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.

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.

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.

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.

Effects of prolonged exposure to moderate static magnetic field and its synergistic effects with alkaline pH on Enterococcus faecalis.

Static magnetic field (SMF) has been shown to biologically affect various microorganisms, but its effects on Enterococcus faecalis, which is associated with multiple dental infections, have not been reported yet. Besides, Enterococcus faecalis was found to be resistant to the alkaline environment provided by a major dental antimicrobial, calcium hydroxide. Therefore, the antibacterial activity of prolonged exposure to moderate SMF (170 mT) and its possible synergistic activity with alkaline pH (pH = 9) were evaluated in the study. The ability to form a biofilm under these conditions was examined by crystal violet assay. Real-time quantitative PCR was performed to evaluate the relative expression of stress (dnaK and groEL) and virulence (efaA, ace, gelE and fsrC) related genes. As the results indicated, cell proliferation was inhibited after 120 h of SMF exposure. What's more, the combined treatment of SMF and alkaline pH showed significantly improved antimicrobial action when compared to single SMF and alkaline pH treatment for more than 24 h and 72 h respectively. However, the ability to form a biofilm was also enhanced under SMF and alkaline pH treatments. SMF can induce stress response by up-regulating the expression of dnaK and elevate virulence gene expression (efaA and ace). These responses were more significant and more genes were up-regulated including groEL, gelE and fsrC when exposed to SMF and alkaline pH simultaneously. Hence, combination of SMF and alkaline pH could be a promising disinfection strategy in dental area and other areas associated with Enterococcus faecalis infections.

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.

Determining the optimal dose of 1940-nm thulium fiber laser for assisting the endodontic treatment.

Insufficient cleaning, the complex anatomy of the root canal system, inaccessible accessory canals, and inadequate penetration of irrigants through dentinal tubules minimizes the success of the conventional endodontic treatment. Laser-assisted endodontic treatment enhances the quality of conventional treatment, but each laser wavelength has its own its own limitations. The optimal parameters for the antibacterial efficiency of a new wavelength, 1940-nm Thulium Fiber Laser, were firstly investigated in this study. This paper comprises of two preliminary analyses and one main experimental study, presents data about thermal effects of 1940-nm laser application on root canal tissue, effective sterilization parameters for bacteria, Enterococcus faecalis, and finally the antibacterial effectiveness of this 1940-nm Thulium Fiber Laser irradiation in single root canal. Based on these results, the optimal parameter range for safe laser-assisted root canal treatment was investigated in the main experiments. Comparing the antibacterial effects of four laser powers on an E. faecalis bacteria culture in vitro in 96-well plates showed that the most effective group was the one irradiated with 1 W of laser power (antibacterial effect corresponding to a log kill of 3). After the optimal laser power was determined, varying irradiation durations (15, 30, and 60 s) were compared in disinfecting E. faecalis. Laser application caused significant reduction in colony-forming unit values (CFU) compared with control samples in the 17% ethylenediaminetetraacetic acid (EDTA) group. The results of bacteria counts showed that 1 W with 30 s of irradiation with a 1940-nm thulium fiber laser was the optimal dose for safely achieving maximal bactericidal effect.

Effectiveness of repeated photodynamic therapy in the elimination of intracanal Enterococcus faecalis biofilm: an in vitro study.

The study aimed to investigate the effectiveness of photodynamic therapy in the elimination of intracanal Enterococcus faecalis biofilm and to analyse how a repeated light irradiation, replenishment of oxygen and photosensitiser affect the results of the photodynamic disinfecting protocol. After chemomechanical preparation, 46 single-rooted human teeth were infected with a clinical strain of E. faecalis and incubated for a week in microaerobic conditions. The experimental procedures included groups of single application of photodynamic therapy, two cycles of PDT, irrigation with 5.25% NaOCl solution and negative and positive control. The number of residing bacterial colonies in the root canals was determined based on the CFU/ml method. In the group of preparations irrigated with NaOCl, bacterial colonies were not observed. A single PDT eliminated 45% of the initial CFU/ml. Repeated PDT eradicated 95% of the intracanal bacterial biofilm. Photodynamic therapy has a high potential for the elimination of E. faecalis biofilm. There is a safe therapeutic window where photoinduced disinfection can be used as an adjuvant to conventional endodontic treatment, which remains the most effective.

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.

[Bactericidal effects and smear layer removal of Er:YAG-laser radiation against Enterococcus faecalis in root canals].

