RESUMEN
This study aimed to investigate the photodynamic effects of curcumin, nanomicelle curcumin, and erythrosine on Lactobacillus casei (L. casei). Various concentrations of curcumin (1.5 g/L, 3 g/L), nano-curcumin (3 g/L), and erythrosine (100 µM/L, 250 µM/L) were tested either alone or combined with light irradiation (PDT effect) against L. casei in planktonic and biofilm cultures. The light was emitted from a light-emitting diode (LED) with a central wavelength of 450 nm. A 0.12% chlorhexidine digluconate (CHX) solution served as the positive control, and a solution containing neither photosensitizer nor light was the negative control group. The number of viable microorganisms was determined using serial dilution. There was a significant difference in the viability of L. casei in both planktonic and biofilm forms (P < 0.05). In the planktonic culture, the antibacterial effects of CHX and PDT groups with curcumin 3 g/L and erythrosine 250 µM/L were significantly greater than the other groups (P < 0.05). For L. casei biofilms, the greatest toxic effects were observed in CHX and PDT groups with curcumin 3 g/L, erythrosine 250 µmol/L, erythrosine 100 µmol/L, and nanomicelle curcumin 3 g/L, with a significant difference to other groups (P < 0.05). The antibacterial effects of all photosensitizers (except erythrosine 250 µmol/L at planktonic culture) enhanced significantly when combined with light irradiation (P < 0.05). PDT with curcumin 3 g/L or erythrosine 250 µmol/L produced comparable results to CHX against L. casei at both planktonic and biofilm cultures. Alternatively, PDT with erythrosine 100 µmol/L or nanomicelle curcumin 3 g/L could be suggested to kill L. casei biofilms.
Asunto(s)
Antiinfecciosos , Curcumina , Lacticaseibacillus casei , Fotoquimioterapia , Eritrosina/farmacología , Fotoquimioterapia/métodos , Curcumina/farmacología , Streptococcus mutans/efectos de la radiación , Fármacos Fotosensibilizantes/farmacología , Antiinfecciosos/farmacología , Biopelículas , Antibacterianos/farmacologíaRESUMEN
BACKGROUND: The aim of this study was to evaluate the inactivation potency of riboflavin and curcumin plus blue diode laser against Streptococcus mutans with different power densities. MATERIALS AND METHODS: In this in vitro study, standard-strain S. mutans was exposed to curcumin and riboflavin plus blue diode laser with different power densities (0.4-1.0 W/cm2) as well as chlorhexidine (CHX). The colony forming units (CFUs)/mL was calculated. Data were analyzed by one-way ANOVA. RESULTS: Antibacterial analysis indicated that the blue diode laser irradiation with curcumin and riboflavin provided a satisfactory reduction of the S. mutans level. In addition, S. mutans was more affected by curcumin + blue diode laser when the power density was set to 1.0 W/cm2 (P < 0.0001). Meanwhile, bacterial suspensions treated with CHX showed maximum colony number reduction, compared with the control (P < 0.0001). CONCLUSION: This study showed the blue diode laser along with curcumin had strong bactericidal effect on S. mutans, and this effect improved by increasing the power density.
Asunto(s)
Antiinfecciosos , Curcumina , Fotoquimioterapia , Fotoquimioterapia/métodos , Streptococcus mutans/efectos de la radiación , Fármacos Fotosensibilizantes/farmacología , Curcumina/farmacología , Clorhexidina/farmacología , Antibacterianos , Riboflavina , BiopelículasRESUMEN
The direct application of light for photo-disinfection potentially provides a safe and novel modality to inhibit or eliminate cariogenic bacteria residing upon and within dentine. This study aimed to both; characterize the pattern of transmission of 405 nm light through molar dentine at different tooth locations, as well as, determine the irradiation parameters that are antibacterial for Streptococcus mutans under various growth conditions, including lawns, planktonic cultures, and biofilms. To determine the amount of light (405 nm) transmitted at different anatomical tooth locations; irradiance values were recorded after blue light (470-4054 mW/cm2) had traversed through occlusal, oblique, and buccal dentine sections; and three thicknesses - 1, 2 and 3 mm were investigated. To determine tubular density; scanning electron micrographs from 2 mm outer (dentine-enamel junction) and inner (pulp) dentine sections were analysed. For photo-disinfection studies; S. mutans was irradiated using the same 405 nm wavelength light at a range of doses (110-1254 J/cm2) in both biofilm and planktonic cultures. The inhibitory effect of the irradiation on bacterial lawns was compared by measuring zones of inhibition; and for planktonic cultures both spectrophotometric and colony forming unit (CFU) assays were performed. A live/dead staining assay was utilised to determine the effect of irradiation on bacterial viability in mature biofilms. Data indicated that increasing dentine thickness decreased light transmission significantly irrespective of its orientation. Occlusal and oblique samples exhibited higher transmission compared with buccal dentine. Oblique dentine 405 nm light transmission was comparable with that of occlusal dentine independent of section thickness. An increased tubule density directly positively correlated with light transmission. Irradiation at 405 nm inhibited S. mutans growth in both biofilm and planktonic cultures and a dose response relationship was observed. Irradiation at doses of 340 and 831 J/cm2 led to significant reductions in bacterial growth and viability; as determined by CFU counting and live/dead staining. Data suggests that phototherapy approaches utilising a 405 nm wavelength have therapeutic potential to limit cariogenic bacterial infections both at the surface and within dentine.
