Photobiomodulation Reduces the Cytokine Storm Syndrome Associated with COVID-19 in the Zebrafish Model.

Although the exact mechanism of the pathogenesis of coronavirus SARS-CoV-2 (COVID-19) is not fully understood, oxidative stress and the release of pro-inflammatory cytokines have been highlighted as playing a vital role in the pathogenesis of the disease. In this sense, alternative treatments are needed to reduce the level of inflammation caused by COVID-19. Therefore, this study aimed to investigate the potential effect of red photobiomodulation (PBM) as an attractive therapy to downregulate the cytokine storm caused by COVID-19 in a zebrafish model. RT-qPCR analyses and protein-protein interaction prediction among SARS-CoV-2 and Danio rerio proteins showed that recombinant Spike protein (rSpike) was responsible for generating systemic inflammatory processes with significantly increased levels of pro-inflammatory (il1b, il6, tnfa, and nfkbiab), oxidative stress (romo1) and energy metabolism (slc2a1a and coa1) mRNA markers, with a pattern similar to those observed in COVID-19 cases in humans. On the other hand, PBM treatment was able to decrease the mRNA levels of these pro-inflammatory and oxidative stress markers compared with rSpike in various tissues, promoting an anti-inflammatory response. Conversely, PBM promotes cellular and tissue repair of injured tissues and significantly increases the survival rate of rSpike-inoculated individuals. Additionally, metabolomics analysis showed that the most-impacted metabolic pathways between PBM and the rSpike treated groups were related to steroid metabolism, immune system, and lipid metabolism. Together, our findings suggest that the inflammatory process is an incisive feature of COVID-19 and red PBM can be used as a novel therapeutic agent for COVID-19 by regulating the inflammatory response. Nevertheless, the need for more clinical trials remains, and there is a significant gap to overcome before clinical trials can commence.

Effects of photobiomodulation on sport performance in swimming para-athletes - a case series.

This study evaluates photobiomodulation (PBM) on cardio-respiratory function and swimming performance in parathletes. Ten swimming parathletes were tested before PBM, after PBM and placebo irradiation applied on upper musculature. After warmup, the parathletes rested for 1 min, and heart rate was recorded. Three sessions of 50 m free style swimming at maximum effort, with 5-min interval were performed and time, peak and recovery heart rate were recorded. After 1 week, biceps, deltoid, and trapezius received 108 J of energy from an LED array or sham-irradiation in a crossover study. After another week, the same protocol was repeated. All athletes improved time in 50 ms swimming. On average, time decreased 4 s after PBM and 1.5 s after placebo. Also, the peak heartbeat was 10% lower after PBM. A muscular pre-conditioning using PBM with an infrared LED could modulate upper musculature and cardio-respiratory function, leading tobetter swimming performance in parathletes.

Effects of low-intensity laser therapy over mini-implants success rate in pigs.

The success rate of miniscrews when used as temporary orthodontic anchorage is relatively high, but some factors could affect its clinical success such as inflammation around the miniscrew. Low-intensity laser therapy has been widely used for biostimulation of tissue and wound healing specially for its anti-inflammatory effects. The purpose of this study was to evaluate the effect of low-intensity laser therapy over the miniscrew success rate. Five Landrace's pigs received 50 miniscrews on the buccal side of the mandible and on the palate of the maxilla. All the miniscrews were immediately loaded with 250 gf. The laser group were irradiated with a 780-nm diode laser with 70 mWs for 1 min (dose = 34 J/cm(2)); the contralateral side was used as the control group. The miniscrews were photographed and analyzed clinically every week to determine their stability and presence of local inflammation. After 3 weeks, histological analysis and fluorescent microscopy were performed to compare the laser and the control side. Clinical results showed a success rate of 60% for the control group and 80% for the laser-treated group. The histological analysis and fluorescent microscopy demonstrated that the laser group had less inflammatory cells than the control group and the bone neoformation around the miniscrew was more intense. Low-intensity laser therapy increased the success rate of orthodontic miniscrews, probably due to anti-inflammatory effect and bone stimulation.

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.

Plasma proteome responses in zebrafish following λ-carrageenan-Induced inflammation are mediated by PMN leukocytes and correlate highly with their human counterparts.