OBJECTIVE: To evaluate the cleaning ability of Er:YAG laser against Enterococcus faecalis in root canals.
 Methods: The single-rooted human teeth were sterilized and inoculated with Enterococcus faecalis and were randomly assigned into 2 groups. A group of teeth was irrigated with saline during root canal preparation while another group with NaClO. After mechanical preparation, the two groups were randomly divided into 3 subgroups according to the different treatments: Samples, laser radiation, and calcium hydroxide intracanal medication for 7 days. Bactericical effects were compared among groups. Root canal walls and dental tubules were observed under scanning electron microscope.
 Results: Er:YAG laser was the most efficient way for anti-bacteria among the groups (P<0.05), but there was no significant difference between the 2 laser groups (P>0.05). Meanwhile the smear laser was efficiently removed by laser compared with other treatments, and the laser could open the dentinal tubules.
 Conclusion: Er:YAG laser can be effectively used for root canal disinfection without NaClO and Ca(OH)2.

Effects of salinity and temperature on inactivation and repair potential of Enterococcus faecalis following medium- and low-pressure ultraviolet irradiation.

AIMS: To investigate the medium-pressure (MP) and low-pressure (LP) Ultraviolet (UV) susceptibility and the repair potential of Enterococcus faecalis (DSM 20478) after UV treatment. METHODS AND RESULTS: A range of UV doses from 4 to 19 mJ cm(-2) was selected in this study. Photoreactivation and dark repair performance were investigated under fluorescent light or in the dark respectively. The inactivation and repair performance of UV disinfection under a range of salinities (0, 1%, 3%) and temperature (4 and 25°C) were compared. Results indicated that MP UV exposure resulted in higher inactivation efficiency against Ent. faecalis than LP UV exposure. For repair potential, LP UV resulted in a greater level of light repair than MP UV. Effect of salinity on the inactivation and repair of Ent. faecalis was correlated with UV sources, whereas low temperature generally adversely affected the inactivation efficiency and final repair levels after both MP and LP UV exposure. CONCLUSIONS: Both salinity and temperature demonstrated to play an important role in the inactivation and repair capability when UV light was used to treat ballast water. SIGNIFICANCE AND IMPACT OF THE STUDY: Considering that UV-treated ballast water is exposed or discharged to marine water environment in many countries with various temperature and salinity conditions, results of this study provide significant implications for the management of public health associated with ballast water treatment and discharge.

Photodynamic UVA-riboflavin bacterial elimination in antibiotic-resistant bacteria.

BACKGROUND: To evaluate the bactericidal effect of clinical ultraviolet A (UVA) settings used in photoactivated chromophore for infectious keratitis (PACK)-collagen cross-linking (CXL) in antibiotic-resistant and non-resistant bacterial strains. METHODS: Well-characterized bacterial strains from clinical isolates, without and with antibiotic resistance, were studied in a pairwise comparison. The evaluated pathogens were Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas aeruginosa, and Enterococcus faecalis. Bacteria were dispersed in PBS and diluted to a concentration of approximately 4 × 105 /ml. Riboflavin was added to a concentration of 0.01%. By spreading the solution on a microscope slide, a fluid film layer, with a thickness of around 400 mm, was formed and UVA exposure followed. Eight separate exposures were made for each strain (n = 8). The degree of elimination in resistant and non-resistant pathogens was compared. RESULTS: The bactericidal efficacy of exposure differed between the tested microorganisms, and the mean elimination ranged between 60 and 92%, being most extensive in both of the evaluated Pseudomonas strains and least in the E. faecalis strains. Similar reductions were seen in antibiotic-resistant and non-resistant strains, with the exception of S. aureus, in which the resistant strain metchicillin-resistant Staphylococcus aureus (MRSA) was eradicated in a greater extent than the non-resistant strain (P = 0.030). CONCLUSION: UVA-riboflavin settings used in PACK-CXL are effective in reducing both antibiotic-resistant and non-resistant bacteria. Antibiotic resistance does not appear to be protective against the photooxidative exposure.

The efficacy of photon-initiated photoacoustic streaming and sonic-activated irrigation combined with QMiX solution or sodium hypochlorite against intracanal E. faecalis biofilm.

