Antimicrobial effect of toothbrush with light emitting diode on dental biofilm attached to zirconia surface: an in vitro study

PURPOSE: The purpose of this study was to evaluate the antimicrobial effects of a toothbrush with light-emitting diodes (LEDs) on periodontitis-associated dental biofilm attached to a zirconia surface by static and dynamic methods. MATERIALS AND METHODS: Zirconia disks (12 mm diameter, 2.5 mm thickness) were inserted into a 24-well plate (static method) or inside a Center for Disease Control and Prevention (CDC) biofilm reactor (dynamic method) to form dental biofilms using Streptococcus gordonii and Fusobacterium nucleatum. The disks with biofilm were subdivided into five treatment groups-control, commercial photodynamic therapy (PDT), toothbrush alone (B), brush with LED (BL), and brush with LED+erythrosine (BLE). After treatment, the disks were agitated to detach the bacteria, and the resulting solutions were spread directly on selective agar. The number of viable bacteria and percentage of bacterial reduction were determined from colony counts. Scanning electron microscopy (SEM) was performed to visualize alterations in bacterial morphology. RESULTS: No significant difference in biofilm formation was observed between dynamic and static methods. A significant difference was observed in the number of viable bacteria between the control and all experimental groups (P < 0.05). The percentage of bacterial reduction in the BLE group was significantly higher than in the other treated groups (P < 0.05). SEM revealed damaged bacterial cell walls in the PDT, BL, and BLE groups, but intact cell walls in the control and B groups. CONCLUSION: The findings suggest that an LED toothbrush with erythrosine is more effective than other treatments in reducing the viability of periodontitis-associated bacteria attached to zirconia in vitro.

Adhesion and Biofilm Formation Abilities of Bacteria Isolated from Dental Unit Waterlines / 치위생과학회지

The purpose of our study is to compare the adhesion and biofilm formation abilities of isolates from water discharged from dental unit waterlines (DUWLs). Bacteria were isolated from a total of 15 DUWLs. Twelve isolates were selected for the experiment. To confirm the adhesion ability of the isolates, each isolate was attached to a glass coverslip using a 12-well plate. Plates were incubated at 26℃ for 7 days, and the degree of adhesion of each isolate was scored. To verify the biofilm formation ability of each isolate, biofilms were allowed to form on a 96-well polystyrene flat-bottom microtiter plate. The biofilm accumulations of all isolates formed at 26℃ for 7 days were identified and compared. A total of 56 strains were isolated from 15 water samples including 12 genera and 31 species. Of the 56 isolates, 12 isolates were selected according to the genus and used in the experiment. Sphingomonas echinoides, Methylobacterium aquaticum, and Cupriavidus pauculus had the highest adhesion ability scores of +3 among 12 isolates. Among these three isolates, the biofilm accumulation of C. pauculus was the highest and that of S. echinoides was the third-most abundant. The lowest biofilm accumulations were identified in Microbacterium testaceum and M. aquaticum. Most isolates with high adhesion ability also exhibited high biofilm formation ability. Analysis of adhesion and biofilm formation of the isolates from DUWLs can provide useful information to understand the mechanism of DUWL biofilm formation and development.

Efficacy of an LED toothbrush on a Porphyromonas gingivalis biofilm on a sandblasted and acid-etched titanium surface: an in vitro study