Regulation of inflammation is a critical process for maintaining physiological homeostasis. The λ-carrageenan (λ-CGN) is a mucopolysaccharide extracted from the cell wall of red algae (Chondrus crispus) capable of inducing acute intestinal inflammation, which is translated into the production of acute phase reactants secreted into the blood circulation. However, the associated mechanisms in vertebrates are not well understood. Here, we investigated the crucial factors behind the inflammatory milieu of λ-CGN-mediated inflammation administered at 0, 1.75, and 3.5% (v/w) by i.p. injection into the peritoneal cavity of adult zebrafish (ZF) (Danio rerio). We found that polymorphonuclear leukocytes (neutrophils) and lymphocytes infiltrating the ZF peritoneal cavity had short-term persistence. Nevertheless, they generate a strong pattern of inflammation that affects systemically and is enough to produce edema in the cavity. Consistent with these findings, cell infiltration, which causes notable tissue changes, resulted in the overexpression of several acute inflammatory markers at the protein level. Using reversed-phase high-performance liquid chromatography followed by a hybrid linear ion-trap mass spectrometry shotgun proteomic approach, we identified 2938 plasma proteins among the animals injected with PBS and 3.5% λ-CGN. First, the bioinformatic analysis revealed the composition of the plasma proteome. Interestingly, 72 commonly expressed proteins were recorded among the treated and control groups, but, surprisingly, 2830 novel proteins were differentially expressed exclusively in the λ-CGN-induced group. Furthermore, from the commonly expressed proteins, compared to the control group 62 proteins got a significant (p < 0.05) upregulation in the λ-CGN-treated group, while the remaining ten proteins were downregulated. Next, we obtained the major protein-protein interaction networks between hub protein clusters in the blood plasma of the λ-CGN induced group. Moreover, to understand the molecular underpinnings of these effects based on the unveiled protein sets, we performed a bioinformatic structural similarity analysis and generated overlapping 3D reconstructions between ZF and humans during acute inflammation. Biological pathway analysis pointed to the activation and abundance of diverse classical immune and acute phase reactants, several catalytic enzymes, and varied proteins supporting the immune response. Together, this information can be used for testing and finding novel pharmacological targets to treat human intestinal inflammatory diseases.

Biofilm retention by 3 methods of ligation on orthodontic brackets: a microbiologic and optical coherence tomography analysis.

INTRODUCTION: The aim of this study was to evaluate biofilm retention around orthodontic brackets related to the method of ligation by using optical coherence tomography (OCT) and microbiologic sampling. METHODS: Seventy-five plastic central incisors for dentures were divided into 3 groups and used with metal brackets with a 0.022-in slot with elastomeric ligature (n = 25), metal brackets with a 0.022-in slot with steel wire ligature (n = 25), and self-ligating brackets with a 0.022-in slot (n = 25). The samples were submersed in a suspension of Streptococcus mutans, genetically engineered to express green fluorescent protein, at 37°C for 72 hours to allow biofilm formation. The samples were then submitted to microbiologic analysis and OCT imaging. RESULTS: The microbiologic analysis and the OCT showed significant differences in biofilm formation depending on the ligating method. Brackets ligated with elastomeric rings held more S mutans biofilm, and steel wire ligation had less biofilm retention compared with the other brackets. CONCLUSIONS: This study provided validation that OCT can be used as a potential qualitative marker of total plaque bacteria that can be rapidly and reliably visualized around orthodontic brackets.

Photodynamic Therapy and Photobiomodulation on Oral Lesion in Patient with Coronavirus Disease 2019: A Case Report.

Objective: This article reports the case of a patient with oral manifestation of coronavirus disease 2019 (COVID-19) treated with photobiomodulation (PBM) and photodynamic therapy (PDT). Background: Some dermatological and oral mucosal lesions have recently been linked to severe acute respiratory syndrome coronavirus 2 infection. Methods: A 65-year-old female patient with a confirmed real-time reverse transcriptase-polymerase chain reaction diagnosis of COVID-19 presented with dry edematous lips, edema with mucosal desquamation, ulceration and blood crust on the inner aspect of the lips, gingival petechiae and erythematous and pseudomembranous lesions on the dorsum of the tongue. The treatment protocol was three sessions of antimicrobial PDT (aPDT) (660 nm diode laser + methylene blue) to the lips and tongue, every 24 h to control contamination, followed by PBM (low-power laser, 100 mW, 2 J/point) to the lips, tongue, and oral mucosa for additional four sessions every 24 h. Results: Therapy association promoted pain control and healing of oral mucosal lesions in 7 days of treatment. Conclusions: PBM and aPDT could be an interesting approach to manage COVID-19 patients.