Asunto(s)
Dentina/efectos de la radiación , Desinfección/métodos , Luz , Adulto , Color , Dentina/microbiología , Femenino , Humanos , Masculino , Viabilidad Microbiana/efectos de la radiación , Streptococcus mutans/fisiología , Streptococcus mutans/efectos de la radiación , Adulto JovenRESUMEN
OBJECTIVE: We evaluated the effect of antimicrobial photodynamic therapy (a-PDT) with Rose Bengal and blue light LED on bacteria that initiate and promote dental caries. DESIGN: Colony forming units of Streptococcus mutans, Streptococcus sobrinus, Streptococcus sanguinis, and Lactobacillus salivarius under planktonic and biofilm conditions were counted after a-PDT treatment using Rose Bengal and blue light LED. In addition, cariogenic bacteria from saliva and dental plaques from ten volunteers were used for evaluation of a-PDT treatment. RESULTS: We found that a-PDT using Rose Bengal at > 10 µg/mL had antimicrobial effects on oral Gram-positive S. mutans, S. sobrinus, S. sanguinis, and L. salivarius under both planktonic and biofilm conditions. The effect was also observed for cariogenic bacteria that formed biofilms containing water-insoluble glucans, through which the bacteria are firmly attached to the tooth surface. Moreover, a-PDT led to a marked reduction in cariogenic bacteria in saliva and dental plaques. CONCLUSION: a-PDT could be a useful approach for controlling dental caries in dental surgery.
Asunto(s)
Antiinfecciosos , Fotoquimioterapia , Rosa Bengala/farmacología , Streptococcus mutans/efectos de los fármacos , Streptococcus mutans/efectos de la radiación , Antiinfecciosos/farmacología , Biopelículas/efectos de los fármacos , Biopelículas/efectos de la radiación , Caries Dental/tratamiento farmacológico , HumanosRESUMEN
Caries-related biofilms and associated complications are significant threats in dentistry, especially when biofilms grow over dental restorations. The inhibition of cariogenic biofilm associated with the onset of carious lesions is crucial for preventing disease recurrence after treatment. This in vitro study defined optimized parameters for using a photosensitizer, toluidine blue O (TBO), activated via a red light-emitting diode (LED)-based wireless device to control the growth of cariogenic biofilms. The effect of TBO concentrations (50, 100, 150, and 200 µg/mL) exposed to light or incubated in the dark was investigated in successive cytotoxicity assays. Then, a mature Streptococcus mutans biofilm model under sucrose challenge was treated with different TBO concentrations (50, 100, and 150 µg/mL), different light energy doses (36, 108, and 180 J/cm2), and different incubation times before irradiation (1, 3, and 5 min). The untreated biofilm, irradiation with no TBO, and TBO incubation with no activation represented the controls. After treatments, biofilms were analyzed via S. mutans colony-forming units (CFUs) and live/dead assay. The percentage of cell viability was within the normal range compared to the control when 50 and 100 µg/mL of TBO were used. Increasing the TBO concentration and energy dose was associated with biofilm inhibition (p < 0.001), while increasing incubation time did not contribute to bacterial elimination (p > 0.05). Irradiating the S. mutans biofilm via 100 µg/mL of TBO and ≈180 J/cm2 energy dose resulted in ≈3-log reduction and a higher amount of dead/compromised S. mutans colonies in live/dead assay compared to the control (p < 0.001). The light energy dose and TBO concentration optimized the bacterial elimination of S. mutans biofilms. These results provide a perspective on the determining parameters for highly effective photo-killing of caries-related biofilms and display the limitations imposed by the toxicity of the antibacterial photodynamic therapy's chemical components. Future studies should support investigations on new approaches to improve or overcome the constraints of opportunities offered by photodynamic inactivation of caries-related biofilms.
