT assay. Amphotericin B, micafungin, voriconazole and olorofim were compared with regard to their antibiofilm effects when added prior to adhesion, after adhesion and on mature and preformed fungal biofilms. Imaging via confocal laser scanning microscopy was carried out to demonstrate the effect of drug treatment on the fungal biofilm. Results Antibiofilm activities of the tested antifungal agents were shown to be most effective on adherent cells whilst mature biofilm was the most resistant. The most promising antibiofilm effects were detected with voriconazole and olorofim. Olorofim showed an average minimum biofilm eradication concentration (MBEC) of 0.06 mg/L, when added prior to and after adhesion. The MBECs of voriconazole were ≤4 mg/L. On mature biofilm the MBECs of olorofim and voriconazole were higher than the previously determined MICs against planktonic cultures. In contrast, amphotericin B and especially micafungin did not exhibit sufficient antibiofilm activity against L. prolificans. Conclusions To our knowledge, this is the first study demonstrating the antibiofilm potential of olorofim against the human pathogenic fungus L. prolificans.

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

10.21203/rs.3.rs-221658/v1 ◽

2021 ◽

Author(s):

Ye Han

Keyword(s):

Treatment Group ◽

Cigarette Smoking ◽

Streptococcus Mutans ◽

Laser Scanning ◽

Water Soluble ◽

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.

Effect of LongZhang Gargle on Biofilm Formation and Acidogenicity ofStreptococcus mutans In Vitro

BioMed Research International ◽

10.1155/2016/5829823 ◽

2016 ◽

Vol 2016 ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

Yutao Yang ◽

Shiyu Liu ◽

Yuanli He ◽

Zhu Chen ◽

Mingyun Li

Keyword(s):

Dental Caries ◽

Biofilm Formation ◽

Acid Production ◽

Hot Spot ◽

Bacterial Species ◽

Chinese Herbs ◽

Minimum Concentration ◽

Planktonic Growth

Streptococcus mutans, with the ability of high-rate acid production and strong biofilm formation, is considered the predominant bacterial species in the pathogenesis of human dental caries. Natural products which may be bioactive againstS. mutanshave become a hot spot to researches to control dental caries. LongZhang Gargle, completely made from Chinese herbs, was investigated for its effects on acid production and biofilm formation byS. mutansin this study. The results showed an antimicrobial activity of LongZhang Gargle againstS. mutansplanktonic growth at the minimum inhibitory concentration (MIC) of 16% and minimum bactericidal concentration (MBC) of 32%. Acid production was significantly inhibited at sub-MIC concentrations. Biofilm formation was also significantly disrupted, and 8% was the minimum concentration that resulted in at least 50% inhibition of biofilm formation (MBIC50). A scanning electron microscopy (SEM) showed an effective disruption of LongZhang Gargle onS. mutansbiofilm integrity. In addition, a confocal laser scanning microscopy (CLSM) suggested that the extracellular polysaccharides (EPS) synthesis could be inhibited by LongZhang Gargle at a relatively low concentration. These findings suggest that LongZhang Gargle may be a promising natural anticariogenic agent in that it suppresses planktonic growth, acid production, and biofilm formation againstS. mutans.

A Surface-Active Agent from Saccharomyces cerevisiae Influences Staphylococcal Adhesion and Biofilm Development

Zeitschrift für Naturforschung C ◽

10.1515/znc-2007-5-618 ◽

2007 ◽

Vol 62 (5-6) ◽

pp. 433-438 ◽

Cited By ~ 7

Author(s):

Elzbieta Walencka ◽

Marzena Wieckowska-Szakiel ◽

Sylwia Rozalska ◽

Beata Sadowska ◽

Barbara Rozalska

Keyword(s):

Saccharomyces Cerevisiae ◽

Biofilm Formation ◽

Active Agent ◽

Cell Surface Hydrophobicity ◽

Water Soluble ◽

Confocal Laser ◽

Tetrazolium Salt ◽

Initial Adhesion ◽

Biofilm Development

Bacterial biofilms which are responsible for a number of diseases are very difficult to control effectively because of their high resistance to antibiotics and the host defence system. The use of natural products decreasing or preventing initial adhesion of bacteria and biofilm formation is one of the alternative therapeutic strategies taken into consideration. We ask the question, whether a crude extract from the cell wall of Saccharomyces cerevisiae (mannoprotein), which possesses surfactant activity, may be used as inhibitor of Staphylococcus aureus and S. epidermidis biofilm development. By using the “bactericidal spot assay” it was demonstrated that mannoprotein had no direct antibiotic activity against the tested strains. The influence of this extract on initial adhesion, biofilm formation and dispersal of preformed biofilms was studied using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. In this assay, live bacteria with an active electron transport system reduce the tetrazolium salt to a water-soluble purple formazan product, and optical density reading (A550) values are directly dependent on their cell numbers. Yeast-derived surfactant, when adsorbed in the microplate wells or present in the medium, was effective both in decreasing the initial deposition of staphylococci and in reducing the amount of growing biofilm, quantitated after 24 h of co-incubation with the bacteria. It also changed the parameters of biofilm morphology analyzed by PHLIP - the confocal laser scanning microscopy image quantification package. Mannoprotein also accelerated the detachment of mature staphylococcal biofilms, preformed in optimal conditions. It was concluded that mannoprotein anti-biofilm action reflects its influence on cell surface hydrophobicity.