Effects of antimicrobial photodynamic therapy on antibiotic-resistant Escherichia coli.

This study used Electron Cryo-tomography (ECT) and fluorescent images to evaluate antimicrobial photodynamic therapy (aPDT) on the envelope architecture of a Gram-negative bacteria and the effects of combined therapy of aPDT and antibiotics. Standard and clinical suspension of Escherichia coli were submitted to photodynamic treatment with methylene blue solution (100µM) and a 100 mW LED emitting at 660 nm with 3 and 18 J of energy. As a control group, a suspension of E. coli was submitted to penicillin V for 60 min at 30 °C, to compare the damage in cell wall structure. After treatment, ECT images were collected and E. coli biofilms were grown in glass-cover slides and stained with live/dead staining for fluorescence analysis before and after treatments. Bacteria were also submitted to disc diffusion and MIC50 tests with Ampicillin, Amoxicillin + Clavulanic acid, Clindamycin and Erythromycin. For in vivo experiment Galleria mellonella larvae were infected with E. coli and treated with antibiotics, aPDT or combined therapy. ECT images presented damage to cell walls and vesicles structures inside and outside the bacteria and fluorescent images showed dose dependent effect of aPDT. Antibiotic or aPDT alone did not improve the survival of caterpillars, but the combined therapy significantly increased survival curve. ECT and fluorescent images shows that aPDT seems to promote micro-damages to cell envelope and causes the production of membrane vesicles permeabilizing cell membranes. The results showed that pre-treating bacterial cells with a photosensitizer and light make them more susceptible to antibiotics and could be an alternative to local infection treatment by resistant bacteria.

Effects of maxillary skeletal expansion on respiratory function and sport performance in a para-athlete - A case report.

The aim of this case report was to demonstrate the effects of the Maxillary Skeletal Expander (MSE) used to orthopedically correct a maxillary constriction, on the respiratory functions and swimming performance of a Para-athlete. Cone-beam computed tomography (CBCT) images taken before and after MSE activation were used to demonstrate the disarticulation of midpalatal suture, and the changes involved in dental and nasomaxillary structures, nasal cavity and pharyngeal airway. Respiratory tests included: maximum inspiratory and expiratory pressure, oral peak expiratory flow and inspiratory nasal flow. The 6-min-walk and heart rate recovery tests were also performed. Patient's swimming performances during national swimming competitions were compared. CBCT images showed that palatal expansion was 5.91  mm at the suture, and that nasal and pharyngeal airways increased in volume by 31%. All respiratory indices improved after MSE activation. The 6-min walk test and heart rate recovery test performance also improved after the maxillary expansion. Patient's swimming performance in all category were anemic prior to the treatment, but performance improved considerably after the expansion, particularly the 100 m-Backstroke modality. MSE treatment had a significant positive impact in respiratory functions and sport performance.

The potential of commercially available phytotherapeutic compounds as new photosensitizers for dental antimicrobial PDT: A photochemical and photobiological in vitro study.

The present study evaluated the effectiveness of extracts of commercially available Curcuma longa, Citrus lemon, Hamamelis virginiana and Hypericum perforatum as photosensitizers in Antimicrobial Photodynamic Therapy (aPDT). Each photosensitizer (PS) was analyzed in a spectrophotometer between 350 and 750 nm to determine the ideal light source. Once the absorption bands were determined, three light sources were selected. To determine the concentration of use, the compounds were tested at different concentrations on bovine dentin samples to evaluate the risk of staining. Once the concentration was determined, the PSs were evaluated for dark toxicity and phototoxicity on fibroblast and bacteria culture. Each compound was then irradiated with each light source and evaluated for the production of reactive oxygen species (ROS). The bacterial reduction was tested on E. faecalis culture in planktonic form and on biofilm using an energy of 10 J and an Energy Density of 26 J/cm2. The tested compounds exhibited light absorption in three bands of the visible spectrum: violet (405 nm), blue (460 nm) and red (660 nm). At a 1:6 concentration, none of the compounds caused tooth staining as they did not exhibit significant toxicity in the cells or bacterial suspension. Additionally, significant ROS production was observed when the compounds were irradiated at each wavelength. When aPDT was performed on the plactonic and biofilm bacteria, significant microbial reduction was observed in both cases, reaching a reduction of up to 5Logs. In conclusion, extracts of Curcuma longa, Citrus lemon, Hamamelis virginiana and Hypericum perforatum exhibited potential for use as photosensitizing agents in aPDT.

