Studying the viability and growth kinetics of vancomycin-resistant Enterococcus faecalis V583 following femtosecond laser irradiation (420-465 nm).

Enterococcus faecalis is among the most resistant bacteria found in infected root canals. The demand for cutting-edge disinfection methods has rekindled research on photoinactivation with visible light. This study investigated the bactericidal activity of femtosecond laser irradiation against vancomycin-resistant Enterococcus faecalis V583 (VRE). The effect of parameters such as wavelength and energy density on the viability and growth kinetics of VRE was studied to design an optimized laser-based antimicrobial photoinactivation approach without any prior addition of exogenous photosensitizers. The most effective wavelengths were 430 nm and 435 nm at a fluence of 1000 J/cm2, causing a nearly 2-log reduction (98.6% and 98.3% inhibition, respectively) in viable bacterial counts. The colony-forming units and growth rate of the laser-treated cultures were progressively decreased as energy density or light dose increased at 445 nm but reached a limit at 1250 J/cm2. At a higher fluence of 2000 J/cm2, the efficacy was reduced due to a photobleaching phenomenon. Our results highlight the importance of optimizing laser exposure parameters, such as wavelength and fluence, in bacterial photoinactivation experiments. To our knowledge, this is the first study to report an optimized wavelength for the inactivation of VRE using visible femtosecond laser light.

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.

Photodynamic Therapy With Resveratrol and an Nd:YAG Laser for Enterococcus faecalis Elimination.

BACKGROUND/AIM: Enterococcus faecalis is the leading cause of endodontic treatment failures. Despite various conventional disinfection approaches, microorganisms often persist in root canals. Photodynamic therapy (PDT) is an adjunct antimicrobial strategy employing a nontoxic photosensitizer (PS) and light source. This study evaluated the antimicrobial effect of PDT using an Nd:YAG laser and resveratrol (RSV) with or without pigment, and confirmed that RSV is nontoxic as a PS. MATERIALS AND METHODS: We employed laser irradiation at a 3W output power, using RSV and red pigment as the PS, on an E. faecalis bacterial solution. Subsequently, colony-forming units were quantified. The impact of RSV on osteoblasts was measured using an MTT assay. RESULTS: E. faecalis counts declined after laser irradiation. The combined application of laser irradiation with RSV, red pigment, or both showed a reduction compared to no irradiation and control groups without RSV and red pigment. The 50% cytotoxic concentration against osteoblast cells from mice incubated with RSV for 48 h was 162 µM. The value with RSV and laser was 201 µM and that with RSV and red pigment was 199 µM. The value with RSV, laser and red pigment was 357 µM. CONCLUSION: The combination of Nd:YAG laser irradiation and RSV as the PS with pigment was efficacious for E. faecalis elimination without inducing any toxic effects on osteoblasts. This combination holds potential as a root canal irrigation strategy.

An in vitro study on the efficacy of hydrogen peroxide mediated high-power photodynamic therapy affecting Enterococcus faecalis biofilm formation and dispersal.

OBJECTIVE: Biofilms are involved in failure of root canal treatment due to their high resistance to antimicrobial agents, which make their removal as a big challenge. The present study aims at utilizing hydrogen peroxide (HP) plus high frequency laser reinforced antimicrobial photodynamic therapy (a-PDT) as a complementary therapy against Enterococcus faecalis (E. faecalis) at planktonic and biofilm stages. MATERIALS AND METHODS: E. faecalis at planktonic and biofilm stages was treated with the photosensitizer HP, followed by no irradiation or irradiation with a power of 2.5 W (ʎ = 980 nm). The cell viability, anti-biofilm, anti-metabolic potential, and temperature changes were evaluated. RESULTS: The combination of HP and 980 nm diode laser intensely boosted antibacterial and anti-biofilm efficacy compared with either component alone, affirming HP reinforcement as a bacteriostatic agent. The maximum effect on biofilm occurs in 5.25% sodium hypochlorite (NaOCl) group. During laser irradiations, the mean of temperature changes remains below 5.6 °C. CONCLUSIONS: It could be concluded that the HP could improve anti-biofilm efficacy as a photosensitizer in a-PDT.

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.

Enhanced disinfection of E. faecalis and levofloxacin antibiotic degradation using tridoped B-Ce-Ag TiO2 photocatalysts synthesized by ecofriendly citrate EDTA complexing method.