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Biopelículas/efectos de la radiación , Luces de Curación Dental , Caries Dental/terapia , Streptococcus mutans/efectos de la radiación , Animales , Recuento de Colonia Microbiana , Caries Dental/microbiología , Relación Dosis-Respuesta en la Radiación , Ratones , Fármacos Fotosensibilizantes/efectos adversos , Células RAW 264.7 , Streptococcus mutans/patogenicidad , Streptococcus mutans/fisiología , Cloruro de Tolonio/efectos adversosRESUMEN
Dental caries is a complex multifactorial chronic infectious disease guided by several risk or protective factors. Saliva has an important role in caries and the remineralization process. Caries risk assessment is defined as the probability of new caries lesion development or the existing lesion progression in a given time period. Caries diagnostics and risk factor assessment are followed by targeted elimination of risk factors and less conservative but abundant preventive therapeutic measures. The aim of our prospective randomized study was to elucidate on how photobiomodulation of major salivary glands with polychromatic light or LED light affects caries risk factors in high caries-risk patients. Thirty-six patients were assigned to one of the following three experimental groups: the first, irradiated with polarized polychromatic light (40 mW/cm2, wavelengths 480-3400 nm); the second, a continuous LED light (16 mW/cm2, wavelengths 625, 660, 850 nm); the third, same LED light in a pulsed mode. The fourth group was the control, for which a non-therapeutic visible light was used. Light was administered extra-orally bilaterally above the parotid and submandibular glands for 10 min and intra-orally above the sublingual glands for 5 min, 3 times a week, for 4 consecutive weeks. Each patient's caries risk was assessed according to Cariogram before and after therapy. Caries risk factors were determined from samples of saliva before therapy, two weeks after it commenced, at the end of therapy, and four weeks after the end of therapy. At the end of treatment, the following findings were obtained: In the group irradiated with polarized polychromatic light and in the group irradiated with continuous LED light, the Streptococcus mutans and Lactobacillus counts decreased and salivary buffering capacity increased (p < 0.05). In the group irradiated with pulsed LED light, Streptococcus mutans counts decreased and unstimulated salivary flow and salivary buffering capacity increased (p < 0.05). In all three experimental groups, caries risk was lower (p < 0.05). In the placebo control group, there were no statistically significant differences between parameters before and after therapy. We concluded that photobiomodulation of major salivary glands in high caries-risk patients can reduce the cariogenic bacteria in saliva and improve some salivary parameters, thus reducing caries risk.
Asunto(s)
Caries Dental/microbiología , Caries Dental/prevención & control , Terapia por Luz de Baja Intensidad , Glándulas Salivales/microbiología , Glándulas Salivales/efectos de la radiación , Carga Bacteriana/efectos de la radiación , Femenino , Humanos , Lactobacillus/fisiología , Lactobacillus/efectos de la radiación , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Medición de Riesgo , Streptococcus mutans/fisiología , Streptococcus mutans/efectos de la radiaciónRESUMEN
Objective: This study was performed to determine the bactericidal effects of erbium:yttrium-aluminum-garnet (Er:YAG) laser irradiation and the morphological and chemical composition changes in bovine dentin. Methods: Dentin slabs were prepared from bovine incisors, and then cultured with Streptococcus mutans to produce bacteria-infected dentin samples. The samples were randomly divided into five groups with Er:YAG laser irradiation energy densities of 0, 6.37, 12.73, 19.11, and 25.47 J/cm2. After irradiation, samples were stained and observed by confocal laser scanning microscopy. The bactericidal abilities were measured using live/dead staining. The morphology and chemical components were investigated by scanning electron microscopy and energy-dispersive spectrometry. Results: After irradiation, the elimination of bacteria and the smear layer were significantly better in the high energy density groups (19.11, 25.47 J/cm2) than in the low energy density groups (6.37, 12.73 J/cm2; p < 0.001). On morphological examination, the group with minimum energy density (6.37 J/cm2) showed superficial melting. In the high energy density groups (12.73, 19.11, and 25.47 J/cm2), laser-irradiated dentin showed a clean surface with open orifices. Significant increases were observed in the weight percentages of calcium (from 19.75 ± 0.69 to 34.47 ± 2.91, p < 0.001) and phosphate (from 8.58 ± 0.43 to 15.10 ± 1.81, p < 0.001), whereas significant decreases were observed for oxygen (from 49.84 ± 0.69 to 36.39 ± 2.86, p < 0.001) and carbon (from 26.06 ± 3.58 to 12.80 ± 2.26, p < 0.01) with increasing energy density. Conclusions: This study confirmed that Er:YAG laser irradiation has bactericidal and dentin conditioning effects.
Asunto(s)
Dentina/microbiología , Dentina/efectos de la radiación , Láseres de Estado Sólido/uso terapéutico , Terapia por Luz de Baja Intensidad , Streptococcus mutans/efectos de la radiación , Animales , Bovinos , Dentina/diagnóstico por imagen , Microscopía Confocal , Técnicas de Cultivo de TejidosRESUMEN
The aim of this in vitro study was to determine the effect of violet-blue light on the metabolic activity of early Streptococcus mutans biofilm, reincubated at 0, 2, and 6 h after 5 min of violet-blue light treatment. S. mutans UA159 biofilm cells were cultured for 12 to 16 h in microtiter plates with Tryptic Soy broth (TSB) or TSB with 1% sucrose (TSBS) and irradiated with violet-blue light for 5 min. After irradiation, the plates were reincubated at 37°C for 0, 2, or 6 h in 5% CO2. Colorimetric tetrazolium salt reduction assay was used to investigate bacterial metabolic activity. Mixed model ANOVA was used to find the difference between the violet-blue light treated and nontreated groups. Bacterial metabolic activity was significantly lower in the violet-blue light group for TSB than in the nontreated group (P < 0.0001) regardless of recovery time. However, the differences between metabolic activity in the treated groups without sucrose decreased over time. For TSBS, metabolic activity was significantly lower with violet-blue light at 0 and 2 h. Violet-blue light inhibited the metabolic activity of S. mutans biofilm cells in the light-treated group. This finding may present a unique treatment method for patients with active caries.