PURPOSE: The aim of this study was to evaluate the antimicrobial effect of a newly devised toothbrush with light-emitting diodes (LEDs) on Porphyromonas gingivalis attached to sandblasted and acid-etched titanium surfaces. METHODS: The study included a control group, a commercial photodynamic therapy (PDT) group, and 3 test groups (B, BL, and BLE). The disks in the PDT group were placed in methylene blue and then irradiated with a diode laser. The B disks were only brushed, the BL disks were brushed with an LED toothbrush, and the BLE disks were placed into erythrosine and then brushed with an LED toothbrush. After the different treatments, bacteria were detached from the disks and spread on selective agar. The number of viable bacteria and percentage of bacterial reduction were determined from colony counts. Scanning electron microscopy was performed to visualize bacterial alterations. RESULTS: The number of viable bacteria in the BLE group was significantly lower than that in the other groups (P < 0.05). Scanning electron microscopy showed that bacterial cell walls were intact in the control and B groups, but changed after commercial PDT and LED exposure. CONCLUSIONS: The findings suggest that an LED toothbrush with erythrosine treatment was more effective than a commercial PDT kit in reducing the number of P. gingivalis cells attached to surface-modified titanium in vitro.

Establishment of a Dental Unit Biofilm Model Using Well-Plate / 치위생과학회지

The water discharged from dental unit waterlines (DUWLs) is heavily contaminated with bacteria. The development of efficient disinfectants is required to maintain good quality DUWL water. The purpose of this study was to establish a DUWL biofilm model using well-plates to confirm the effectiveness of disinfectants in the laboratory. Bacteria were obtained from the water discharged from DUWLs and incubated in R2A liquid medium for 10 days. The bacterial solution cultured for 10 days was made into stock and these stocks were incubated in R2A broth and batch mode for 5 days. Batch-cultured bacterial culture solution and polyurethane tubing sections were incubated in 12-well plates for 4 days. Biofilm accumulation was confirmed through plating on R2A solid medium. In addition, the thickness of the biofilm and the shape and distribution of the constituent bacteria were confirmed using confocal laser microscopy and scanning electron microscopy. The average accumulation of the cultured biofilm over 4 days amounted to 1.15×10⁷ CFU/cm². The biofilm was widely distributed on the inner surface of the polyurethane tubing and consisted of cocci, short-length rods and medium-length rods. The biofilm thickness ranged from 2 µm to 7 µm. The DUWL biofilm model produced in this study can be used to develop disinfectants and study DUWL biofilm-forming bacteria.

A Case of Osteoblastoma Accompanied with Concha Bullosa of the Inferior Turbinate / 대한이비인후과학회지

Osteoblastoma is a rare benign tumor, representing less than 1% of all bone tumors. Most cases occur in the vertebrae and in the long bones. Intranasal or paranasal osteoblastoma is particularly rare and only one case of osteoblastoma in the inferior turbinate has been reported in the world literature. Treatment is intralesional curettage or en bloc resection. Since the tumor is benign, conservative surgery is curative in about 80-90% of the cases. Concha bullosa is an abnormal pneumatization of the intranasal turbinates and inferior concha bullosa is a very rare condition. We report an unusual case of osteoblastoma occurring together with inferior concha bullosa. The tumor and inferior concha bullosa were removed by endoscopic submucosal inferior turbinoplasty, with favorable results. Related articles are reviewed and brief discussions are presented in regards to the case findings.

Comparison of periodontitis-associated oral biofilm formation under dynamic and static conditions

PURPOSE: The purpose of this study was to compare the characteristics of single- and dual-species in vitro oral biofilms made by static and dynamic methods. METHODS: Hydroxyapatite (HA) disks, 12.7 mm in diameter and 3 mm thick, were coated with processed saliva for 4 hours. The disks were divided into a static method group and a dynamic method group. The disks treated with a static method were cultured in 12-well plates, and the disks in the dynamic method group were cultured in a Center for Disease Control and Prevention (CDC) biofilm reactor for 72 hours. In the single- and dual-species biofilms, Fusobacterium nucleatum and Porphyromonas gingivalis were used, and the amount of adhering bacteria, proportions of species, and bacterial reduction of chlorhexidine were examined. Bacterial adhesion was examined with scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). RESULTS: Compared with the biofilms made using the static method, the biofilms made using the dynamic method had significantly lower amounts of adhering and looser bacterial accumulation in SEM and CLSM images. The proportion of P. gingivalis was higher in the dynamic method group than in the static method group; however, the difference was not statistically significant. Furthermore, the biofilm thickness and bacterial reduction by chlorhexidine showed no significant differences between the 2 methods. CONCLUSIONS: When used to reproduce periodontal biofilms composed of F. nucleatum and P. gingivalis, the dynamic method (CDC biofilm reactor) formed looser biofilms containing fewer bacteria than the well plate. However, this difference did not influence the thickness of the biofilms or the activity of chlorhexidine. Therefore, both methods are useful for mimicking periodontitis-associated oral biofilms.

