Assessment of multispecies biofilm growth on root canal dentin under different radiation therapy regimens.

OBJECTIVES: To assess the growth of a multispecies biofilm on root canal dentin under different radiotherapy regimens. MATERIALS AND METHODS: Sixty-three human root dentin cylinders were distributed into six groups. In three groups, no biofilm was formed (n = 3): NoRT) non-irradiated dentin; RT55) 55 Gy; and RT70) 70 Gy. In the other three groups (n = 18), a 21-day multispecies biofilm (Enterococcus faecalis, Streptococcus mutans, and Candida albicans) was formed in the canal: NoRT + Bio) non-irradiated + biofilm; RT55 + Bio) 55 Gy + biofilm; and RT70 + Bio) 70 Gy + biofilm. The biofilm was quantified (CFUs/mL). Biofilm microstructure was assessed under SEM. Microbial penetration into dentinal tubules was assessed under CLSM. For the biofilm biomass and dentin microhardness pre- and after biofilm growth assessments, 45 bovine dentin specimens were distributed into three groups (n = 15): NoRT) non-irradiated + biofilm; RT55 + Bio) 55 Gy + biofilm; and RT70 + Bio) 70 Gy + biofilm. RESULTS: Irradiated specimens (70 Gy) had higher quantity of microorganisms than non-irradiated (p = .010). There was gradual increase in biofilm biomass from non-irradiated to 55 Gy and 70 Gy (p < .001). Irradiated specimens had greater reduction in microhardness after biofilm growth. Irradiated dentin led to the growth of a more complex and irregular biofilm. There was microbial penetration into the dentinal tubules, regardless of the radiation regimen. CONCLUSION: Radiotherapy increased the number of microorganisms and biofilm biomass and reduced dentin microhardness. Microbial penetration into dentinal tubules was noticeable. CLINICAL RELEVANCE: Cumulative and potentially irreversible side effects of radiotherapy affect biofilm growth on root dentin. These changes could compromise the success of endodontic treatment in oncological patients undergoing head and neck radiotherapy.

Evaluation of the Antimicrobial Effect of Photodynamic Therapy and Er:YAG Laser Irradiation on Root Canals Infected with Enterococcus faecalis.

Objective: To carry out a histological and morphometric analysis of the antimicrobial effect of Er:YAG laser irradiation combined with photodynamic therapy (PDT) on root canals infected with Enterococcus faecalis. Background: PDT and Er:YAG laser irradiation may be alternatives for effective endodontic disinfection but there are no data on the combination of these therapies. Materials and methods: Forty single-rooted bovine teeth had their roots contaminated with E. faecalis for 72 h. The teeth were randomly divided into four groups (n = 10): group 1, irrigation with 2.5% sodium hypochlorite (NaOCl); group 2, Er:YAG laser (λ2940 nm, 15 Hz, 100 mJ); group 3, PDT with 0.07% methylene blue as photosensitizer and laser irradiation (λ660 nm, power 40 mW, 5 min); and group 4, Er:YAG laser + PDT. After treatment, the teeth were examined by confocal laser scanning microscopy to verify bacterial viability, and morphometric analysis of the images was performed. Results: The PDT and Er:YAG + PDT treatments promoted the greatest reduction in bacteria among the proposed therapies, whereas 2.5% NaOCl was the least effective in bacterial elimination. A statistically significant difference (p < 0.05) was observed among the groups studied, except between the group combining Er:YAG and PDT and the group treated with PDT alone. Conclusions: PDT combined or not with Er:YAG laser was found to be more effective in root canal disinfection when compared with the other groups.

Antimicrobial Photodynamic therapy enhanced by the peptide aurein 1.2.

In the past few years, the World Health Organization has been warning that the post-antibiotic era is an increasingly real threat. The rising and disseminated resistance to antibiotics made mandatory the search for new drugs and/or alternative therapies that are able to eliminate resistant microorganisms and impair the development of new forms of resistance. In this context, antimicrobial photodynamic therapy (aPDT) and helical cationic antimicrobial peptides (AMP) are highlighted for the treatment of localized infections. This study aimed to combine the AMP aurein 1.2 to aPDT using Enterococcus faecalis as a model strain. Our results demonstrate that the combination of aPDT with aurein 1.2 proved to be a feasible alternative capable of completely eliminating E. faecalis employing low concentrations of both PS and AMP, in comparison with the individual therapies. Aurein 1.2 is capable of enhancing the aPDT activity whenever mediated by methylene blue or chlorin-e6, but not by curcumin, revealing a PS-dependent mechanism. The combined treatment was also effective against different strains; noteworthy, it completely eliminated a vancomycin-resistant strain of Enterococcus faecium. Our results suggest that this combined protocol must be exploited for clinical applications in localized infections as an alternative to antibiotics.