Asunto(s)
Biopelículas , Colorimetría/métodos , Fototerapia , Streptococcus mutans/metabolismo , Streptococcus mutans/efectos de la radiación , Sales de Tetrazolio/química , HumanosRESUMEN
The aim of the present study was to evaluate, in vitro, the effect of different pre-irradiation times of the photosensitizer in photodynamic therapy in biofilms formed by Streptococcus mutans and Candida albicans, through the evaluation of the microbial load. The factors under study were as follows: times of pre-irradiation of the photosensitizer in three levels (1, 2, or 5 min). For the control of the cariogenic dental biofilm with antimicrobial photodynamic therapy (aPDT), methylene blue (0.01%) was used in association with the diode laser (InGaAlP) with a wavelength of 660 nm. Chlorhexidine digluconate (0.12% CHX) and saline were used as positive and negative controls, respectively. The study design was carried out in complete and randomized blocks. The sample consisted of 15 S. mutans biofilms cultures, randomly divided into five groups and 15 C. albicans cultures, also divided into five groups. The experiment was performed in triplicate (n = 3) and the response variables were obtained through quantitative analysis of bacterial viability, expressed in colony-forming units (CFU) per square millimeter of the specimen area. The data were analyzed with the aid of the ANOVA one-way test and Tukey's post-test. All analyses were performed using the Graph Pad Prism 4.0 program, with a significance level of 5%. For the S. mutans group, only the saline solution presented a statistically significant difference when compared to the other treatments (p < 0.05), that is, the treatment with aPDT, irrespective of the irradiation time applied, was similar to the treatment with CHX and both were more effective in reducing cariogenic biofilm compared to saline. For the group of C. albicans, there was no statistical difference between the groups (p > 0.05). Therefore, it can be concluded that the treatment with aPDT reduced the number of CFUs of S. mutans in a similar way to CHX, independently of the pre-irradiation time applied. No effect of this therapy or of the different pre-irradiation times on the C. albicans biofilm could be observed. In this way, the pre-irradiation time of 1 min can be used to reduce the microbial load of S. mutans.
Asunto(s)
Antibacterianos/farmacología , Láseres de Semiconductores , Fotoquimioterapia , Biopelículas/efectos de los fármacos , Candida albicans/efectos de los fármacos , Candida albicans/crecimiento & desarrollo , Candida albicans/efectos de la radiación , Clorhexidina/análogos & derivados , Clorhexidina/farmacología , Recuento de Colonia Microbiana , Humanos , Azul de Metileno/farmacología , Pruebas de Sensibilidad Microbiana , Viabilidad Microbiana/efectos de los fármacos , Viabilidad Microbiana/efectos de la radiación , Fármacos Fotosensibilizantes/farmacología , Streptococcus mutans/efectos de los fármacos , Streptococcus mutans/crecimiento & desarrollo , Streptococcus mutans/efectos de la radiación , Factores de TiempoRESUMEN
BACKGROUND: Ultraviolet (UV) light is used for phototherapy in dermatology, and UVB light (around 310 nm) is effective for treatment of psoriasis and atopic dermatitis. In addition, it is known that UVC light (around 265 nm) has a bactericidal effect, but little is known about the bactericidal effect of UVB light. In this study, we examined the bactericidal effects of UVB-light emitting diode (LED) irradiation on oral bacteria to explore the possibility of using a 310 nm UVB-LED irradiation device for treatment of oral infectious diseases. METHODS: We prepared a UVB (310 nm) LED device for intraoral use to examine bactericidal effects on Streptococcus mutans, Streptococcus sauguinis, Porphyromonas gingivalis, and Fusobacterium nucleatum and also to examine the cytotoxicity to a human oral epithelial cell line (Ca9-22). We also examined the production of nitric oxide and hydrogen peroxide from Ca9-22 cells after irradiation with UVB-LED light. RESULTS: Irradiation with the 310 nm UVB-LED at 105 mJ/cm2 showed 30-50% bactericidal activity to oral bacteria, though 17.1 mJ/cm2 irradiation with the 265 nm UVC-LED completely killed the bacteria. Ca9-22 cells were strongly injured by irradiation with the 265 nm UVC-LED but were not harmed by irradiation with the 310 nm UVB-LED. Nitric oxide and hydrogen peroxide were produced by Ca9-22 cells with irradiation using the 310 nm UVB-LED. P. gingivalis was killed by applying small amounts of those reactive oxygen species (ROS) in culture, but other bacteria showed low sensitivity to the ROS. CONCLUSIONS: Narrowband UVB-LED irradiation exhibited a weak bactericidal effect on oral bacteria but showed low toxicity to gingival epithelial cells. Its irradiation also induces the production of ROS from oral epithelial cells and may enhance bactericidal activity to specific periodontopathic bacteria. It may be useful as a new adjunctive therapy for periodontitis.