The aim of the study was to assess the antibacterial efficacy of photon-initiated photoacoustic streaming (PIPS) using an Er:YAG laser and sonic-activated irrigation combined with QMiX irrigant or sodium hypochlorite against Enterococcus faecalis intracanal biofilm. Root canals of 91 human extracted single-canal teeth were instrumented, sterilized, contaminated with E. faecalis and incubated for 15 days. The infected teeth were then randomly distributed into six experimental groups: G1: PIPS/Er:YAG laser (wavelength 2940 nm, pulse energy 20 mJ, 15 Hz, pulse duration 50 µs, energy density 2.06 J/cm(2), 3 × 20 s) with the QMiX irrigant; G2: PIPS/Er:YAG laser-activated 2.5 % NaOCl; G3 sonic-activated irrigation (EndoActivator system) for 60 s with the QMiX irrigant; G4 sonic-activated irrigation for 60 s with 2.5 % NaOCl; G5 30-gauge needle irrigation with the QMiX irrigant; G6 30-gauge needle irrigation with 2.5 % NaOCl. The positive control group was rinsed with sterile saline solution. The root canals were sampled by flushing with saline solution at baseline and after the treatments, serially diluted and cultured. The number of bacteria in each canal was determined by plate count. The presence and the absence of E. faecalis in root canals were demonstrated by polymerase chain reaction (PCR), and the pattern of the bacteria colonization was visualized by scanning electron microscopy. There was significant reduction in the bacterial population for all groups (p < 0.001). The best antibacterial efficacy was recorded after sonic-activated irrigation with both NaOCl (99.999 %) and QMiX (99.999 %) and after PIPS with QMiX (99.999 %), which were more effective than conventional irrigation with NaOCl (99.998 %) and the PIPS with the NaOCl (99.966 %). Also, the PIPS with QMiX solution provided the highest number of sterile samples (five). There was no difference in the bacteria reduction between the active irrigation techniques, regardless of the irrigant used. Although the laser activation did not improve the antimicrobial action of the NaOCl nor QMiX, the fact that it generated the greatest number of sterile samples warrants further investigation.

Growth-dependent photoinactivation kinetics of Enterococcus faecalis.

AIMS: To investigate how the growth stage of Enterococcus faecalis affects its photoinactivation in clear water. METHODS AND RESULTS: Enterococcus faecalis were grown in batch cultures to four different growth stages or grown in chemostats set at four different dilution rates, then harvested and exposed to full spectrum or UVB-blocked simulated sunlight. Experiments were conducted in triplicate in clear water with no added sensitizers. Decay curves were shoulder-log linear and were generally not statistically different in experiments conducted under full spectrum light. Shoulders were longer and first order inactivation rates smaller when experiments were seeded with cells grown to stationary as compared to exponential phase, and for slower growing cells when experiments were done under UVB-blocked light. Chemostat-sourced bacteria generally showed less variability among replicates than batch-sourced cells. CONCLUSIONS: The physiological state of cells and the method via which they are being generated may affect the photoinactivation experimental results. SIGNIFICANCE AND IMPACT OF THE STUDY: Photoinactivation experiments conducted with exponential phase cells may overestimate the photoinactivation kinetics in the environment, particular if UVB-independent mechanisms predominate. Chemostat-sourced cells are likely to provide more consistent experimental results than batch-sourced cells.

Influence of formulation on photoinactivation of bacteria by lumichrome.

Lumichrome, a photodegradation product of riboflavin, is an endogenous compound in humans. The compound is more photostable and a more efficient photogenerator of singlet oxygen than riboflavin. It absorbs radiation in the UVA and blue-light region, which can be an advantage in antibacterial photodynamic therapy (aPDT) of superficial infections. The aim of this study was to investigate the in vitro aPDT effect of various lumichrome pharmaceutical formulations. Solutions of lumichrome (10(-5) - 10(-3)M) were prepared in plain phosphate buffered saline (PBS) or in PBS solutions containing cyclodextrins, DMSO, PEG 400 or polyoxamers (Pluronic). Supersaturated solutions of lumichrome in PBS were prepared via the cosolvent and solvent evaporation method. Phototoxic effects of selected lumichrome preparations were studied in planktonic Gram-positive (E. faecalis) and Gram-negative (E. coli) bacteria models. The UVA/blue light source emitted mainly in the range 340-440 nm. Lumichrome was up to tenfold more phototoxic against Gram-positive than to Gram-negative bacteria. Bacterial eradication was induced after exposure of lumichrome formulations (PBS, PEG 400 and HPγCD) combined with 24J/cm2 UVA/blue light. Increasing the concentration of lumichrome did not enhance the phototoxic effect, probably due to radiation attenuation in the highly absorbing solution (inner filter effect). Cyclodextrins were efficient enhancers of the lumichrome solubility in aqueous solutions, but inhibited the phototoxic effect. The study demonstrates that assuming the use of an optimized formulation, lumichrome has potential as a UVA/blue light photosensitizer in aPDT.