Susceptibility of Enterococcus faecalis and Propionibacterium acnes to antimicrobial photodynamic therapy.

Bacterial resistance to available antibiotics nowadays is a global threat leading researchers around the world to study new treatment modalities for infections. Antimicrobial photodynamic therapy (aPDT) has been considered an effective and promising therapeutic alternative in this scenario. Briefly, this therapy is based on the activation of a non-toxic photosensitizing agent, known as photosensitizer (PS), by light at a specific wavelength generating cytotoxic singlet oxygen and free radicals. Virtually all studies related to aPDT involve a huge screening to identify ideal PS concentration and light dose combinations, a laborious and time-consuming process that is hardly disclosed in the literature. Herein, we describe an antimicrobial Photodynamic Therapy (aPDT) study against Enterococcus faecalis and Propionibacterium acnes employing methylene blue, chlorin-e6 or curcumin as PS. Similarities and discrepancies between the two bacterial species were pointed out in an attempt to speed up and facilitate futures studies against those clinical relevant strains. Susceptibility tests were performed by the broth microdilution method. Our results demonstrate that aPDT mediated by the three above-mentioned PS was effective in eliminating both gram-positive bacteria, although P. acnes showed remarkably higher susceptibility to aPDT when compared to E. faecalis. PS uptake assays revealed that P. acnes is 80 times more efficient than E. faecalis in internalizing all three PS molecules. Our results evidence that the cell wall structure is not a limiting feature when predicting bacterial susceptibility to aPDT treatment.

Effect of different diode laser wavelengths on root dentin decontamination infected with Enterococcus faecalis.

The objective of this study was to evaluate the antibacterial effect and the ultrastructural alterations of diode laser with different wavelengths (808nm and 970nm) and its association with irrigating solutions (2.5% sodium hypochlorite and 2% chlorhexidine) in root dentin contaminated by a five days biofilm. Thirteen uniradicular teeth were sectioned into 100 dentin intraradicular blocks. Initially, the blocks were immersed for 5min in 17% EDTA and washed with distilled water for 5min, then samples were sterilized for 30min at 120°C. The dentin samples were inoculated with 0.1mL of E. faecalis suspension in 5mL BHI (Brain Heart Infusion) and incubated at 37°C for 5days. After contamination, the specimens were distributed into ten groups (n=10) according to surface treatment: GI - 5mL NaOCl 2.5%, GII - 5mL NaOCl 2.5%+808nm diode (0.1W for 20s), GIII - 5mL NaOCl 2.5%+970nm diode (0.5W for 4s), GIV - 808nm diode (0.1W for 20s), GV - 970nm diode (0.5W for 4s), GVI - CHX 2%, GVII - CHX 2%+808nm diode (0.1W for 20s), GVIII - CHX 2%+970nm diode (0.5W for 4s), GIX - positive control and GX - negative control. Bacterial growth was analyzed by turbidity and optical density of the growth medium by spectrophotometry (nm). Then, the specimens were processed for analysis ultrastructural changes of the dentin surface by SEM. The data was subject to the One-way ANOVA test. GI (77.5±12.1), GII (72.5±12.2), GIII (68.7±8.7), GV (68.3±8.7), GVI (62.0±5.5) and GVII (67.5±3.3) were statistically similar and statistically different from GIV (58.8±25.0), GVIII (59.2±4.0) and control groups (p<0.05). SEM analysis showed a modified amorphous organic matrix layer with melted intertubular dentin when dentin samples were irradiated with 970nm diode laser; erosion of the intertubular dentin in blocks submitted to 808nm diode laser irradiation; and an increased erosion of the intertubular dentin when 2.5% NaOCl was associated to the different wavelengths lasers. All the therapeutic protocols were able to reduce the bacterial contingent in dentin blocks, and the association of diode laser and solutions did not significantly improve the reduction of the bacterial contingent.

Influence of sucrose on growth and sensitivity of Candida albicans alone and in combination with Enterococcus faecalis and Streptococcus mutans to photodynamic therapy.