Since its use for photochemical water splitting reported first in 1972, TiO2 is one of the most extensively studied photocatalysts for a diverse range of applications. Monodoping or codoping of the catalyst is a proven strategy to enhance the functionality of TiO2 under solar or visible light. However, the use of three or more dopants in the development of more efficient and visible light active photocatalysts has not been investigated widely, especially for microbial disinfection. Boron/cerium/silver tridoped TiO2 photocatalysts with curated amounts of the dopants (B = 1, 2 at.%, Ce = 0.1 at.%, Ag = 0.06 at.%), synthesized by the ecofriendly EDTA-citrate method, were evaluated for the disinfection of water using Enterococcus faecalis under UV-A irradiation and degradation of levofloxacin antibiotic under solar light. The catalyst characterization revealed that the spherical nanoparticles had a crystallite size of ~ 13 nm and bandgap energy values of 2.8-2.9 eV. 2B-0.1Ce-0.06Ag-TiO2 is the best catalyst for microbial disinfection with a log reduction and kinetic rate constant ~ 30 and ~ 4.5 times higher than those values determined for the other codoped or monodoped catalysts, confirming an enhanced performance. Regarding levofloxacin degradation, the best performing catalyst is 1B-0.1Ce-0.06Ag-TiO2 with degradation of 99% and 83% COD reduction in 100 min. The tridoped photocatalysts are very effective in the inactivation of Enterococcus faecalis, thus solving the problem of antimicrobial resistance in waters containing antibiotic residues.

Bactericidal effect of Er,Cr:YSGG laser irradiation on endodontic biofilm: An ex vivo study.

AIM: This ex vivo study aimed to evaluate the of Er,Cr:YSGG laser effectiveness in the decontamination of an endodontic biofilm. MATERIALS AND METHODS: Seventy-three single rooted human teeth, freshly were chosen. Each tooth was exposed to four associated species in an endodontic biofilm (Enterococcus faecalis, Streptococcus salivarius, Porphyromonas gingivalis, and Prevotella intermedia) and randomly allocated to one of the seven experimental groups. The group 1 (7 teeth) was used to finalize the reliable biofilm-forming technique. The groups 2 and 3 (15 teeth each group) were irradiated with two different Er;Cr:YSGG laser settings (0,75 W - 40 Hz and 4 W - 40 Hz, respectively). The groups 4 and 5 (15 teeth each group) were irrigated with two different solutions and laser irradiated with the same settings (1,5 W - 15 Hz). The group 6 (6 teeth) was the control group treated only with 4 ml 2,5% NaOCl irrigation during 60 s. RESULTS: The observations of group 2 and 3 specimens showed the ripeness of the biofilm with the presence of Enterococcus faecalis and Streptococcus salivarius in chains but in group 3 thermal edge effects produced by the optic fiber in the canal walls were present. The group 4 specimens observation showed an average cleaning of the root canal walls while on the canal walls of group 5 samples the apical third presented several debris and smear layer and in the centre cracks and melting dentin of the radicular wall were observed. CONCLUSION: In those experimental conditions, this study, demonstrated that Er,Cr:YSGG laser has a canals decontamination ability when associated to NaOCl irrigation.

Effect of using diode laser on Enterococcus faecalis and its lipoteichoic acid (LTA) in chronic apical periodontitis.

The purpose of this study was to evaluate the effect of diode laser irradiation on Enterococcus faecalis (E. faecalis) and its lipoteichoic acid (LTA). Ninety-six freshly extracted single-rooted teeth were divided into six groups, n = 8 per group. Groups 1, 2, 3, and 4 as laser group (810 nm PILOT™ Diode Laser, 400 µm fiber diameter, continuous mode, 30 s time) with powers at 1.0 W, 1.5 W, 2.0 W, and 2.5 W respectively. Group 5 or positive control group (3 ml of 1% sodium hypochlorite (NaOCl) irrigation) and group 6 or negative control group (3 ml of normal saline (0.9% NaCl) irrigation). Root canal samples were collected before and after receiving laser irradiation and irrigation solution. Cultivable bacteria were determined by counting the colony (CFU/ml). Evaluation of temperature on the external root surface of teeth was done with K type thermocouple using laser at different powers. Enzyme-linked immunosorbant assay (ELISA) was performed to measure the LTA levels and the correlations between E. faecalis count, LTA levels, and rise in temperature were observed using Pearson's correlation test. E. faecalis LTA was subjected to laser irradiation and its structural damage was examined by thin layer chromatography (TLC). Compared with the control groups, all laser groups showed a decreased colony counts and decreased LTA levels with statistically significant difference (p Ë‚ 0.05). The bactericidal effect and LTA reduction of laser was better at 2.5 W power. Laser at 2.5 W power had temperature rise of more than 7 °C which is beyond the safe thermal threshold level. No statistically significant correlation was found between E. faecalis count, levels of LTA, and rise in external root surface temperature (p Ëƒ 0.05). TLC results showed a structural damage in the glycolipid moiety of E. faecalis LTA. Diode laser can effectively reduce the E. faecalis count and its LTA levels.