Atomic force microscopy visualization of injuries in Enterococcus faecalis surface caused by Er,Cr:YSGG and diode lasers.

AIM: To visualize by atomic force microscopy the alterations induced on Enterococcus. faecalis surface after treatment with 2 types of laser: Erbium chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser and Diode laser. MATERIAL AND METHODS: Bacterial suspensions from overnight cultures of E. faecalis were irradiated during 30 seconds with the laser-lights at 1 W and 2 W of power, leaving one untreated sample as control. Surface alterations on treated E. faecalis were visualized by atomic force microscopy (AFM) and its surface roughness determined. RESULTS: AFM imaging showed that at high potency of laser both cell morphology and surface roughness resulted altered, and that several cell lysis signs were easily visualized. Surface roughness clearly increase after the treatment with Er,Cr:YSGG at 2W of power, while the other treatments gave similar values of surface roughness. The effect of lasers on bacterial surfaces visualized by AFM revealed drastic alterations. CONCLUSIONS: AFM is a good tool to evaluate surface injuries after laser treatment; and could constitute a measure of antimicrobial effect that can complete data obtained by determination of microbial viability.

Which is the most effective disinfection method in primary root canals: Conventional or newly developed ones?

BACKGROUND: The aim of the study was to evaluate the antimicrobial activity of the-potassium-titanyl-phosphate--the KTP laser and ozone in of primary root canals. MATERIALS AND METHODS: Sixty primary incisor teeth were selected. The specimens were inoculated with 10 mL Enterococcus faecalis (E. faecalis). Groups: The KTP laser (1,5 W); gaseous ozone (150 s); sodium hypochlorite (NaOCl); saline group. Sterile paper points used to sample bacteria from the canals to tubes containing 5 mL of brain heart infusion broth. Then, 10 mL suspension was incubated in culture media for 24 h. Data were analyzed statistically using Kruskal-Wallis and Mann-Whitney U-test. RESULTS: There were statistically significant differences between all groups (P<0.05). Complete sterilization was achieved in the 2.5% NaOCl group. The number of bacteria were significantly reduced in experimental groups in comparison to the saline group. CONCLUSION: The KTP laser and ozone application provided a significant antibacterial effect in primary root canals; however, 2.5% NaOCl was superior.

Are silicone-supported [C60]-fullerenes an alternative to Ru(II) polypyridyls for photodynamic solar water disinfection?

Different photosensitizing materials manufactured by immobilizing (0.5-3.0 g m(-2)) tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) (RDP(2+)), [C60]-fullerene, or 1-(4-methyl)-piperazinylfullerene (MPF) on porous neutral (pSil) or surface-modified anionic (pSil(-)) poly(dimethylsiloxane) are compared on the grounds of their singlet molecular oxygen ((1)O2) production and photodynamic solar water disinfection capability. The C60-based sensitizers display a broad weak absorption in the visible and strong absorption in the UV, while absorption of light by RDP(2+) supported on pSil is strong in both the UV and blue regions. The (1)O2 emission lifetimes (τ(Δ)) determined for RDP(2+) and MPF on porous silicone materials under air are similar (40-50 µs) and correspond to the decay of (1)O2 generated by sensitizers dissolved in the polymer support. In contrast, τ(Δ) measured for C60 in pSil is similar to that observed for MPF or RDP(2+) when immobilized at low loading on pSil, but dramatically increases up to 5 ms if C60 aggregates are formed in the porous material as evidenced by microscopy evaluation. The photosensitizing properties of the dyes, together with their electrical charge and the overall charge of the porous silicone-based materials, lead to highly different sunlight-driven bacteria inactivation efficiencies, as tested with waterborne E. faecalis. RDP/pSil provides efficient disinfection by photosensitization unlike MPF/pSil, which leads to reduced bacteria inactivation rates due to poorer (1)O2 production. C60/pSil and MPF/pSil(-) materials, despite their (1)O2 photogeneration, show unsuccessful waterborne bacteria inactivation due to the negative surface charge of fullerene aggregates in contact with water, and to the net negative charge of the pSil(-), respectively.