In vitro antimicrobial activity of different mouthwashes available in Korea / 대한구강보건학회지

OBJECTIVES: The purpose of this study was to compare the oral antimicrobial effects of seven different mouthwashes available in Korea. METHODS: To examine the antimicrobial effects of the seven mouthwashes, their minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were determined using broth microdilution methods. Streptococcus mutans ATCC 25175, Lactobacillus acidophilus ATCC 4355, Candida albicans KCTC 7270, and Porphyromonas gingivalis ATCC 33277 were used in this experiment. S. mutans and P. gingivalis were examined using a scanning electron microscope (SEM), after treatment with the mouthwashes containing cetylpyridinium chloride (CPC). RESULTS: Mouthwashes containing CPC had lower MIC and MBC values against the four microorganisms. Their bactericidal effects were concentration-dependent. S. mutans and C. albicans were highly sensitive to the concentration of CPC in the mouthwashes. According to the SEM observation, the treatment of bacteria with mouthwashes containing CPC, changed the cell surface texture of S. mutans and P. gingivalis. CONCLUSIONS: Mouthwashes containing CPC showed relatively lower MIC and MBC values under the same conditions against the four microorganisms used in this study.

Developing a Dental Unit Waterline Model Using General Laboratory Equipments / 치위생과학회지

Water supplied through dental unit waterlines (DUWLs) has been shown to contain high number of bacteria. To reduce the contamination of DUWLs, it is essential to develop effective disinfectants. It is, however, difficulty to obtain proper DUWL samples for studies. The purpose of this study was to establish a simple laboratory model for reproducing DUWL biofilms. The bacteria obtained from DUWLs were cultured in R2A liquid medium for 10 days, and then stored at −70℃. This stock was inoculated into R2A liquid medium and incubated in batch mode. After 5 days of culturing, it was inoculated into the biofilm formation model developed in this study. Our biofilm formation model comprised of a beaker containing R2A liquid medium and five glass rods attached to DUWL polyurethane tubing. Biofilm was allowed to form on the stir plate and the medium was replaced every 2 days. After 4 days of biofilm formation in the laboratory model, biofilm thickness, morphological characteristics and distribution of the composing bacteria were examined by confocal laser microscopy and scanning electron microscopy. The mean of biofilm accumulation was 4.68×10⁴ colony forming unit/cm² and its thickness was 10~14 µm. In our laboratory model, thick bacterial lumps were observed in some parts of the tubing. To test the suitability of this biofilm model system, the effectiveness of disinfectants such as sodium hypochlorite, hydrogen peroxide, and chlorhexidine, was examined by their application to the biofilm formed in our model. Lower concentrations of disinfectants were less effective in reducing the count of bacteria constituting the biofilm. These results showed that our DUWL biofilm laboratory model was appropriate for comparison of disinfectant effects. Our laboratory model is expected to be useful for various other purposes in further studies.

