Evaluation of antimicrobial photodynamic therapy and minimal intervention associated with deproteinisation in permanent teeth with molar incisor hypomineralisation: study protocol for a clinical, controlled, blinded trial.

INTRODUCTION: Molar incisor hypomineralisation (MIH) is a qualitative defect of enamel development that occurs in the mineralisation phase. MIH affects one or more permanent molars and, occasionally, permanent incisors. The aim of the proposed study is to evaluate the clinical effect of antimicrobial photodynamic therapy (aPDT) on permanent teeth with MIH through decontamination and sensitivity control. METHODS AND ANALYSIS: Patients from 8 to 12 years of age with permanent molars will be randomly allocated to three groups. Group 1: selective chemical-mechanical removal of carious dentinal tissue around the walls of the cavity with Papacárie Duo and a curette followed by the application of aPDT and deproteinisation with Papacárie Duo; group 2: selective removal of carious dentinal tissue around the walls of the cavity with a curette, followed by the application of aPDT and deproteinisation with a 5% sodium hypochlorite solution; group 3: selective removal of carious dentinal tissue using a curette. The selected teeth must have a carious lesion in the dentin and posteruptive enamel breakdown on one or more surfaces with an indication for clinical restorative treatment. The teeth will subsequently be restored using a mixed technique with resin-modified glass ionomer cement and bulk-fill composite resin. The data will be submitted to descriptive statistical analysis. Associations with age and sex will be tested using either the χ2 test or Fisher's exact test. Pearson's correlation coefficients will be calculated to determine the strength of correlations between variables. Comparisons of the microbiological results (colony-forming units) will be performed using analysis of variance and the Kruskal-Wallis test. Kaplan-Meier survival analysis will be performed to assess the performance of the restorations. ETHICS AND DISSEMINATION: This protocol has been approved by the Human Research Ethics Committee of Nove de Julho University (certificate number: 61027522.0.0000.5511/approval date: 23 August 2022). The findings will be published in a peer-reviewed journal. TRIAL REGISTRATION NUMBER: NCT05443035.

The influence of growth medium on the photodynamic susceptibility of Aggregatibacter actinomycetemcomitans to antimicrobial blue light.

The aim of this study was to investigate whether antimicrobial blue light (aBL) can cause the death of Aggregatibacter actinomycetemcomitans (A.a) and to determine the influence of different culture media, specifically brain heart infusion and blood agar, on bacterial survival fraction. An LED emitting at 403 ± 15 nm, with a radiant power of 1W, irradiance of 588.2 mW/cm2, and an irradiation time of 0 min, 1 min, 5 min, 10 min, 30 min, and 60 min, was used. The plates were incubated in microaerophilic conditions at 37 °C for 48 h, and the colony-forming units were counted. The photosensitizers were investigated using spectroscopy and fluorescence microscopy. There was no significant difference between the culture media (p > 0.05). However, a statistical reduction in both media was observed at 30 min (1058 J/cm2) (p < 0.05). The findings of this study suggest that aBL has the potential to kill bacteria regardless of the culture media used. Light therapy could be a promising and cost-effective strategy for preventing periodontal disease when used in combination with mechanical plaque control.

Comparative study between photodynamic therapy and the use of probiotics in the reduction of halitosis in mouth breathing children: Study protocol for a randomized controlled clinical trial.

INTRODUCTION: Halitosis is a term that defines any odor or foul smell the emanates from the oral cavity, the origin of which may be local or systemic. One of the causes of local or oral halitosis is low salivary flow and dry mouth, which is also one of the complaints of individuals with the mouth-breathing habit. The aim of this study is to determine the effectiveness of antimicrobial photodynamic therapy (aPDT) and the use of probiotics for the treatment of halitosis in mouth-breathing children. METHODS: Fifty-two children between 7 and 12 years of age with a diagnosis of mouth breathing and halitosis determined through an interview and clinical examination will be selected. The participants will be divided into 4 groups: Group 1-treatment with brushing, dental floss and tongue scraper; Group 2-brushing, dental floss and aPDT applied to the dorsum and middle third of the tongue; Group 3-brushing, dental floss and probiotics; Group 4-brushing, dental floss, aPDT and probiotics. The use of a breath meter and microbiological analysis of the tongue coating will be performed before, immediately after treatment and 7 days after treatment. The quantitative analysis will involve counts of colony-forming bacteria per milliliter and real-time polymerase chain reaction. The normality of the data will be determined using the Shapiro-Wilk test. Parametric data will be submitted to analysis of variance and nonparametric data will be compared using the Kruskal-Wallis test. The results of each treatment in the different periods of the study will be compared using the Wilcoxon test. DISCUSSION: Due to the low level of evidence, studies are needed to determine whether treatment with aPDT using annatto as the photosensitizer and blue led as the light source is effective at diminishing halitosis in mouth-breathing children.