Photobiomodulation increases intrusion tooth movement and modulates IL-6, IL-8 and IL-1ß expression during orthodontically bone remodeling.

This study investigated the effects of photobiomodulation (PBM) on upper molar intrusion movement, regarding acceleration of orthodontic movement and its molecular effects. The sample consisted of 30 patients with indication of tooth intrusion for oral rehabilitation. Teeth were divided into three different groups: G1 (n = 10) pre-molars without force or laser application (control); G2 (n = 10) upper molar intrusion; and G3 (n = 10) upper molar intrusion and PBM. On PBM treated molars, the teeth were irradiated with a low-power diode laser (808 nm, 100 mW), receiving 1 J per point, density of 25 J/cm2 , with application of 10 s per point, 10 points (5 per vestibular and 5 per palatal region). Orthodontic force of intrusion applied every 30 days and PBM was performed immediately, 3 and 7 days after force application for 3 months. Gingival crevicular fluid was collected at the same time periods as the laser applications and interleukins (IL) 1-ß, -6 and -8 were evaluated by enzyme-linked immunosorbent assay. Clinical measures were performed monthly to verify the amount of intrusion. The levels of IL-6, IL-8 and IL-1ß increased under orthodontic force (G2 and G3) when compared to control group (G1), however, the cytokines levels were significantly higher after PBM (G3). The mean intrusion velocity was 0.26 mm/month in the irradiated group (G3), average duration of 8 months vs 0.17 mm/month for the non-irradiated group (G2), average duration of 12 months. This study suggests that PBM accelerates tooth movement during molar intrusion, due to modulation of IL-6, IL-8 and IL-1ß during bone remodeling.

Photoelastic analysis of tension distribution in different orthodontic approaches for closing anterior open bites.

BACKGROUND: This study aims to evaluate tension distribution in the anterior region of the mandible during two orthodontic mechanical approaches to treat anterior open bite. METHODS: It was an in-vitro experimental study, that analyzed 5 models of photoelastic resin, simulating a lower dental arch. The anterior teeth underwent orthodontic forces of the Blue Elgiloy® 0.016" x 0.022" (MEAW technique) and Gummetal® 0.018"x 0.022" (GEAW technique) archwires. Tension distribution was assessed on three different points in the lower dental midline and was measured using a reflection polariscope. The archwires were evaluated with and without anterior elastic bands (6 oz, 170 g) installed between lateral incisors and lower canines, as recommended for anterior open bite. RESULTS: The highest magnitudes of tension generated by the archwires were observed in the cervical regions of the teeth (on average 50% higher than those at the most apical point) regardless of the technique used. Comparatively, the GEAW technique showed significantly (P<0.05) lower tension values (16 MPa) than the MEAW technique using Blue Elgiloy (24 MPa). CONCLUSIONS: The GEAW technique showed a more favorable tension distribution pattern than the MEAW approach. The use of elastic bands improved tension distribution, regardless of the technique. The results also suggest that the use of anterior elastic bands in both techniques reduced the tensions released by the intrusive forces generated by the archwires.

Effects of Photobiomodulation on SOFAT, A T-cell-derived Cytokine, May Explain Accelerated Orthodontic Tooth Movement.

Orthodontic tooth movement is based on mechanical forces inducing bone remodeling, and several methods have been proposed to increase tooth movement, including photobiomodulation. This study evaluated, in an animal model, the effects of photobiomodulation on SOFAT-a secreted osteoclastogenic factor of activated T cells and RANK-L during tooth movement. The results showed that tooth displacement, RANK-L and SOFAT levels were significantly greater compared to Control group. SOFAT may play an important role in bone remodeling during orthodontic movement, possibly increasing the osteoclast cells at the compression area and bone remodeling activity.