Ex vivo comparison of antibacterial efficacy of conventional chemomechanical debridement alone and in combination with light-activated disinfection and laser irradiation against Enterococcus faecalis biofilm.

BACKGROUND AND OBJECTIVES: Intracanal disinfection plays an important role in endodontic treatment success. Enterococcus faecalis (E. faecalis) is a resistant microorganism responsible for endodontic infections. We aimed to assess the bactericidal effects of three disinfection methods on E. faecalis biofilm. MATERIALS AND METHODS: Fifty-five freshly extracted single-rooted human teeth were evaluated. A barbed broach was used to extract the pulp tissue. No further root canal preparation was performed. Specimens were sterilized with gamma radiation, and inoculated with E. faecalis suspension. They were then incubated for 4 days and 4 weeks. Biofilm formation was confirmed using a scanning electron microscope (SEM). The teeth were randomly assigned to three subgroups (n = 7) to assess the antimicrobial efficacy of the following three disinfection methods against immature (4-day) and mature (4-week) biofilms: the conventional chemomechanical debridement (CCMD), CCMD + light-activated disinfection (LAD; 810 nm, 0.3 W, 120 J/cm2) with indocyanine Green (EmunDo) as photosensitizer and CCMD + diode laser irradiation (810 nm, 2 W). The teeth were then longitudinally split into two halves and the colony count was reported as colony forming units (CFUs) to assess bacterial viability after each disinfection protocol. RESULTS: None of the disinfection methods could completely remove the biofilm. CCMD + LAD caused the highest and CCMD + diode laser caused the lowest reduction in biofilm. Antibacterial efficacy was significantly lower against the mature (4-week) biofilm compared with immature (4-day) biofilm in all groups (P < 0.05). CONCLUSION: All three disinfection methods were effective for partial elimination of E. faecalis biofilm. But CCMD + LAD was significantly more efficacious in decreasing both mature and immature biofilms.

Antimicrobial Blue Light Inactivation of Microbial Isolates in Biofilms.

BACKGROUND AND OBJECTIVES: Biofilms cause more than 80% of infections in humans, including more than 90% of all chronic wound infections and are extremely resistant to antimicrobials and the immune system. The situation is exacerbated by the fast spreading of antimicrobial resistance, which has become one of the biggest threats to current public health. There is consequently a critical need for the development of alternative therapeutics. Antimicrobial blue light (aBL) is a light-based approach that exhibits intrinsic antimicrobial effect without the involvement of exogenous photosensitizers. In this study, we investigated the antimicrobial effect of this non-antibiotic approach against biofilms formed by microbial isolates of multidrug-resistant bacteria. STUDY DESIGN/MATERIALS AND METHODS: Microbial isolates of Acinetobacter baumannii, Candida albicans, Escherichia coli, Enterococcus faecalis, MRSA, Neisseria gonorrhoeae, Pseudomonas aeruginosa, and Proteus mirabilis were studied. Biofilms were grown in microtiter plates for 24 or 48 hours or in the CDC biofilm reactor for 48 hours and exposed to aBL at 405 nm (60 mW/cm2 , 60 or 30 minutes). The anti-biofilm activity of aBL was measured by viable counts. RESULTS: The biofilms of A. baumannii, N. gonorrhoeae, and P. aeruginosa were the most susceptible to aBL with between 4 and 8 log10 inactivation after 108 J/cm2 (60 mW/cm2 , 30 minutes) or 216 J/cm2 (60 mW/cm2 , 60 minutes) aBL were delivered in the microplates. On the contrary, the biofilms of C. albicans, E. coli, E. faecalis, and P. mirabilis were the least susceptible to aBL inactivation (-0.30, -0.24, -0.84, and -0.68 log10 inactivation, respectively). The same aBL treatment in biofilms developed in the CDC biofilm reactor, caused -1.68 log10 inactivation in A. baumannii and -1.74 and -1.65 log10 inactivation in two different strains of P. aeruginosa. CONCLUSIONS: aBL exhibits potential against pathogenic microorganisms and could help with the significant need for new antimicrobials in clinical practice to manage multidrug-resistant infections. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.