Effect of Sub-minimal Inhibitory Concentration of Chlorhexidine on Biofilm Formation and Coaggregation of Early Colonizers, Streptococci and Actinomycetes

Chlorhexidine has long been used in mouth washes for the control of dental caries, gingivitis and dental plaque. Minimal inhibitory concentration (MIC) is the lowest concentration of an antimicrobial substance to inhibit the growth of bacteria. Concentrations lower than the MIC are called sub minimal inhibitory concentrations (sub-MICs). Many studies have reported that sub-MICs of antimicrobial substances can affect the virulence of bacteria. The aim of this study was to investigate the effect of sub-MIC chlorhexidine on biofilm formation and coaggregation of oral early colonizers, such as Streptococcus gordonii, Actinomyces naeslundii and Actinomyces odontolyticus. The biofilm formation of S. gordonii, A. naeslundii and A. odontolyticus was not affected by sub-MIC chlorhexidine. However, the biofilm formation of S. mutans increased after incubation with sub-MIC chlorhexidine. In addition, cell surface hydrophobicity of S. mutans treated with sub-MIC of chlorhexidine, decreased when compared with the group not treated with chlorhexidine. However, significant differences were seen with other bacteria. Coaggregation of A. naeslundii with A. odontolyticus reduced by sub-MIC chlorhexidine, whereas the coaggreagation of A. naeslundii with S. gordonii remained unaffected. These results indicate that sub-MIC chlorhexidine could influence the binding properties, such as biofilm formation, hydrophobicity and coaggregation, in early colonizing streptococci and actinomycetes.

The effect of 4-hexylresorcinol on xenograft degradation in a rat calvarial defect model / 대한악안면성형재건외과학회지

BACKGROUND: The objective of this study was to evaluate xenograft degradation velocity when treated with 4-hexylresorcinol (4HR). METHODS: The scapula of a cow was purchased from a local grocery, and discs (diameter 8 mm, thickness 1 mm) were prepared by trephine bur. Discs treated with 4HR were used as the experimental group. Untreated discs were used as the control. X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), antibacterial test, endotoxin test, and scanning electron microscopy (SEM) were performed on the discs. In vivo degradation was evaluated by the rat calvarial defect model. RESULTS: The XRD and FT-IR results demonstrated successful incorporation of 4HR into the bovine bone. The experimental disc showed antibacterial properties. The endotoxin test yielded results below the level of endotoxin contamination. In the SEM exam, the surface of the experimental group showed needle-shaped crystal and spreading of RAW264.7 cells. In the animal experiments, the amount of residual graft was significantly smaller in the experimental group compared to the control group (P = 0.003). CONCLUSIONS: In this study, 4HR was successfully incorporated into bovine bone, and 4HR-incorporated bovine bone had antibacterial properties. In vivo experiments demonstrated that 4HR-incorporated bovine bone showed more rapid degradation than untreated bovine bone.

Postantibiotic Effects of Photodynamic Therapy Using Erythrosine and Light Emitting Diode on Streptococcus mutans

Dental caries, the most common oral disease, is a multifactorial disease caused by interactions among bacteria within the dental plaque, food, and saliva, resulting in tooth destruction. Streptococcus mutans has been strongly implicated as the causative organism in dental caries and is frequently isolated from human dental plaque. Photodynamic therapy (PDT) is a technique that involves the activation of photosensitizer by light in the presence of tissue oxygen, resulting in the production of reactive radicals capable of inducing cell death. Postantibiotic effect (PAE) is defined as the duration of suppressed bacterial growth following brief exposure to an antibiotic. In this study, the in vitro PAE of PDT using erythrosine and light emitting diode on S. mutans ATCC 25175 was investigated. The PAE of PDT for 1 s irradiation and 3 s irradiation were 1.65 h and 2.1 h, respectively. The present study thus confirmed PAE of PDT using erythrosine on S. mutans.