Asunto(s)
Fusobacterium nucleatum/efectos de la radiación , Porphyromonas gingivalis/efectos de la radiación , Streptococcus mutans/efectos de la radiación , Streptococcus/efectos de la radiación , Rayos Ultravioleta , Células Epiteliales/citología , Células Epiteliales/metabolismo , Células Epiteliales/efectos de la radiación , Humanos , Peróxido de Hidrógeno/metabolismo , Mucosa Bucal/citología , Mucosa Bucal/metabolismo , Mucosa Bucal/efectos de la radiación , Óxido Nítrico/metabolismo , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Among various preventive approaches, non-invasive phototherapy/photodynamic therapy is one of the methods used to control oral biofilm. Studies indicate that light at specific wavelengths has a potent antibacterial effect. The objective of this study was to determine the effectiveness of violet-blue light at 380-440 nm to inhibit biofilm formation of Streptococcus mutans or kill S. mutans. S. mutans UA159 biofilm cells were grown for 12-16 h in 96-well flat-bottom microtiter plates using tryptic soy broth (TSB) or TSB with 1 % sucrose (TSBS). Biofilm was irradiated with violet-blue light for 5 min. After exposure, plates were re-incubated at 37 °C for either 2 or 6 h to allow the bacteria to recover. A crystal violet biofilm assay was used to determine relative densities of the biofilm cells grown in TSB, but not in TSBS, exposed to violet-blue light. The results indicated a statistically significant (P < 0.05) decrease compared to the non-treated groups after the 2 or 6 h recovery period. Growth rates of planktonic and biofilm cells indicated a significant reduction in the growth rate of the violet-blue light-treated groups grown in TSB and TSBS. Biofilm viability assays confirmed a statistically significant difference between violet-blue light-treated and non-treated groups in TSB and TSBS. Visible violet-blue light of the electromagnetic spectrum has the ability to inhibit S. mutans growth and reduce the formation of S. mutans biofilm. This in vitro study demonstrated that violet-blue light has the capacity to inhibit S. mutans biofilm formation. Potential clinical applications of light therapy in the future remain bright in preventing the development and progression of dental caries.
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Biopelículas/efectos de la radiación , Streptococcus mutans/fisiología , Streptococcus mutans/efectos de la radiación , Luz , Streptococcus mutans/genética , Streptococcus mutans/crecimiento & desarrolloRESUMEN
STATEMENT OF PROBLEM: Microwave irradiation and immersion in solutions have been recommended for denture disinfection. However, the effect of dry conditions and impression materials has not been completely evaluated. PURPOSE: The purpose of this study was to evaluate the effectiveness of microwave irradiation and hydrogen peroxide for the disinfection of dental impression materials. MATERIAL AND METHODS: Specimens (diameter 10 mm, thickness 2 mm) were made with polyvinyl siloxane. Experimental groups were treated with hydrogen peroxide (group H), microwave irradiation (group M), and a combination of both hydrogen peroxide and microwave irradiation (group MH) for 1 minute, 2 minutes, and 3 minutes. The control group was untreated. The total sample size was 120. The specimens were divided into 2 groups, those exposed to Streptococcus mutans and those exposed to Escherichia coli. The disinfection effect and physical properties (contact angle, compatibility with gypsum, strain in compression, tear strength) were evaluated. RESULTS: All 3 groups (H, M, MH) were effective in reducing the number of colony forming units (CFU) per unit volume (mL) for both S mutans and E coli compared with the control. The most significant reduction in the CFU/mL of both bacteria was noted in the MH group and was used to compare either treatment alone (P<.05). No statistically significant difference was noted between the control and treatment groups in terms of all of the physical properties tested (P>.05). CONCLUSIONS: Microwave irradiation was identified as a useful disinfection method against S mutans and E coli, especially when combined with H2O2, without adversely affecting the physical properties of dental impression materials.
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Desinfectantes Dentales/uso terapéutico , Materiales de Impresión Dental/química , Desinfección/métodos , Peróxido de Hidrógeno/uso terapéutico , Microondas/uso terapéutico , Carga Bacteriana/efectos de los fármacos , Carga Bacteriana/efectos de la radiación , Técnicas Bacteriológicas , Sulfato de Calcio/química , Materiales de Impresión Dental/efectos de la radiación , Análisis del Estrés Dental/instrumentación , Escherichia coli/efectos de los fármacos , Escherichia coli/efectos de la radiación , Humanos , Ensayo de Materiales , Polivinilos/química , Polivinilos/efectos de la radiación , Dosis de Radiación , Siloxanos/química , Siloxanos/efectos de la radiación , Streptococcus mutans/efectos de los fármacos , Streptococcus mutans/efectos de la radiación , Estrés Mecánico , Propiedades de Superficie , Temperatura , Resistencia a la Tracción , Factores de Tiempo , HumectabilidadRESUMEN
BACKGROUND: Increasing resistance of oral pathogens to conventional antibacterial agents has resulted to find alternative therapies to overcome resistance development problems; hence this in vitro study was carried out to investigate the efficacy of photoelimination of Streptococcus mutans with two methods of photodynamic and photothermal therapy. METHODS: Standard Suspensions of S. mutans were treated in two groups of photodynamic therapy with Toluidine blue O and Rhadachlorin(®) and photothermal therapy by EmunDo(®) and their individual light sources, then Bacterial suspension from each treatment was subcultured on the surface of Mueller-Hinton agar plates and bacterial growth was assessed. The results were analyzed by analysis of variance and Tukey test (p<0.05). RESULTS: After treatments significant reduction of S. mutans viability in planktonic culture was observed in both groups of photodynamic and photothermal therapy with no priority. CONCLUSION: Photoelimination can be a novel modality in the eradication of S. mutans colonies in near future.