In vitro bactericidal effect of Ho:YAG laser and pneumatic lithotripsy on ureteral stones colonized with Escherichia coli and Enterococcus faecalis.

The endoscopic treatment of urolithiasis has a high success rate and the complications decreased after the development of lithotripsy techniques. The aim of this study is to investigate the in vitro bactericidal effect of laser and pneumatic lithotripsy on urinary stones colonized with Escherichia coli and Enterococcus faecalis. A total of 40 natural calcium oxalate stones, which were obtained from the patients' urinary systems with rigid ureteroscopy were used in the study. Surfaces of the stones were colonized with E. coli and E. faecalis strains. The fragmentation of the stones was performed using holmium:yttrium-aluminum-garnet laser (Ho:YAG laser) and pneumatic lithotripters in vitro in Eppendorf tubes filled with liquid. After fragmentation, samples taken from Eppendorf tubes were inoculated on blood and EMB agar. The number of colonies was evaluated after 18-24-hour incubation period. The laser lithotripsy technique reduced the number of colonies by 100% and had bactericidal effect on E. coli and E. faecalis. Pneumatic lithotripsy technique had no bactericidal effect on these strains (0%). In the fifth minute of laser irradiation, the average temperature in the Eppendorf tube was 51-55 °C, and the average temperature in the tenth minute was 54-60 °C. The temperatures did not change in the fifth and tenth minutes with the pneumatic lithotripsy procedure. The present study revealed the bactericidal effect of Ho:YAG laser on E. coli and E. faecalis in vitro. Increased ambient temperature during Ho:YAG laser lithotripsy is thought to play a role in the bactericidal effect. But the question of whether an ideal lithotripter efficiently inactivates or destroys bacteria has still not been answered in urology practice. This preliminary study showed the bactericidal effect of Ho:YAG laser lithotripsy, but further studies are needed to investigate the bactericidal effect of Ho:YAG laser in vivo.

Near-infrared LEDS provide persistent and increasing protection against E. faecalis.

In a previous study, we have shown that non-coherent light-emitting diodes (LEDs) are able to significantly decrease Enterococcus faecalis viability, in vitro. In particular, the concurrent treatment for 5 min with sodium hypochlorite (NaOCl) 1.0% and LED permitted to obtain the total inactivation of the culture. The primary outcome of this study was to verify if the photoinactivation was only temporary or stable over 1 week. The secondary outcome was to verify if using a minor concentration of NaOCl together with LED irradiation, it was possible to reach the total inactivation of the culture. Bacterial suspensions were subjected to different protocols. NaOCl samples were treated with 0.5% sodium hypochlorite for 5 min; LED 5 were subjected to 880-nm LED irradiation for 5 min and LED+NaOCl were contemporary subjected to LED irradiation and 0.5% sodium hypochlorite for 5 min. Then the colony forming units per milliliter (CFU/mL) were analyzed and recorded at 5 min, 24 h, 48 h and 1 week. The data were submitted to ANOVA and LSD post hoc tests at a level of significance of 0.05. Live dead assay and fluorescent microscope observations were also performed for all group at all timing points. The results showed that the 3 protocol treatments were able to decrease significantly the bacterial count respect controls, however, after 5 min, 24 h and 48 h the most effective reduction was measured for NaOCl and NaOCl+LED 5. In the first 48 h LED 5 alone reduced significantly the bacterial count respect controls, but it was higher respect the other tested groups. On the contrary, after 1 week we found an interesting result: samples treated with LED were characterized by a decrement of bacterial count, independently from the addition of sodium hypochlorite. Samples treated with NaOCl alone were characterized by a mild increment of bacterial count, although results were still significant respect C+. The secondary outcome was not confirmed: a concentration of NaOCl 0.5% together with LED irradiation did not permit to reach the total inactivation of the culture.

Bactericidal Effect of 2780 nm Er,Cr:YSGG Laser Combined with 940 nm Diode Laser in Enterococcus faecalis Elimination: A Comparative Study.