This study has evaluated the effects of photodynamic inactivation (PDI) using erythrosine as photosensitizer and green light-emitting diode (LED) on biofilms of Candida albicans alone and in combination with Enterococcus faecalis and Streptococcus mutans. We have also evaluated the effect of sucrose on biofilm formation and bacterial growth and sensitivity to PDI. Biofilms were formed in suspension of 106 cells/ml on plates before being grown in broth culture with and without sucrose and incubated for 48 h. Next, the treatment was applied using erythrosine at a concentration of 400 µM for 5 min and green LED (532 ± 10 nm) for 3 min on biofilms alone and in combination. The plates were washed and sonicated to disperse the biofilms, and serial dilutions were carried and aliquots seeded in Sabouraud agar before incubation for 48 h. Next, the colony-forming units per milliliter (CFU/ml; log10) were counted and analyzed statistically (ANOVA, Tukey test, P ≤ 0.05). Results show that S. mutans favors the growth of C. albicans in biofilms with sucrose, with treatment not being effective. However, when the biofilm was grown without sucrose, we found a reduction in biofilm formation and a significant decrease in the PDI treatment (P < 0.0001). In conclusion, both growth and sensitivity to PDI in biofilms of C. albicans are strongly influenced by bacterial combination, and the presence of sucrose affected directly the growth and sensitivity of the biofilm to PDI as sucrose is the substrate for construction of the exopolysaccharide matrix.

Antimicrobial Activity of Photodynamic Therapy Against Enterococcus faecalis Before and After Reciprocating Instrumentation in Permanent Molars.

OBJECTIVE: The present study sought to evaluate the antimicrobial activity against Enterococcus faecalis of photodynamic therapy applied before and after reciprocating instrumentation of permanent molars. BACKGROUND: Apical extrusion of debris can cause flare-ups due to introduction of bacteria into the periapical tissues. METHODS: Eighteen mesial roots from permanent mandibular molars were selected. The crowns were removed to obtain a standard root length of 15 mm. The included mesial roots had an angulation of 10°-40° and canals with independent foramina. The orifice of each mesiolingual canal was sealed with light-curing resin, and the working length was established visually, 1 mm short of the apical foramen. The roots were rendered impermeable and sterilized, and the mesiobuccal canals were contaminated with a standard strain of E. faecalis for 21 days. Specimens were randomly divided into three groups (n = 6): G1, photodynamic therapy performed before instrumentation and irrigation with 0.9% NaCl (saline) solution; G2, photodynamic therapy performed after instrumentation and irrigation with 0.9% NaCl; and G3 (control), instrumentation and irrigation with 2.5% NaOCl (sodium hypochlorite) solution. Canals were shaped with a WaveOne primary file (25.08) and irrigated with 0.9% NaCl. E. faecalis samples were collected before and after each procedure, and the results were analyzed using descriptive statistics and the Kruskal-Wallis and Wilcoxon tests. RESULTS: Significant reductions in E. faecalis were observed when photodynamic therapy was performed before and after instrumentation of the root canal system (p < 0.05). Reciprocating instrumentation significantly reduced E. faecalis colonies in experimentally contaminated root canal systems (p < 0.05). CONCLUSIONS: Photodynamic therapy was effective in removing E. faecalis from the root canal system, whether performed before or after reciprocating instrumentation.

Ação antimicrobiana da terapia fotodinâmica sobre Enterococcus faecalis realizada pela combinação do aparelho fotopolimerizador a LED e curcumina / Antimicrobial activity of the combination of LED curing light and curcumin on Enterococcus faecalis in photodynamic therapy

Este trabalho avaliou a ação antimicrobiana da terapia fotodinâmica (TFD) realizada pela combinação do aparelho fotopolimerizador (LED 430-480nm) e curcumina 20µM, além de sua associação com Gel Carbopol 2%. Fotos padronizadas, na ausência e presença do gel Carbopol no interior do conduto, previamente preparado, foram analisadas no software OriginLab para verificar a manutenção da intensidade de luz ao longo do canal. Na análise antimicrobiana, uma linhagem de ATCC 4083 do Enterococcus faecalis foi utilizada para formação de biofilme sobre a superfície de discos de dentina (4mm de diâmetro) e para contaminação intratubular de segmentos radiculares (15mm), ambos obtidos de raízes bovinas. A ação sobre o biofilme foi avaliada de acordo com os grupos (n=20): [BST]: sem tratamento (controle); [BC]: Curcumina; [BL]: LED; [BCL]: Curcumina + LED e [BAL]: Azul de metileno 0.05% + LASER (660nm). A ação intratubular foi avaliada nos grupos (n=8): [IST]: sem tratamento (controle); [ICL]: Curcumina + LED; [ICGL]: Curcumina + Gel Carbopol + LED e [IAL]: Azul de metileno 0.05% + LASER (660nm). Discos e segmentos foram analisados em Microscópio Confocal de Varredura a LASER para detectar a porcentagem de bactérias viáveis. Os dados obtidos, que apresentaram distribuição normal, foram analisados estatisticamente pelo teste ANOVA, a 1 critério, e para distribuição não normal, Kruskall- Wallis. Os resultados mostraram que a presença do gel Carbopol no conduto não contribuiu na manutenção da intensidade da luz LED. Sobre o biofilme, os grupos BAL e BCL apresentaram maior redução microbiana, sem diferença entre eles, enquanto que, na ação intratubular, o IAL apresentou a melhor ação antimicrobiana, com diferença estatística com os demais. Concluiuse que a TFD realizada com o fotopolimerizador e curcumina é tão eficaz quanto a realizada com LASER e azul de metileno. Entretanto, é necessário investigar meios que permitam a sua utilização no interior do conduto radicular, já que o Gel Carbopol não manteve a intensidade da luz ao longo do trajeto.(AU)