Asunto(s)
Desinfección/métodos , Fotoquimioterapia/métodos , Fototerapia/métodos , Streptococcus mutans/efectos de los fármacos , Streptococcus mutans/efectos de la radiación , Cloruro de Tolonio/administración & dosificación , Apoptosis/efectos de los fármacos , Apoptosis/fisiología , Apoptosis/efectos de la radiación , Terapia Combinada/métodos , Fármacos Fotosensibilizantes/administración & dosificación , Streptococcus mutans/fisiologíaRESUMEN
Conventional antibacterial treatment fails to eradicate biofilms associated with common infections of the oral cavity. Unlike chemical agents, which are less effective than anticipated, owing to diffusion limitations in biofilms, light is more effective on bacteria in biofilm than in suspension. Effectiveness depends also on the type and parameters of the light. We tested the phototoxic effects of non-coherent blue light (wavelengths, 400-500 nm) and CO(2) laser (wavelength, 10.6 µm), which have different mechanisms of action on the oral bacterium Streptoccocus mutans, in biofilm and on tooth enamel. Exposure of S. mutans in biofilm to blue light had a delayed effect on bacterial viability throughout the biofilm and a sustained antibacterial effect on biofilm newly formed by previously irradiated bacteria. A synergistic antibacterial effect between blue light and H(2)O(2) may enhance the phototoxic effect, which involves a photochemical mechanism mediated by reactive oxygen species (ROS) formation. The effect of CO(2) laser irradiation on the viability of S. mutans in biofilm on enamel samples appeared to be higher in the deep layers, due to heating of the enamel surface by the absorbed energy. Biofilms do not interfere with the chemical changes resulting from irradiation, which may increase the enamel's resistance to acid attack.
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Biopelículas/efectos de la radiación , Esmalte Dental/efectos de la radiación , Placa Dental/radioterapia , Láseres de Gas/uso terapéutico , Terapia por Luz de Baja Intensidad , Fototerapia/métodos , Streptococcus mutans/efectos de la radiación , Humanos , Peróxido de Hidrógeno/farmacología , Láseres de Gas/efectos adversos , Viabilidad Microbiana/efectos de la radiaciónRESUMEN
UNLABELLED: Several photosensitizers have been used against oral bacteria without standardization. Singlet oxygen ((1)O(2)) is an aggressive chemical species that can kill cells through apoptosis or necrosis. OBJECTIVE: to compare the antimicrobial activity of photodynamic therapy (PDT) with different photosensitizers at the same concentration against Streptococcus mutans. In addition, the (1)O(2) production of each photosensitizer was determined. The photosensitizers (163.5 µM) methylene blue (MB), toluidine blue ortho (TBO) and malachite green (MG) were activated with a light-emitting diode (LED; λ=636 nm), while eosin (EOS), erythrosine (ERI) and rose bengal (RB) were irradiated with a curing light (λ=570 nm). Light sources were operated at 24 J cm(-2). For each photosensitizer, 40 randomized assays (n=10 per condition) were performed under one of the following experimental conditions: no light irradiation or photosensitizer, irradiation only, photosensitizer only or irradiation in the presence of a photosensitizer. After treatment, serial dilutions of S. mutans were seeded onto brain heart infusion agar to determine viability in colony-forming units per milliliter (CFU mL(-1)). Generation of (1)O(2) was analyzed by tryptophan photooxidation, and the decay constant was estimated. Results were analyzed by one-way ANOVA and the Tukey-Kramer test (p<0.05). PDT with irradiation in the presence of the photosensitizers TBO and MG was effective in reducing S. mutans counts by 3 and 1.4 logs, respectively (p<0.01), compared to their respective untreated controls. MB generated 1.3 times more (1)O(2) than TBO, and both produced significantly higher concentrations of singlet oxygen than the other photosensitizers. Since in vitro bulk (1)O(2) production does not indicate that (1)O(2) was generated in the bacterial activity site, the bactericidal action against S. mutans cannot be related to in vitro singlet O(2) generation rate. In vitroS. mutans-experiments demonstrated TBO as the only photosensitizer that effectively reduced 99.9% of these microorganisms.