Objective: This study aims to compare the bactericidal efficacy of different Er,Cr:YSGG disinfection methods, particularly combined application of Er,Cr:YSGG and Diode laser. Background: Root canal disinfection is important for long-term endodontic success. Different laser systems are proposed as alternatives to conventional techniques in bacterial elimination. Materials and methods: Ninety-five straight, single-rooted premolars with similar canal dimensions were selected. After the mechanical preparation, the samples were sterilized, and root canals were infected with Enterococcus faecalis. The samples were randomly divided into five groups (n = 15): 5% NaOCl, Er,Cr:YSGG, Er,Cr:YSGG +5% NaOCl, Er,Cr:YSGG + Diode, and control group. Results: The maximal bacterial elimination was observed in Er,Cr:YSGG + NaOCl group (p < 0.005). More bacteria were found in Er,Cr:YSGG and Er,Cr:YSGG + Diode groups compared with 5% NaOCl group (p < 0.005). Marginal significance was detected between the Er,Cr:YSGG group and the Er,Cr:YSGG + Diode group (p = 0.091). Conclusions: The most successful E. faecalis elimination was obtained from laser-activated irrigation group. As combined application of Er,Cr:YSGG and Diode laser gives suggestive results, further studies with larger sample sizes are needed to clarify the outcome.

In Vitro Effect of Er:YAG Laser on Different Single and Mixed Microorganisms Being Associated with Endodontic Infections.

Objective: The purpose of this in vitro study was to evaluate the antimicrobial effect of activated irrigation with different modes of erbium-doped yttrium aluminum garnet (Er:YAG) laser application on microorganisms related to secondary endodontic infection. Background: Er:YAG laser has been recommended as an adjuvant tool for root canal disinfection during endodontic treatment. Materials and methods: Laser-activated irrigation (LAI) with 300 or 600 µm tips were tested with or without intermittent irrigation with 0.9% sodium chloride (NaCl) solution against different microorganisms (five single strains and dual species (Streptococcus gordonii combined with Actinomyces oris or Fusobacterium nucleatum) in root canals after 3 days of incubation. In a 21-day infection model, LAI was used together with intermittent rinsing with sodium hypochlorite (NaOCl) against the dual-species mixtures; here the incidence of microbial regrowth after up to 7 days was monitored. Results: In the 3-day root infection model, LAI protocols did not show any significant reduction of the microbial load when compared with manual irrigation with saline solution. In the 21-day infection, S. gordonii combined with A. oris were not detectable anymore after applying the LAI protocol with a 600 µm tip (30 mJ/10 pps) up to 7 days after treatment. Conclusions: Application of LAI with a 600 µm tip by using an Er:YAG laser might be advantageous in treatment of endodontic infections.

The photo-activated and photo-thermal effect of the 445/970 nm diode laser on the mixed biofilm inside root canals of human teeth in vitro: A pilot study.

AIMS: 1) Evaluation of the photo-thermal (PT) and photo-activated (PAD) antibacterial effect of the 445/970 nm diode laser on E. faecalis, S. aureus and C. albicans mixed biofilms grown together inside root canals of human teeth. 2) Defining a potentially efficient clinical protocol for safe and predictable usage in endodontic procedures. METHODOLOGY: The root canals of 100 extracted human teeth with single straight canals were prepared with ProTaper NEXT files, sterilized, contaminated with a combination of three cultures (E. faecalis, S. aureus, C. albicans) and incubated for 15 days. The samples were randomly distributed into three groups (n = 20) and treated as follows: Group 1 (G1) - the 445 nm photo-thermal (PT) effect, Group 2 (G2) - a combination of the 445 nm and 970 nm PT effect, Group 3 (G3) - the 445 nm photo-activated (PAD) effect with 0.1% riboflavin, Group 4 (G4) - a combination of 3% sodium hypochlorite (NaOCl) and the 445 nm PAD effect. Four samples were used as positive control (non-treated) and four as a negative control. 12 aditional samples were used as a control for the G4 (3% NaOCl rinse without the laser). The number of viable microbes in each canal was determined by the colony forming unit (CFU) count. RESULTS: A statistically significant reduction in the microbial population after all treatments was observed (P < 0.001). Groups 2 and 3 showed similar results, both better than Group 1. Group 4 produced the best results. CONCLUSIONS: The 445 nm PAD protocol has a stronger antimicrobial effect than the 445 nm PT protocol. Prolonged exposure time to laser light and a combination of wavelengths (445/970 PT protocol) helps in the reduction of microbes. C. albicans appears to be more sensitive to laser irradiation than the other bacteria tested in this study. Following current results, tested laser protocols could be recommended for clinical usage but only as an adjunct to "classic" NaOCl rinse since alone they are not able to completely eradicate all microorganisms.