Effect of fluoride-containing gel on the roughness of a titanium surface and the promotion of bacterial growth

PURPOSE: The aim of this study was to evaluate whether fluorides at various pH cause changes in the surface roughness of titanium implants that alter the adherence of bacterial biofilms. MATERIALS AND METHODS: The titanium disks were assigned randomly to the following seven groups according to the fluoride agents and application time (1 minute or 30 minute) used: control (no treatment); group 1 (1.23% acidulated phosphate fluoride [APF] at pH 3.5 for 1 minute); group 2 (1.23% APF at pH 3.5 for 30 minute); group 3 (1.23% APF at pH 4.0 for 1 minute); group 4 (1.23% APF at pH 4.0 for 30 minute); group 5 (2% NaF gel at pH 7.0 for 1 minute); group 6 (2% NaF gel at pH 7.0 for 30 minute). The surface roughness of the titanium disks and the amount of adherent bacteria were measured. RESULTS: Group 2 showed a significantly greater surface roughness than the control group (P < 0.0001). No significant differences in the amount of surface bacteria were observed between the treated samples and the controls. In addition, there were no significant differences in bacterial adherence relative to the incubation period between the treated samples and the controls. CONCLUSION: The surface roughness of the titanium disks was significantly greater after treatment with APF at pH 3.5 for 30 min compared with that of the controls. In addition, we found that the amount of Porphyromonas gingivalis, Fusobacterium nucleatum, and Aggregatibactor actinomycetemcomitans was similar among all groups.

Effects of Sub Minimal Inhibitory Concentration of Metronidazole and Penicillin on Morphology of Aggregatibacter actinomycetemcomitans: Scanning Electron Microscopy Observation

Minimal inhibitory concentration (MIC) is the lowest concentration of antibiotics that inhibits the visible growth of bacteria. It has been reported that sub-MIC of antibiotics may result in morphological alterations, along with the biochemical and physiological changes in bacteria. The purpose of this study was to examine morphological changes of Aggregatibacter actinomycetemcomitans, after the treatment with sub-MIC metronidazole and penicillin. The bacterial morphology was observed with scanning electron microscope, after incubating with sub-MIC antibiotics. The length of A. actinomycetemcomitans was increased after the incubation with sub-MIC metronidazole and penicillin. Sub-MIC metronidazole and penicillin inhibited bacterial division and induced long filaments. Our study showed that metronidazole and penicillin can induce the morphological changes in A. actinomycetemcomitans.

The effect of photodynamic therapy on Aggregatibacter actinomycetemcomitans attached to surface-modified titanium

PURPOSE: The purpose of this study was to evaluate the effect of photodynamic therapy (PDT) using erythrosine and a green light emitting diode (LED) light source on biofilms of Aggregatibacter actinomycetemcomitans attached to resorbable blasted media (RBM) and sandblasted, large-grit, acid-etched (SLA) titanium surfaces in vitro. METHODS: RBM and SLA disks were subdivided into four groups, including one control group and three test groups (referred to as E0, E30, E60), in order to evaluate the effect of PDT on each surface. The E0 group was put into 500 microL of 20 microM erythrosine for 60 seconds without irradiation, the E30 group was put into erythrosine for 60 seconds and was then irradiated with a LED for 30 seconds, and the E60 group was put into erythrosine for 60 seconds and then irradiated with a LED for 60 seconds. After PDT, sonication was performed in order to detach the bacteria, the plates were incubated under anaerobic conditions on brucella blood agar plates for 72 hours at 37degrees C, and the number of colony-forming units (CFUs) was determined. RESULTS: Significant differences were found between the control group and the E30 and E60 groups (P<0.05). A significantly lower quantity of CFU/mL was found in the E30 and E60 groups on both titanium disk surfaces. In confocal scanning laser microscopy images, increased bacterial death was observed when disks were irradiated for a longer period of time. CONCLUSIONS: These findings suggest that PDT using erythrosine and a green LED is effective in reducing the viability of A. actinomycetemcomitans attached to surface-modified titanium in vitro.