Antimicrobial photodynamic therapy mediated by methylene blue in surfactant vehicle as adjuvant to periodontal treatment. Randomized, controlled, double-blind clinical trial.

BACKGROUND: Antimicrobial photodymanic therapy mediated by methylene blue has been investigated as an adjunctive to periodontal treatment but the dimerization of photosensitizer molecules reduces the phototoxic effects. Sodium dodecyl sulfate is a surfactant that may control this aggregation. The aim of this study was evaluated the photodynamic effect of methylene blue in sodium dodecyl sulfate in periodontitis. METHODS: 36 participants with periodontitis were selected and allocated randomly in two group for intervention and other two for control - all of them were treated with scaling and root planing before aPDT. Three periodontal evaluations were done: at the selection time, at the day of intervention and thirty-day after this. Pre-irradiation time was 1 min and 2 min for irradiation. Laser (Therapy XT, DMC, São Carlos, Brazil) with wavelength of 660 nm and 100 mW of power was used. Two photosensitizer solutions with 100 µM methylene blue was used, one of them was in water and other in 0,25% of sodium dodecyl sulfate. Two sites of each participant were selected for the experimental procedures. Microbiological evaluations were performed to quantify microorganisms before and immediately after intervention. Quantitative microbiological evaluation was the primary outcome; morphological aspects of bacterial colony, and clinical probing depth was the secondary one. RESULTS: There was no significant difference between the groups in both bacterial reduction and the clinical parameter evaluated. CONCLUSION: The effect of methylene blue in surfactant did not cause enough phototoxic effects that could promote reduction of periodontal pocket depth.

Antimicrobial Photodynamic Therapy in the Nasal Decolonization of Maintenance Hemodialysis Patients: A Pilot Randomized Trial.

RATIONALE & OBJECTIVE: Infections are an important cause of mortality among patients receiving maintenance hemodialysis. Staphylococcus aureus is a frequent etiological agent, and previous nasal colonization is a risk factor for infection. Repeated antimicrobial decolonization reduces infection in this population but can induce antibiotic resistance. We compared photodynamic therapy, a promising bactericidal treatment that does not induce resistance, to mupirocin treatment among nasal carriers of S aureus. STUDY DESIGN: Randomized controlled pilot study. SETTING & PARTICIPANTS: 34 patients receiving maintenance hemodialysis who had nasal carriage of S aureus. INTERVENTIONS: Patients were randomly assigned to decolonization with a single application of photodynamic therapy (wavelength of 660nm, 400mW/cm2, 300 seconds, methylene blue 0.01%) or with a topical mupirocin regimen (twice a day for 5 days). OUTCOME: Nasal swabs were collected at time 0 (when the carrier state was identified), directly after treatment completion, 1 month after treatment, and 3 months after treatment. Bacterial isolates were subjected to proteomic analysis to identify the species present, and antimicrobial susceptibility was characterized. RESULTS: All 17 participants randomized to photodynamic therapy and 13 of 17 (77%) randomized to mupirocin were adherent to treatment. Directly after treatment was completed, 12 participants receiving photodynamic therapy (71%) and 13 participants treated with mupirocin (77%) had cultures that were negative for S aureus (risk ratio, 0.92 [95% CI, 0.61-1.38]; P=0.9). Of the patients who had negative cultures directly after completion of photodynamic therapy, 67% were recolonized within 3 months. There were no adverse events in the photodynamic therapy group. LIMITATIONS: Testing was restricted to assessing nasal colonization; infectious complications were not assessed. CONCLUSIONS: Photodynamic therapy is a feasible approach to treating nasal carriage of S aureus. Future larger studies should be conducted to determine whether photodynamic therapy is equivalent to the standard of care with mupirocin. FUNDING: Government grant (National Council for Scientific and Technological Development process 3146682020-9). TRIAL REGISTRATION: Registered at ClinicalTrials.gov with study number NCT04047914.

The importance of combining methods to assess Candida albicans biofilms following photodynamic inactivation.

BACKGROUND: Methylene blue (MB)-mediated photodynamic inactivation (PDI) has shown good results in killing Candida spp. Although MB solutions are commonly used, new formulations have been designed to improve PDI. However, chemical substances in the formulation may interfere with the PDI outcome. In this sense, different methodologies should be used to evaluate PDI in vitro. Herein, we report different methodologies to evaluate the effects of PDI with an oral formulation (OF) containing 0.005% MB on Candida albicans biofilm. METHODS: Biofilms were treated using the MB-OF, with 5 min pre-irradiation time and exposure to a 640 nm LED device (4.7 J/cm2). PDI was evaluated by the XTT reduction test, counting the colony forming units (CFU), a filamentation assay, crystal violet (CV) staining, and scanning electronic microscopy (SEM). RESULTS: PDI was able to reduce around 1.5 log10 CFU/mL, even though no significant differences were noted in metabolic activity in comparison to the control immediately after PDI. A significant decrease in yeast to hyphae transition was observed after PDI, while the biofilm exhibited flattened cells and a reduced number of yeasts in SEM. The CV assay showed increased biomass. CONCLUSION: MB-OF-mediated PDI was effective in C. albicans biofilms, as it significantly reduced the CFU/mL and the virulence of surviving cells. The CV data were inconclusive, since the OF components interacted with the CV, making the data useless. Taken together, our data suggest that the association of different methods allows complementary responses to assess how PDI mediated by a formulation impacts biofilms.