Hybrid UV-C/microfiltration process in membrane photoreactor for wastewater disinfection.

A novel hybrid UV-C/microfiltration process for water disinfection is presented, and its application in continuous mode operation to the removal of different pathogen germs (Escherichia coli, Enterococcus faecalis, and Candida albicans) present in urban wastewater. The membrane photoreactor is based on porous stainless steel membranes coated with a TiO2 layer and illuminated by a UV-C lamp (254 nm). A valve actuator in the outlet of the UV-C stream allows operation of the system under conditions of constant transmembrane pressure (TMP) keeping the UV-C contact time in few seconds, significantly lower than the typical irradiation time employed in TiO2 photocatalytic processes. An E. coli removal of up to 4-log in the permeate stream and up to 2-log in the UV-C outlet was achieved with a 0.2 µm membrane operating with a TMP of 0.5 bar and a UV-C contact time as low as 8 s. The microbial balance data from the cells recovered from the membrane confirmed that 96-98% of the removed microorganisms died due to the UV-C action over the membrane surface. Modification of the membrane with a TiO2 layer has been also shown to be a suitable way to improve both the UV-C inactivation and the filtration efficiency. The results reported in this work constitute a proof of concept of the synergy between UV-C and filtration that can be achieved in a hybrid UV-C/microfiltration system, being a good example of process intensification where two products of different quality can be simultaneously obtained.

Bactericidal efficacy of three parameters of Nd:YAP laser irradiation against Enterococcus faecalis compared with NaOCl irrigation.

The success of endodontic treatment depends on the thorough removal of microorganisms from the root canal system. The search for new ways to eliminate the microorganisms is therefore justified. Nd:YAP is a laser that uses yttrium aluminum perovskite, doped with neodymium crystal, as active laser medium. We used the Nd:YAP laser in an in vitro experiment to evaluate the bactericidal effect of three parameters of Nd:YAP laser-activated irrigation on biofilms of Enterococcus faecalis in root canals. The canals of 45 extracted human single-root teeth were prepared on a #35 Mtwo instrument and contaminated with E. faecalis for 14 days. Forty infected single-root teeth were then randomly divided into four groups according to the irrigation agitation protocols as follows: 5.25% sodium hypochlorite (NaOCl), Nd:YAP laser (180 mJ) + NaOCl, Nd:YAP laser (280 mJ) + NaOCl, and Nd:YAP laser (360 mJ) + NaOCl. The remaining bacteria were counted immediately using the cell count method. Teeth were firstly spilt and one half examined by scanning electron microscopy (SEM). The other half involved examination of bacterial colonization in dentinal tubules using confocal laser scanning microscopy (CLSM). Nd:YAP laser (280 mJ) + NaOCl and Nd:YAP laser (360 mJ) + NaOCl completely removed the E. faecalis biofilms from the root canal walls and made it the cleanest among the treatment groups. Bacterial reductions in the treatment groups for dentinal tubules are presented in a descending order as follows: Nd:YAP laser (360 mJ) (53.7%), Nd:YAP laser (280 mJ) (51.5%) > Nd:YAP laser (180 mJ) (45.3%) > 5.25% NaOCl (31.9%) > control (19.3%) (p < 0.05). Nd:YAP laser of 280 mJ and 360 mJ showed effective bactericidal effect in removing E. faecalis biofilm from the root canal walls and dentinal tubules.

Antimicrobial efficacy of 980 nm diode laser on Enterococcus feacalis in conjunction with various irrigation regimes in infected root canals: An in vitro study.