Methylene Blue and Hydrogen Peroxide for Photodynamic Inactivation in Root Canal - A New Protocol for Use in Endodontics.

OBJECTIVE: Antimicrobial photodynamic therapy (aPDT) is a controversial approach for endodontic disinfection. The objective of this study was to test the photosensitiser (PS) concentration and assess the optical shielding phenomenon, the use of hydrogen peroxide (H2O2) and minimal energy irradiation to optimise endodontic aPDT for suggesting a protocol for clinical use. METHODS: Different parameters for aPDT were tested. Aqueous solutions of methylene blue (MB) at 50, 100, 150 and 300 µM were tested in vitro for optical shield and reactive oxygen species (ROS) production by the reduction of N,N-dimethyl-4-notrosoaniline (RNO) at 440 nm absorbance when irradiated using a diode laser (660 nm). Ten single-rooted teeth were inoculated with bioluminescent bacteria Pseudomonas aeruginosa for 72 hours to form biofilms. Bioluminescence imaging was used to serially evaluate the minimum energy necessary during endodontic aPDT using MB and a diode laser coupled to an optical fibre for intracanal microbial reduction. In addition, teeth (n=21) infected with Enterococcus faecalis were treated with sequential combinations of endodontic aPDT and H2O2 and the colony-forming unit (CFU) was determined. RESULTS: ROS production was inversely proportional to the MB concentration in the solution due to quenching of MB. Optical shielding limited light penetration at high MB concentrations. The use of H2O2 before aPDT achieved higher disinfection compared to conventional aPDT or when MB was irradiated in an H2O2 solution. Energy irradiation of 9.6 J achieved a significant reduction and further light delivery did not produce further reduction. CONCLUSION: PS concentration of about 50 µM, biofilm pre-treatment with H2O2 for 1 min and energy irradiation around 10 J appear to be an effective protocol for endodontic aPDT.

Methylene Blue and Hydrogen Peroxide for Photodynamic Inactivation in Root Canal - A New Protocol for Use in Endodontics.

OBJECTIVE: Antimicrobial photodynamic therapy (aPDT) is a controversial approach for endodontic disinfection. The objective of this study was to test the photosensitiser (PS) concentration and assess the optical shielding phenomenon, the use of hydrogen peroxide (H2O2) and minimal energy irradiation to optimise endodontic aPDT for suggesting a protocol for clinical use. METHODS: Different parameters for aPDT were tested. Aqueous solutions of methylene blue (MB) at 50, 100, 150 and 300 µM were tested in vitro for optical shield and reactive oxygen species (ROS) production by the reduction of N,N-dimethyl-4-notrosoaniline (RNO) at 440 nm absorbance when irradiated using a diode laser (660 nm). Ten single-rooted teeth were inoculated with bioluminescent bacteria Pseudomonas aeruginosa for 72 hours to form biofilms. Bioluminescence imaging was used to serially evaluate the minimum energy necessary during endodontic aPDT using MB and a diode laser coupled to an optical fibre for intracanal microbial reduction. In addition, teeth (n=21) infected with Enterococcus faecalis were treated with sequential combinations of endodontic aPDT and H2O2 and the colony-forming unit (CFU) was determined. RESULTS: ROS production was inversely proportional to the MB concentration in the solution due to quenching of MB. Optical shielding limited light penetration at high MB concentrations. The use of H2O2 before aPDT achieved higher disinfection compared to conventional aPDT or when MB was irradiated in an H2O2 solution. Energy irradiation of 9.6 J achieved a significant reduction and further light delivery did not produce further reduction. CONCLUSION: PS concentration of about 50 µM, biofilm pre-treatment with H2O2 for 1 min and energy irradiation around 10 J appear to be an effective protocol for endodontic aPDT.

Effects of Low-Level Laser Therapy in Orthodontic Patients on Immediate Inflammatory Response After Mini-Implants Insertion: A Preliminary Report.