This work aimed to evaluate the antimicrobial action of photodynamic therapy (PDT) by the combination of light curing device (LED 430-480nm) and Curcumin 20 µM, and its in association with Carbopol Gel 2%. Standard photos in the absence and presence of Carbopol gel inside of the root canal previously prepared, were analyzed in OriginLab software to check the light intensity maintenance along the canal. In microbial analysis, a strain of Enterococcus faecalis ATCC 4083 was used to make biofilm on the surface of dentin disks (4 mm in diameter), and intratubular contamination of root segments (15 mm), both obtained from bovine roots. The action on the biofilm was evaluated according to groups (n = 20): [BNT]: no treatment (control); [BC]: Curcumin; [BL]: LED; [BCL]: Curcumin + LED and [BML]: methylene blue 0.05% + LASER (660nm). Intratubular action was evaluated in groups (n = 8): [INT]: no treatment (control); [ICL]: Curcumin + LED; [ICGL]: Curcumin Carbopol Gel + LED + and [IML]: methylene blue 0.05% + LASER (660nm). Disks and segments were analyzed in LASER Scanning Confocal Microscope to detect the percentage of viable bacteria. The data, which showed normal distribution, were statistically analyzed by ANOVA 1 WAY test, and for no normal distribution, Kruskall-Wallis. The results showed that presence of Carbopol gel in the root canal did not increase the maintenance of intensity of LED light. About the biofilm, BML and BCL had higher microbial reduction, no difference between them, while in action intratubular, IML presented the higher antimicrobial action, with statistical difference with the other. It was concluded that the PDT realized with curing light and Curcumin is as effective as LASER and methylene blue. However, it is necessary to investigate ways that allow its use inside the root canal, because the Carbopol gel did not maintain the intensity of light until apex.(AU)

Biofilm formation on polystyrene under different temperatures by antibiotic resistant Enterococcus faecalis and Enterococcus faecium isolated from food.

The ability of antibiotic resistant E. faecalis and E. faecium isolated from food to form biofilm at different temperatures in the absence or presence of 0.75% glucose was evaluated. A synergistic effect on biofilm at 10 °C, 28 °C, 37 °C and 45 °C and glucose was observed for E. faecalis and E. faecium.

The use of optical fiber in endodontic photodynamic therapy. Is it really relevant?

This study analyzed the necessity of use of an optical fiber/diffusor when performing antimicrobial photodynamic therapy (PDT) associated with endodontic therapy. Fifty freshly extracted human single-rooted teeth were used. Conventional endodontic treatment was performed using a sequence of ProTaper (Dentsply Maillefer Instruments), the teeth were sterilized, and the canals were contaminated with Enterococcus faecalis 3 days' biofilm. The samples were divided into five groups: group 1--ten roots irradiated with a laser tip (area of 0.04 cm(2)), group 2--ten roots irradiated with a smaller laser tip (area of 0.028 cm(2)), and group 3--ten teeth with the crown, irradiate with the laser tip with 0.04 cm(2) of area. The forth group (G4) followed the same methodology as group 3, but the irradiation was performed with smaller tip (area of 0.028 cm(2)) and G5 ten teeth with crown were irradiated using a 200-mm-diameter fiber/diffusor coupled to diode laser. Microbiological samples were taken after accessing the canal, after endodontic therapy, and after PDT. Groups 1 and 2 showed a reduction of two logs (99%), groups 3 and 4 of one log (85% and 97%, respectively), and group 5 of four logs (99.99%). Results suggest that the use of PDT added to endodontic treatment in roots canals infected with E. faecalis with the optical fiber/diffusor is better than when the laser light is used directed at the access of cavity.