AIM: The aim of the present study was to evaluate and compare the antimicrobial efficiency of different irrigating solutions against Enterococcus faecalis with or without the use of 980 nm diode laser in infected root canals. MATERIALS AND METHODS: A total of 100 freshly extracted human single-rooted teeth were divided into three experimental groups (n = 30) and one control group (n = 10). Experimental group was subdivided into two subgroups A and B (n = 15), whereas control group was subdivided into positive and negative controls (n = 5). After access cavity preparation, the root canals were prepared using ProTaper rotary instruments. A volume of 5 ml of 5.25% sodium hypochlorite and 17% Ethylenediaminetetraacetic acid, 1.2 ml of 2% chlorhexidine gluconate, and 2 ml of neem solution was used for irrigation in Group I, II, and III, respectively. E. faecalis (ATCC 29212) was inoculated into each canal of experimental groups and incubated for 2 weeks under aerobic conditions at 37°C. In all the subgroups B, laser irradiation was performed, whereas, in subgroup A, no laser irradiation was done. In negative control, E. faecalis was not inoculated in samples, but in positive control, E. faecalis inoculation and incubation procedure was followed. With circumferential filing using #40 reamer, dentinal chips were collected using sterile paper points, vortexed in sterile saline, was then applied to UTI Hicrome agar culture plates and incubated at 37°C for 48 h. The cfu/ml for each plate was calculated using a bacterial colony counter. The data obtained were subjected to statistical analysis using SPSS software version 20.0. RESULTS: The study showed that the least number of bacterial cfu/ml was observed in Group IB. The order of bacterial colony count (cfu/ml) was observed as Group IB

Bactericidal effect of a diode laser on Enterococcus faecalis in human primary teeth-an in vitro study.

BACKGROUND: In recent years, the diode laser (810 nm) has been used for root canal disinfection, which plays an important role in endodontic therapy. This study was undertaken to evaluate the disinfecting ability of a diode laser in experimentally infected root canals of primary teeth. METHODS: Human retained mandibular primary anterior teeth without apical foramen resorption were selected and contaminated with Enterococcus faecalis for 21 days. The specimens were randomly divided into four groups: the negative group (no treatment), positive group (5.25% NaOCl), diode laser group (diode laser), and diode-NaOCl group (diode laser combined with NaOCl). The disinfecting abilities of the treatments were measured by the numbers of bacteria, scanning electron microscopy and confocal laser microscopy (live-dead staining). RESULTS: Eighty teeth were selected. After irradiation and irrigation, the elimination of bacteria and the smear layer in the laser groups and positive group were significantly superior, compared with the negative group (p < 0.01). In the diode-NaOCl group, bacterial reduction reached nearly 100% on the surfaces of root canals; live bacteria were rarely observed, even in deeper dentinal tubules. CONCLUSION: Use of a diode laser, especially in combination with NaOCl, was effective for disinfecting infected root canals of primary teeth.

Antimicrobial Potential of Laser Diode in Infected Dentin.

AIM: To evaluate the antibacterial effect of diode laser, associated or not with 2.5% sodium hypochlorite (NaOCl), against Enterococcus faecalis. MATERIALS AND METHODS: Eighty dentin blocks were obtained from single-rooted human teeth and sterilized. Seventy were inoculated with 0.01 mL of fresh bacterial inoculum (within 24 hours of preparation from pure culture) standardized to 1 McFarland turbidity. Contaminated blocks were incubated for 7 days at 37°C in humid conditions. Ten uncontaminated samples were incubated at 37°C during the contamination period to serve as a negative control group, while 10 of the infected specimens served as a positive control group. The dentin blocks were randomly divided into eight experimental groups (n = 10 each) according to the method of decontamination: 2.5% NaOCl alone; 2.5% NaOCl + photodynamic therapy (PDT) with methylene blue/660 nm laser at 18 J for 180 seconds; 2.5% NaOCl + PDT with methylene blue/660 nm laser at 8 J for 80 seconds; methylene blue alone; PDT alone with methylene blue/660 nm laser at 18 J for 180 seconds; PDT alone with methylene blue/660 nm laser at 8 J laser for 80 seconds; positive control group; and negative control group. Microbial growth was evaluated by culture medium turbidity and microbial concentration was analyzed by UV spectrophotometry (adjusted to read at wavelength l = 600 nM). RESULTS: Root canals treated with laser alone at 18 J for 180 seconds had higher bacterial contamination compared with groups in which NaOCl was used, with or without laser irradiation at 18 J for 180 seconds (p < 0.05). CONCLUSION: Photodynamic therapy with a 660 nm diode laser effectively reduced E. faecalis contamination. These findings can guide development of further studies in search of better alternatives for endodontic treatment. CLINICAL RELEVANCE: Chemical and mechanical root canal preparation plays an essential role in reducing microbial burden. However, microorganisms present in areas not mechanically reachable by endodontic instruments. As an alternative to fix this problem, the laser can be applied.