BACKGROUND: The primary stability of a mini-implant is crucial to treatment sequence since most orthodontic mini-implant failures occur at an early stage. Irritation or inflammation of peri-implant tissues has been related to decreasing mini-implant success. PURPOSE: This study evaluates the effect of low-level laser therapy on initial inflammation after orthodontic mini-implants installation. METHODS: Ten volunteers received two mini-implants (1.3 mm diameter, 7 mm length). One mini-implant was inserted on each side of the maxilla following manufacturer recommendation. On the right side, low-level laser therapy (LLLT) was applied (diode laser 660 nm, 40 mW, 1 min, 2.4 J of total energy). Peri-implant crevicular fluid (PGF) was obtained after 24 h (T1), 48 h (T2), and 72 h (T3) to identify levels of interleukin (IL)-6 and IL-8 around mini-implants and around upper first premolars. RESULTS: An increase in interleukin levels was observed for both groups, compared to upper first premolar. PGF around nonirradiated mini-implants showed higher levels of IL-8. Levels of IL-6 24 h after mini-implant insertion were higher for laser group. CONCLUSIONS: LLLT modulates the initial inflammation after the insertion of mini-implant, possibly increasing the mini-implant success prognostic and decreasing patient discomfort.

Effects of antimicrobial photodynamic therapy and surgical endodontic treatment on the bacterial load reduction and periapical lesion healing. Three years follow up.

Besides the advances in endodontics, there are situations in which surgery is necessary to retain a tooth that otherwise would be extracted. This study analyzes the microbial reduction after conventional periapical surgery followed by antimicrobial photodynamic therapy (aPDT) in 3 years follows up. Twenty-eight teeth needing periapical surgery were enrolled in this study. Microbiological samples were taken after: (1) accessing the lesion, (2) surgical procedure, and (3) aPDT. The teeth received a full mucoperiosteal flap, osteotomy with a high-speed bur, manual curettage of lesion and of the root, root-end resection and retrograde cavities prepared using ultrasonic retro-tips. After the conventional procedure the cavities received an aqueous solution of methylene blue (60 µM, 3 min) and were irradiated with a diode laser λ=660 nm (6 min, 15 J). After aPDT, a retrograde filling with mineral trioxide aggregate, flap re-positioning, and sutures conventionally ended the procedure. In all the cases, a periapical X-ray exam was taken before and after the surgical procedure. The microbiological samples showed an overall significant reduction, surgical procedure achieved a mean reduction of about 3.5 log while aPDT achieved a 5 log reduction. After 36 months of follow up the patients had, in average a reduction of 78% of the periapical lesion area. It appears that surgical endodontic treatment associated with antimicrobial photodynamic therapy highly improves the microbial reduction compared to the traditional technique and this could directly affect the treatment prognosis and periapical lesion healing.

Effects of photodynamic therapy on Gram-positive and Gram-negative bacterial biofilms by bioluminescence imaging and scanning electron microscopic analysis.

OBJECTIVE: The aim of this study was to test photodynamic therapy (PDT) as an alternative approach to biofilm disruption on dental hard tissue, We evaluated the effect of methylene blue and a 660 nm diode laser on the viability and architecture of Gram-positive and Gram-negative bacterial biofilms. MATERIALS AND METHODS: Ten human teeth were inoculated with bioluminescent Pseudomonas aeruginosa or Enterococcus faecalis to form 3 day biofilms in prepared root canals. Bioluminescence imaging was used to serially quantify and evaluate the bacterial viability, and scanning electron microscopic (SEM) imaging was used to assess architecture and morphology of bacterial biofilm before and after PDT employing methylene blue and 40 mW, 660 nm diode laser light delivered into the root canal via a 300 µm fiber for 240 sec, resulting in a total energy of 9.6 J. The data were statistically analyzed with analysis of variance (ANOVA) followed by Tukey test. RESULTS: The bacterial reduction showed a dose dependence; as the light energy increased, the bioluminescence decreased in both planktonic suspension and in biofilms. The SEM analysis showed a significant reduction of biofilm on the surface. PDT promoted disruption of the biofilm and the number of adherent bacteria was reduced. CONCLUSIONS: The photodynamic effect seems to disrupt the biofilm by acting both on bacterial cells and on the extracellular matrix.