Ação da clorofila como fotossensibilizador e do LED como fonte de luz alternativa na terapia fotodinâmica antimicrobiana contra o Enterococcusfaecalis / Effect of chlorophyll as photosensitizer and LED as an alternative light source on Antimicrobial Photodynamic Therapy against Enterococcus faecalis

O objetivo deste estudo foi investigar o efeito da Terapia Fotodinâmica antimicrobiana (TFDa) sobre o Enterococcus faecalis (E. faecalis), in vitro, utilizando a clorofila (CL) como agente fotossensibilizador (FS) e o diodo emissor de luz (LED) como fonte de luz. A cultura pura de E. faecalis foi ativada em caldo de BHI a 37oC por 24h. O cultivo do microrganismo foi centrifugado a 3000rpm por 15min, e o pellet re-suspenso em 0,85% de solução salina. As concentrações da bactéria foram ajustadas para 107UFC mL-1 (unidades formadoras de colônias por mililitro). Diferentes tipos de solventes para a CL foram testados: éter, álcool de cereais, Tween 80 e P-123. 100l do inóculo e o mesmo volume da solução teste foram inseridos em cada poço da placa de microtitulação. A mistura foi agitada e aguardou-se 1min. 25l da suspensão foi removida e diluições seriadas foram realizadas. Alíquotas de cada diluição foram espalhadas na superfície da placa, armazenadas em microaerofilia e incubadas na estufa por 24h a 37oC. O número de UFC foi contado em cada placa. Para o experimento da TFDa, os grupos foram divididos em: controle (G1); TBO 1min (G2); CL+Tween 1min (G3); CL+Tween 5min (G4); CL+P-123 1min (G5); CL+P-123 5min (G6); CL+Tween 1min + LED 1min (G7); CL+Tween 1min + LED 5min (G8); CL+Tween 5min + LED 1min (G9); CL+Tween 5min + LED 5min (G10); CL+P-123 1min + LED 1min (G11); CL+P-123 1min + LED 5min (G12); CL+P-123 5min + LED 1min (G13); CL+P-123 5min + LED 5min (G14); TBO 1min + LED 1min (G15); TBO 1min + LED 5min (G16); LED 1min (G17); LED 5min (G18).Um volume de 100l da suspensão bacteriana foi inserido em poços da placa e o mesmo volume da solução do FS foi adicionado. A CL foi solubilizada em soluções aquosas de Tween ou P-123. Cada poço foi agitado e o tempo de préirradiação foi de 1 ou 5min. O LED foi acionado durante 1 ou 5min. 25l da suspensão foi submetida a diluições seriadas e as alíquotas de cada diluição foram espalhadas na superfície da placa em Ágar BHI...

The aim of this study was to investigate the effect of Antimicrobial Photodynamic Therapy (aPDT) against Enterococcus faecalis (E. faecalis), in vitro, using chlorophyll (CL) as photosensitizer (FS) agent and light-emitting diode (LED) as light source. Pure culture of E. faecalis was cultivated in BHI broth at 37oC for 24h. The culture was centrifuged at 3000rpm for 15min, and the pellets were resuspended in 0.85% saline solution. The bacterial concentrations were adjusted to 107CFU mL-1 (colony forming units per milliliter). Different types of solvents for CL were tested: ether, grain alcohol, Tween 80 and P-123. 100l of inoculum and the same volume of the test solution were inserted into each well of the microtitre plate. The mixture was mixed and 1 min was awaited. 25l of the suspension was removed and serial dilutions were performed. Aliquots of each dilution were spread on the surface of the plates, stored in microaerophilic conditions and incubated for 24h at 37oC. The number CFU was counted in each plate. For aPDT experiment, the groups were divided into: control (G1); TBO 1min (G2); CL+Tween 1min (G3); CL+Tween 5min (G4); CL+P-123 1min (G5); CL+P-123 5min (G6); CL+Tween 1min + LED 1min (G7); CL+Tween 1min + LED 5min (G8); CL+Tween 5min + LED 1min (G9); CL+Tween 5min + LED 5min (G10); CL+P-123 1min + LED 1min (G11); CL+P-123 1min + LED 5min (G12); CL+P-123 5min + LED 1min (G13); CL+P-123 5min + LED 5min (G14); TBO 1min + LED 1min (G15); TBO 1min + LED 5min (G16); LED 1min (G17); LED 5min (G18). A volume of 100l of bacterial suspension was inserted into the well of microtitre plate and the same volume of FS was added. CL was solubilized in aqueous solution of Tween and P-123. Each well was mixed and preirradiation time was 1 or 5min. LED was irradiated during 1 or 5 min. 25l of suspension was subjected to serial dilutions and aliquots of each dilution were spread on the surface of agar BHI plates. The plates were stored and incubated at 37oC for 24h...

Biofilm formation on polystyrene under different temperatures by antibiotic resistant Enterococcus faecalis and Enterococcus faecium isolated from food

The ability of antibiotic resistant E. faecalis and E. faecium isolated from food to form biofilm at different temperatures in the absence or presence of 0.75% glucose was evaluated. A synergistic effect on biofilm at 10 °C, 28 °C, 37 °C and 45 °C and glucose was observed for E. faecalis and E. faecium.