Effect of Sub-Minimal Inhibitory Concentrations of Antibiotics on Biofilm Formation and Coaggregation of Streptococci and Actinomycetes

Minimal inhibitory concentration (MIC) is the lowest antibiotic concentration that inhibits the visible growth of bacteria. Sub-minimal inhibitory concentration (Sub-MIC) is defined as the concentration of an antimicrobial agent that does not have an effect on bacterial growth but can alter bacterial biochemistry, thus reducing bacterial virulence. Many studies have confirmed that sub-MICs of antibiotics can inhibit bacterial virulence factors. However, most studies were focused on Gram-negative bacteria, while few studies on the effect of sub-MICs of antibiotics on Gram-positive bacteria. In this study, we examined the influence of sub-MICs of doxycycline, tetracycline, penicillin and amoxicillin on biofilm formation and coaggregation of Streptococcus gordonii, Streptococcus mutans, Actinomyces naeslundii, and Actinomyces odontolyticus. In this study, incubation with sub-MIC of antibiotics had no effect on the biofilm formation of S. gordonii and A. naeslundii. However, S. mutans showed increased biofilm formation after incubation with sub-MIC amoxicillin and penicillin. Also, the biofilm formation of A. odontolyticus was increased after incubating with sub-MIC penicillin. Coaggregation of A. naeslundii with S. gordonii and A. odontolyticus was diminished by sub-MIC amoxicillin. These observations indicated that sub-MICs of antibiotics could affect variable virulence properties such as biofilm formation and coaggregation in Gram-positive oral bacteria.

Identification of Bacterial Flora on Cellular Phones of Dentists

Dental professionals are repeatedly exposed to many microorganisms present in both blood and saliva. Thus, dental professionals are at a greater risk of acquiring and spreading infections, and the implementation of infections control guidelines is necessary. Cellular phones have become a necessary device for communicating in hospitals. Cellular phones contaminated with bacteria may serve as a fomite in the transmission of pathogens by the hands of medical personnel. Nevertheless, studies about rate and levels of bacterial contamination of cellular phones have been extremely limited with regards to dental personnel. The purpose of this study was to identify bacterial flora on the cellular phones of dentists by a molecular biological method using the 16S rRNA cloning and sequencing method. We acquired total 200 clones from dentists' cell phones and identified the bacterial species. Pseudomonas (34.6%), Lactobacillus (18.5%), Azomonas (11.5%), and Janthinobacterium (6%) were the dominant genera on dentists' cell phones. The oral bacteria identified were Anaerococcus lactolyticus, Gibbsiella dentisursi, Lactobacills leiae, Streptococcus mitis, Streptococcus oligofermentans, and Streptococcus sanguinis. Pathogenic bacteria and opportunistic pathogens such as Carnobacterium funditum, Raoultella planticola, Shigella flexneri, Lactobacillus iners, Staphylococcus aureus, and Staphylococcus epidermidis were also identified.

Incidence of Tetracycline Resistance Genes, tet(M) and tet(O), in Streptococci Isolated from Dental Plaques of Koreans

Streptococci are among the normal human microflora that populate the oral cavity. However, oral streptococci are known as a major causative agent for dental caries and bacterial endocarditis. Tetracycline is a broad-spectrum antibiotic that is used for oral infections but two mechanisms of tetracycline resistance in streptococci have been reported. The tet(K) and tet(L) genes in these bacteria are related to the active efflux of tetracycline, whereas tet(M) and tet(O) confer ribosomal protection from this antibiotic. It has been reported that the tetracycline resistance of streptococci is related mainly to the activity of tet(M) and tet(O). In our present study, we examined the prevalence of tet(M) and tet(O) in oral streptococci isolated from Korean dental plaques using PCR. One hundred and forty eight of 635 isolates (23.3%) were tetracycline resistant; 68 of these strains (46%) harbored tet(M) and 3 strains (2%) were positive for tet(O). However, tet(M) and tet(O) did not co-exist in any of the resistant strains. Seventy seven of the 148 tetracycline resistant strains (52%) were negative for both the tet(M) and tet(O) genes.