Asunto(s)
Antiinfecciosos/farmacología , Fármacos Fotosensibilizantes/farmacología , Streptococcus mutans/efectos de los fármacos , Antiinfecciosos/química , Eosina Amarillenta-(YS)/química , Eritrosina/química , Luz , Azul de Metileno/química , Azul de Metileno/farmacología , Oxidación-Reducción , Fármacos Fotosensibilizantes/química , Colorantes de Rosanilina/química , Colorantes de Rosanilina/farmacología , Rosa Bengala/química , Oxígeno Singlete/metabolismo , Streptococcus mutans/efectos de la radiación , Cloruro de Tolonio/química , Cloruro de Tolonio/farmacología , Triptófano/químicaRESUMEN
The aim of this study was to evaluate the effect of specific parameters of low-level laser therapy (LLLT) on biofilms formed by Streptococcus mutans, Candida albicans or an association of both species. Single and dual-species biofilms--SSB and DSB--were exposed to laser doses of 5, 10 or 20 J/cm(2) from a near infrared InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm, 0.04 W). After irradiation, the analysis of biobilm viability (MTT assay), biofilm growth (cfu/mL) and cell morphology (SEM) showed that LLLT reduced cell viability as well as the growth of biofilms. The response of S. mutans (SSB) to irradiation was similar for all laser doses and the biofilm growth was dose dependent. However, when associated with C. albicans (DSB), S. mutans was resistant to LLLT. For C. albicans, the association with S. mutans (DSB) caused a significant decrease in biofilm growth in a dose-dependent fashion. The morphology of the microorganisms in the SSB was not altered by LLLT, while the association of microbial species (DSB) promoted a reduction in the formation of C. albicans hyphae. LLLT had an inhibitory effect on the microorganisms, and this capacity can be altered according to the interactions between different microbial species.
Asunto(s)
Biopelículas/efectos de la radiación , Candida albicans/efectos de la radiación , Láseres de Semiconductores , Terapia por Luz de Baja Intensidad/instrumentación , Boca/microbiología , Streptococcus mutans/efectos de la radiación , Técnicas Bacteriológicas , Biopelículas/crecimiento & desarrollo , Candida albicans/crecimiento & desarrollo , Candida albicans/ultraestructura , Colorantes , Relación Dosis-Respuesta en la Radiación , Humanos , Hifa/efectos de la radiación , Ensayo de Materiales , Interacciones Microbianas/efectos de la radiación , Viabilidad Microbiana/efectos de la radiación , Microscopía Electrónica de Rastreo , Micología/métodos , Dosis de Radiación , Streptococcus mutans/crecimiento & desarrollo , Streptococcus mutans/ultraestructura , Succinato Deshidrogenasa/análisis , Temperatura , Sales de Tetrazolio , Tiazoles , Factores de TiempoRESUMEN
The aim of this study was to evaluate the effect of specific parameters of low-level laser therapy (LLLT) on biofilms formed by Streptococcus mutans, Candida albicans or an association of both species. Single and dual-species biofilms - SSB and DSB - were exposed to laser doses of 5, 10 or 20 J/cm2 from a near infrared InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm, 0.04 W). After irradiation, the analysis of biobilm viability (MTT assay), biofilm growth (cfu/mL) and cell morphology (SEM) showed that LLLT reduced cell viability as well as the growth of biofilms. The response of S. mutans (SSB) to irradiation was similar for all laser doses and the biofilm growth was dose dependent. However, when associated with C. albicans (DSB), S. mutans was resistant to LLLT. For C. albicans, the association with S. mutans (DSB) caused a significant decrease in biofilm growth in a dose-dependent fashion. The morphology of the microorganisms in the SSB was not altered by LLLT, while the association of microbial species (DSB) promoted a reduction in the formation of C. albicans hyphae. LLLT had an inhibitory effect on the microorganisms, and this capacity can be altered according to the interactions between different microbial species.
O objetivo deste estudo foi avaliar o efeito de parâmetros específicos de irradiação com laser de baixa intensidade sobre biofilmes formados por Streptococcus mutans (S. mutans), Candida albicans (C. albicans) ou associação de ambas as espécies. Biofilmes isolados ou associados destes microrganismos foram irradiados com um dispositivo laser infra-vermelho próximo de diodos InGaAsP (LaserTABLE 780 ±3 nm, 0,04W), utilizando-se para isto o dispositivo LASERTable. Quinze horas após a irradiação, foi demonstrado, por meio da avaliação da viabilidade celular (Teste de MTT), da morfologia das células (MEV) e do crescimento do biofilme (UFC/mL), que esta terapia foi capaz de reduzir o metabolismo celular, número de microrganismos presentes no biofilme, bem como seu crescimento no local. Quanto à viabilidade celular, a resposta à irradiação do biofilme de S. mutans (SSB) foi semelhante para todas as doses de energia, sendo que o crescimento do biofilme foi dose dependente. Porém, quando associado à C. albicans, este microrganismo apresentou resistência à fototerapia. Já a C. albicans associada ao S. mutans apresentou redução de crescimento significativa, sendo este resultado também foi dose dependente. A morfologia dos microrganismos não foi alterada pelas irradiações realizadas quando em biofilmes isolados. A associação entre os microrganismos promoveu redução na formação de hifas pela C. albicans. A laserterapia de baixa intensidade apresentou efeito inibitório sobre microrganismos, sendo que esta capacidade pode ser alterada de acordo com a interação entre diferentes microrganismos.