Bactericidal effects of two parameters of Er:YAG laser intracanal irradiation: ex-vivo study.

The success of endodontic treatment depends on the complete elimination of microorganisms from the root canal system, thus the search for new procedures to eliminate them is justified. The aim of this study was to assess bacterial reduction after intracanal irradiation with the Er:YAG laser. The canals of 70 extracted human maxillary canines were prepared up to file #40 using 1% NaOCl, irrigated with 17% EDTA, and then washed with physiological solution activated by ultrasound. The roots were sterilized by autoclaving, inoculated with 10 µl of a suspension containing 1.5 × 10(8) CFU/ml of Enterococcus faecalis ATCC 29212 and incubated at 37°C for 72 h. The canals were irradiated with the Er:YAG laser using two energy settings: 60 mJ and 15 Hz, and 100 mJ and 10 Hz. The remaining bacteria were counted immediately and 48 h after laser irradiation. The results showed a high bacterial reduction at both time points. With 60 mJ and 15 Hz there was an immediate reduction of 99.73% and the reduction was 77.02% after 48 h, and with 100 mJ and 10 Hz there was an immediate reduction of 99.95% and the reduction was 84.52% after 48 h. Although the best results were observed with 100 mJ of energy, the difference between the two settings was not statistically significant. The count performed 48 h after irradiation showed that E. faecalis were able to survive, and can grow even from small numbers.

Photodynamic therapy with two different photosensitizers as a supplement to instrumentation/irrigation procedures in promoting intracanal reduction of Enterococcus faecalis.

INTRODUCTION: This in vitro study aimed to investigate the antibacterial effects of photodynamic therapy (PDT) with methylene blue (MB) or toluidine blue (TB) (both at 15 microg/mL) as a supplement to instrumentation/irrigation of root canals experimentally contaminated with Enterococcus faecalis. METHODS: Seventy extracted teeth had their root canals contaminated with an endodontic strain of E. faecalis for 7 days, instrumented with nickel-titanium instruments and irrigated either with 2.5% NaOCl or with 0.85% NaCl, and then randomly distributed into four experimental groups: MB/NaOCl (PDT with MB and NaOCl as the irrigant), TB/NaOCl (PDT with TB and NaOCl as the irrigant), MB/NaCl (PDT with MB and NaCl as the irrigant), and TB/NaCl (PDT with TB and NaCl as the irrigant). For PDT, the photosensitizer remained in the canal for 2 minutes before exposed to red light emitted from a diode laser for 4 minutes. Samples were taken before and after instrumentation/irrigation and following the specific PDT procedure for each group, plated onto Mitis-salivarius agar and the colony forming units counted. RESULTS: Regardless of the irrigant used (NaOCl or NaCl), instrumentation significantly reduced bacterial counts in comparison to the baseline (p < 0.001). NaOCl as the irrigant was significantly more effective than NaCl, and this difference persisted after PDT, irrespective of the photosensitizer used (p < 0.05). PDT with either MB or TB did not significantly enhance disinfection after chemomechanical preparation using NaOCl as irrigant (p > 0.05). No significant differences were observed between the two photosensitizers (p > 0.05). CONCLUSION: These in vitro results suggest that PDT with either MB or TB may not exert a significant supplemental effect to instrumentation/irrigation procedures with regard to intracanal disinfection. Further adjustments in the PDT protocol may be required to enhance predictability in bacterial elimination before clinical use is recommended.

Effectiveness of 980-mm diode and 1064-nm extra-long-pulse neodymium-doped yttrium aluminum garnet lasers in implant disinfection.

OBJECTIVE: To evaluate the potential of 980-nm gallium aluminum arsenide (GaAlAs) and 1064-nm neodymium-doped yttrium aluminum garnet (Nd:YAG) lasers to reduce bacteria after irradiation of implant surfaces contaminated with Enterococcus faecalis and Porphyromonas gingivalis and on irradiated implant surface morphology. BACKGROUND: Despite the frequency of implant success, some implant loss is related to peri-implantitis because of difficulty in eliminating the biofilm. METHODS: Implants (3.75 x 13 mm) with machined surfaces, surfaces sand blasted with titanium oxide (TiO(2)), and sand-blasted and acid-etched surfaces were exposed to P. gingivalis and E. faecalis cultures and irradiated with 980-nm GaAlAs or 1064-nm Nd:YAG lasers. After laser treatments, the number of remaining colony-forming units and implant surface morphology were analyzed using scanning electron microscopy (SEM). RESULTS: The Nd:YAG laser was able to promote a total contamination reduction on all implants irradiated. The results with the GaAlAs laser showed 100% bacteria reduction on the implants irradiated with 3 W. Irradiation with 2.5 W and 3 W achieved 100% of bacteria reduction on P. gingivalis-contaminated implants. Decontamination was not complete for the sand-blasted TiO(2) (78.6%) and acid-etched surfaces (49.4%) contaminated with E. faecalis and irradiated with 2.5 W. SEM showed no implant surface changes. CONCLUSION: The wavelengths used in this research provided bacteria reduction without damaging implant surfaces. New clinical research should be encouraged for the use of this technology in the treatment of peri-implantitis.