Molecular Identification of Bacterial Species Present on Toothbrushes

Toothbrushes play an essential role in oral hygiene. However, they can be significant in microbial transmission and can increase the risk of infection, since they can serve as a reservoir for microorganisms in healthy, oral-diseased and medically ill adults. This study was conducted to evaluate toothbrush contamination in six toothbrushes donated from four people. Two participants each supplied two toothbrushes - one used in the bathroom and one used in the workplace. The other two people each donated two toothbrushes used in the workplace. Polymerase chain reaction was used to construct a 16S rRNA clone library. Sequences of cloned DNA were compared with those from the reference organisms provided by GenBank. A total 120 clones, representing 20 clones for each toothbrush, were analyzed. They are composed of six pylum, 46 genera and 79 species. The most dominant species were Streptococcus oralis, Streptococcus parasanguinis and Haemophilus parainfluenzae. Enterobacter and Escherichia were recovered from toothbrushes used domestically. Toothbrushes used in the workplace did not contain Enterobacteria.

Effect of Sub-Minimal Inhibitory Concentration Antibiotics on Morphology of Periodontal Pathogens

Minimal inhibitory concentration (MIC) is the lowest concentration of antibiotics that inhibits the visible growth of a microorganism. It has been reported that sub-MIC of antibiotics may result in morphological alterations along with biochemical and physiological changes in bacteria. The purpose of this study was to examine morphological changes of periodontal pathogens after treatment with sub-MIC antibiotics. Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Porphyromonas gingivalis were used in this study. The MIC for amoxicillin, doxycycline, metronidazole, penicillin, and tetracycline were determined by broth dilution method. The bacterial morphology was observed with bright field microscope after incubating with sub-MIC antibiotics. The length of A. actinomycetemcomitans and F. nucleatum were increased after incubation with metronidazole; penicillin and amoxicillin. P. gingivalis were increased after incubating with metronidazole and penicillin. However, F. nucleatum showed decreased length after incubation with doxycycline and tetracycline. In this study, we observed that sub-MIC antibiotics can affect the morphology of periodontal pathogens.

Photodynamic bactericidal effect against Enterococcus faecalis by erythrosine concentration and LED irradiation times / 대한구강보건학회지

OBJECTIVES: The purpose of this study was to provide photodynamic bactericidal effect against Enterococcus faecalis by erythrosine concentrations and LED irradiation times. METHODS: Erythrosine was used as a photosensitizer and green LED (3 Watt, 520-530 nm) was used as light source. E. faecalis ATCC 1943 and E. faecalis ATCC 29212 were used in this study. Approximately 10(5) CFU of bacteria were added in wells of a 96-well microtitration plate. For examining the effects of concentrations of erythrosine, 0, 0.625, 1.25, 2.5, 5, and 10 microM of erythrosine were added in wells containing bacteria. The irradiation time with LED was 30 sec. In another set of experiment, the effect of irradiation time for killing of bacteria was investigated by increasing irradiation time from 0 to 30 s with 10 microM of erythrosine final concentration. After irradiation, each sample was serially diluted with PBS and 50 microl of diluents was spread on duplicate blood agar plates. The plates were incubated for 72 h at 37degrees C under aerobic conditions and the number of CFU was determined. The experiments were repeated four times. The results were analyzed using one-way ANOVA, and Tukey's multiple comparison at a significance level of 0.05. RESULTS: When the erythrosine concentrations were more than 2.5 microM, E. faecalis ATCC 29212 was significantly decreased (P<0.05). The more erythrosine concentrations increased, the more E. faecalis ATCC 1943 decreased statistically significantly (P<0.05). In another set of experiment, when LED irradiation time was more than 20 s, E. faecalis ATCC 1943 decreased significantly (P<0.05), and if the irradiation times was more than 5 s, E. faecalis ATCC 29212 decreased significantly (P<0.05). CONCLUSIONS: PDT using erythrosine and green LED was found to be an effective method in killing E. faecalis.