Asunto(s)
Humanos , Biopelículas/efectos de la radiación , Candida albicans/efectos de la radiación , Láseres de Semiconductores , Terapia por Luz de Baja Intensidad/instrumentación , Boca/microbiología , Streptococcus mutans/efectos de la radiación , Técnicas Bacteriológicas , Biopelículas/crecimiento & desarrollo , Candida albicans/crecimiento & desarrollo , Candida albicans/ultraestructura , Colorantes , Relación Dosis-Respuesta en la Radiación , Hifa/efectos de la radiación , Ensayo de Materiales , Microscopía Electrónica de Rastreo , Interacciones Microbianas/efectos de la radiación , Viabilidad Microbiana/efectos de la radiación , Micología/métodos , Dosis de Radiación , Streptococcus mutans/crecimiento & desarrollo , Streptococcus mutans/ultraestructura , Succinato Deshidrogenasa/análisis , Temperatura , Factores de Tiempo , Sales de Tetrazolio , TiazolesRESUMEN
The objective of this study was to evaluate the effect of photodynamic therapy with erythrosine and rose bengal using a light-emitting diode (LED) on planktonic cultures of S. mutans. Ten S. mutans strains, including nine clinical strains and one reference strain (ATCC 35688), were used. Suspensions containing 106 cells/mL were prepared for each strain and were tested under different experimental conditions: a) LED irradiation in the presence of rose bengal as a photosensitizer (RB+L+); b) LED irradiation in the presence of erythrosine as a photosensitizer (E+L+); c) LED irradiation only (P-L+); d) treatment with rose bengal only (RB+L-); e) treatment with erythrosine only (E+L-); and f) no LED irradiation or photosensitizer treatment, which served as a control group (P-L-). After treatment, the strains were seeded onto BHI agar for determination of the number of colony-forming units (CFU/mL). The results were submitted to analysis of variance and the Tukey test (p ≤ 0.05). The number of CFU/mL was significantly lower in the groups submitted to photodynamic therapy (RB+L+ and E+L+) compared to control (P-L-), with a reduction of 6.86 log10 in the RB+L+ group and of 5.16 log10 in the E+L+ group. Photodynamic therapy with rose bengal and erythrosine exerted an antimicrobial effect on all S. mutans strains studied.
Asunto(s)
Caries Dental/tratamiento farmacológico , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/farmacología , Streptococcus mutans/efectos de los fármacos , Análisis de Varianza , Carga Bacteriana , Biopelículas/efectos de los fármacos , Biopelículas/efectos de la radiación , Células Cultivadas , Eritrosina/farmacología , Rosa Bengala/farmacología , Streptococcus mutans/aislamiento & purificación , Streptococcus mutans/efectos de la radiación , Factores de TiempoRESUMEN
The objective of this study was to evaluate the effect of photodynamic therapy with erythrosine and rose bengal using a light-emitting diode (LED) on planktonic cultures of S. mutans. Ten S. mutans strains, including nine clinical strains and one reference strain (ATCC 35688), were used. Suspensions containing 10(6) cells/mL were prepared for each strain and were tested under different experimental conditions: a) LED irradiation in the presence of rose bengal as a photosensitizer (RB+L+); b) LED irradiation in the presence of erythrosine as a photosensitizer (E+L+); c) LED irradiation only (P-L+); d) treatment with rose bengal only (RB+L-); e) treatment with erythrosine only (E+L-); and f) no LED irradiation or photosensitizer treatment, which served as a control group (P-L-). After treatment, the strains were seeded onto BHI agar for determination of the number of colony-forming units (CFU/mL). The results were submitted to analysis of variance and the Tukey test (p < 0.05). The number of CFU/mL was significantly lower in the groups submitted to photodynamic therapy (RB+L+ and E+L+) compared to control (P-L-), with a reduction of 6.86 log10 in the RB+L+ group and of 5.16 log10 in the E+L+ group. Photodynamic therapy with rose bengal and erythrosine exerted an antimicrobial effect on all S. mutans strains studied.
Asunto(s)
Caries Dental/tratamiento farmacológico , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/farmacología , Streptococcus mutans/efectos de los fármacos , Análisis de Varianza , Carga Bacteriana , Biopelículas/efectos de los fármacos , Biopelículas/efectos de la radiación , Células Cultivadas , Eritrosina/farmacología , Rosa Bengala/farmacología , Streptococcus mutans/aislamiento & purificación , Streptococcus mutans/efectos de la radiación , Factores de TiempoRESUMEN
UNLABELLED: This study was conducted in order to demonstrate the bactericidal effect of an erbium laser (Er,Cr:YSGG) on Streptococcus mutans, in vivo, at different laser power settings, obtaining sterile enamel surfaces with minimal tissue ablation. The practicians use the laser for the cutting effect on hard tissue to eliminate all the infected and affected enamel and dentine, without collateral damage on the pulp and for its bactericidal effect on cariogenic bacteria. MATERIAL AND METHOD: The study group included 132 superficial carious lesions, on 66 patients with ages between 6 and 20; a two-year study (January 2006 - November 2008) performed in a private practice. The carious lesions were ablated with an erbium laser with two types of tips. A sample of occlusal plaque was taken using a sterile swab, soaked in normal saline solution (before and after the preparation). RESULTS: Before the laser treatment, the percentage with high counts of Streptococcus mutans was 70% and low counts 30% (chi2 = 26.16 ; p < 0.01). At the laser power of 4,5 watts, the zirconium tip (Z6) had a higher number of sterile culture (96,3%). The numbers of sterile cultures were higher at the laser power of 5 watts, with both tips: zirconium and sapphire, the percentage being equal: 91-92%.