Utilização da terapia fotodinâmica associada ou não ao hipoclorito de sódio na instrumentação de canais radiculares contaminados com Enterococcus faecalis / The use of photodynamic therapy associated or not to sodium hypochlorite in the instrumentation of radicular canals contaminated by Enterococcus faecalis

O presente trabalho teve por objetivo avaliar a ação do laser de baixa intensidade associado ao corante azul de toluidina, como agente bactericida durante a fase de instrumentação dos canais radiculares, comparando sua utilização associada ou não ao hipoclorito de sódio 0,5%. Para tal, 90 caninos humanos foram instrumentados, autoclavados e 88 imersos em caldo TSB (Tryptcase Soy Broth). Em seguida foram inoculados com suspensão de E. faecalis e incubados em estufa a 37°C, por 72 horas, para permitir a formação do biofilme. As amostras foram randomicamente divididas em 4 grupos de 22 dentes cada e 1 grupo controle negativo com 2 dentes. Grupo I: reinstrumentados e irrigados com soro fisiológico com posterior aplicação de terapia fotodinâmica (PDT); Grupo II: reinstrumentados e irrigados com hipoclorito de sódio a 0,5% com posterior aplicação de PDT; Grupo III: reinstrumentados e irrigados com hipoclorito de sódio a 0,5%; Grupo IV controle positivo: não foi realizado nenhum tipo de tratamento antes da coleta do material e Grupo V controle negativo: não foram contaminados. Para a coleta da dentina intracanal utilizou-se uma broca Gates Glidden número 6. A atividade antimicrobiana foi avaliada por meio da contagem de UFCs (unidades formadoras de colônias). Os dentes foram então imersos em meio seletivo para enterococos, incubados em estufa a 37º C por 72 horas e avaliados quanto à alteração de coloração do meio. Todas as amostras, exceto o controle negativo, apresentaram-se positivas. Com a finalidade de certificar-se da formação do biofilme e da confirmação dos resultados das contagens das UFCs, 9 amostras foram avaliadas ao MEV (microscopia eletrônica de varredura)...

This current paper aims to assess the action of low-intensity laser together with toluidine blue as a bactericidal agent during the instrumentation phase of radicular canals, comparing its associated or non-associated use with sodium hypochlorite 0.5%. To do so, 90 human canines were instrumented, autoclaved, and 88 were immersed into TSB, and were then inoculated with an E. faecalis suspension and brought into an incubator at 37 degrees Celsius for 72 hours in order to allow the formation of biofilm. The samples were randomly divided into 5 groups, 4 of which composed of 22 samples each plus 1 negative control. Group I: reinstrumented and irrigated with saline solution and subsequent application of photodynamic therapy (PDT); Group II: reinstrumented and irrigated with sodium hypochlorite 0.5% and subsequent PDT application; Group III: reinstrumented and irrigated with sodium hypochlorite 0.5%; Group IV: positive control (no previous treatment before material collection); Group V: negative control (2 samples) with no contamination. The collection of intracanal dentin was obtained by using a Gates Glidden drill nr. 6. The antimicrobial activity was assessed by CFUs counting. The teeth were then immersed in a selected medium for Enterococcus, incubated at 37 degrees Celsius for 72 hours. They were then assessed for medium turbidity. All samples, with the exception of the negative control, showed positive. In order to make sure of the formation of biofilm and the confirmation of the CFUs counting, 9 samples were scavenged under the electron microscope...

High-power diode laser in the disinfection in depth of the root canal dentin.

OBJECTIVE: The objective of this study was to evaluate the disinfection degree of dentine caused by the use of diode laser after biomechanical procedures. STUDY DESIGN: Thirty teeth were sectioned and roots were autoclaved and incubated for 4 weeks with a suspension of Enterococcus faecalis. The specimens were randomly divided into 3 groups (n = 10): G1, instrumented with rotary files, irrigated with 0.5% sodium hypochlorite and 17% EDTA-T, and then irradiated by 830-nm diode laser at 3 W; G2, the same procedures as G1 but without laser irradiation; and G3, irrigation with saline solution (control). Dentin samples of each third were collected with carbide burs and aliquots were sowed to count viable cells. RESULTS: The disinfection degree achieved was 100% in G1 and 98.39% in G2, when compared to the control group (G3). CONCLUSION: Diode laser irradiation provided increased disinfection of the deep radicular dentin in the parameters and samples tested.

Disinfection of root canals using Er:YAG laser at different frequencies.

OBJECTIVE: This study evaluated, in vitro, the degree of disinfection of the Er:YAG laser in root canals contaminated with Bacillus subtilis, Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, and Candida albicans, for 28 days. METHODS: Forty-six single-rooted human teeth were divided into five groups of eight teeth each; three teeth were used as negative controls and three as positive controls. After contamination, the root canals were prepared mechanically. Three groups were irradiated with Er:YAG laser at 100 mJ, varying the frequency (7, 10, and 16 Hz). Two groups were irrigated with 1.0% and 2.5% NaOCl solution. After treatment, two sterilized paper cones were placed in the root canals for 5 min. One cone was transferred to 2.0 mL of Letheen broth culture medium, incubated at 37 degrees C for 48 h, and then 0.1 mL of that solution was placed in 2.0 mL of brain heart infusion for 48 h to determine microbial growth. The other cone was transferred to a test pipette with peptone and water for serial dilution and spread in Müeller Hinton medium. After 24 h of incubation, the colony-forming units (CFUs) were counted. RESULTS: There was a microbial reduction of 85.33% for the group irradiated with Er:YAG laser at 100 mJ/7 Hz, 74.58% at 100 mJ/10 Hz, and 89.50% at 100 mJ/16 Hz. For the groups irrigated with 1.0% and 2.5% NaOCl solution, 83.15% and 84.46% values of microbial reduction were obtained respectively. CONCLUSION: All the groups showed statistically similar results (p > 0.05%). No method totally eliminated microorganisms.

Evaluation of the antimicrobial effect of Er:YAG laser irradiation versus 1% sodium hypochlorite irrigation for root canal disinfection.

This laboratory study evaluated Er:YAG laser antibacterial action in infected root canals. Forty-eight maxillary central incisors were used. After canal preparation, the teeth were autoclaved and divided into four groups: (1) non-treated teeth (control group); (2) teeth treated with NaOCl; (3) teeth irradiated with Er:YAG laser (7 Hz, 100 mJ, 80 pulses/canal, 11 sec) to the working length; (4) teeth irradiated similarly to, but 3 mm short, of the apex. The root canals from Groups 2, 3 and 4 were inoculated with 4 bacteria: Bacillus subtillus, Enterococcus faecalis, Pseudomonas aeruginosa, and Staphylococcus aureus, together with Candida albicans, and maintained for 24 h at 37 degrees C. All suspensions were adjusted to tube 2 of the MacFarland scale. The intracanal material was then collected with sterile paper points, which were placed in the canals for 5 min and then immersed in 5 ml of BHI medium. This was then seeded onto agar and stained by Gram's method. The NaOCl solutions and the Er:YAG laser irradiation to working length were effective against all five micro-organisms; however, 70% of the specimens irradiated 3 mm short of the apex remained infected.

The bactericidal effect of Ho:YAG laser irradiation within contaminated root dentinal samples.

OBJECTIVE: This in vitro study investigates the bactericidal effect of pulsed Ho:YAG laser irradiation in the depth of contaminated dentin specimens. BACKGROUND DATA: Previous studies have shown the effectiveness of laser irradiation in bacterial reduction of infected root canal. METHODS: Root dentin of bovine teeth were sliced longitudinally in 180 samples of 100 microm, 300 microm, and 500 microm thickness, sterilized, dried, and inoculated on one side, with 1 microL of Enterococcus faecalis suspension. The opposite side's were irradiated four times for 5 seconds each with Ho:YAG laser irradiation, a wavelength of 2.10 microm, using four different energy settings: 1 W/5 Hz; 1 W/10 Hz; 1.5 W/5 Hz, and 2.0 W/5 Hz through a 320-microm quartz fiber at an angle of approximately 5 degrees. In addition, two control groups were investigated, the first was inoculated and not submitted to any treatment, the second was inoculated and treated with NaOCl and H2O2. The remaining bacteria from each dentin sample in a transport media were removed by vibration, serially diluted, and plated out on culture dishes selective for Enterococcus faecalis. RESULTS: When compared with the untreated control group or even with the group treated with NaOCl plus H2O2, counting of colonies forming units (CFU) from the laser-treated samples revealed a high significant bacterial elimination with a maximum of 98.46% and a minimum of 83.65%. CONCLUSIONS: Our findings demonstrate a significant decrease of the bacterial population in depth, suggesting that the Ho:YAG laser irradiation could be effective to eliminate the microorganisms harbored within dentin or contaminated canals.