A common oral pathogen Porphyromonas gingivalis induces myocarditis in rats.

AIM: To evaluate whether Porphyromonas gingivalis (P. gingivalis) inoculation could induce cardiac remodelling in rats. MATERIALS AND METHODS: The study was conducted on 33 Wistar rats, which were distributed in the following experimental groups: not inoculated; inoculated with 1 × 108 CFU/ml of bacteria; inoculated with 3 × 108 CFU/ml of bacteria. The animals were inoculated at baseline and on the 15th day of follow-up. Blood collection was performed at baseline and 60 min after each inoculation. At 29 days, the animals were subjected to echocardiography and at 30 days to haemodynamic studies before sacrificing them. RESULTS: Impact of the bacteria was more evident in rats that received higher P. gingivalis concentration. Thus, 3 × 108 CFU/ml of bacteria increased the rectal temperature and water content in the lung as well as myocardial necrosis and fibrosis. P. gingivalis induced the intensification of DNA fragmentation and increased the levels of malondialdehyde, oxidized proteins, and macrophage expression in the myocardium. These findings were associated with lower LV isovolumetric relaxation time, +dP/dt, -dP/dt, and higher end-diastolic pressure. CONCLUSIONS: P. gingivalis bacteraemia is significantly associated with adverse cardiac remodelling and may play a biological role in the genesis of heart failure.

Photobiomodulation therapy preconditioning modifies nitric oxide pathway and oxidative stress in human-induced pluripotent stem cell-derived ventricular cardiomyocytes treated with doxorubicin.

Doxorubicin (DOX) is an anthracycline antibiotic that exhibits high heart toxicity. Human-induced pluripotent stem cell-derived ventricular cardiomyocytes (hiPSC-vCMs) are important in vitro models for testing drug cardiotoxicity. Photobiomodulation therapy (PBMT) is a non-invasive therapy that stimulates cells growth and self-repair using light irradiation. This study aimed to investigate the in vitro effects of PBMT preconditioning on cardiotoxicity induced by DOX. HiPSC-vCMs were treated with PBMT for 500 s, followed by the addition of 2 µM DOX. LED irradiation preconditioning parameters were at 660 nm with an irradiance of 10 mW/cm2, performing 5 J/cm2, followed by 24-h DOX exposure (2 µM). Human iPSC-vCMs treated with 2 µM DOX or irradiated with PBMT composed the second and third groups, respectively. The control group did neither receive PBMT preconditioning nor DOX and was irradiated with a white standard lamp. Cells from all groups were collected to perform mRNA and miRNA expressions quantification. PBMT, when applied before the DOX challenge, restored the viability of hiPSC-vCMs and reduced ROS levels. Although downregulated by DOX, myocardial UCP2 mRNA expression presented marked upregulation after PBMT preconditioning. Expression of eNOS and UCP2 mRNA and NO production were decreased after DOX exposure, and PBMT preconditioning before the DOX challenge reversed these changes. Moreover, our data indicated that PBMT preconditioning lowered the miR-24 expression. Our data suggested that PBMT preconditioning ameliorated in vitro DOX-induced cardiotoxicity on transcription level, restoring NO levels and reducing oxidative stress.

Effect of antimicrobial photodynamic therapy with red led and methylene blue on the reduction of halitosis: controlled microbiological clinical trial.

To determine the effect of antimicrobial photodynamic therapy (aPDT) using a red light-emitting diode (LED) on the reduction of halitosis and microbiological levels in the tongue coating immediately after irradiation, 7, 14, and 30 days after treatment. Forty-five young adults diagnosed with halitosis were allocated to three groups: G1, aPDT with 0.005% methylene blue and red LED (660 nm, four irradiation points, 90 s per point, power of 400 mW, 36 J per point, radiant exposure of 95 J/cm2, continuous wave); G2, tongue scraping; and G3, tongue scraping and aPDT. Gas chromatography was performed before and immediately after treatment, as well as at the different follow-up times. Microbiological samples were collected at the same times from the dorsum of the tongue, and bacteria were quantified in the samples using real-time PCRq. The Wilcoxon test was used for the intragroup analyses, and the Kruskal-Wallis test was used for the intergroup analyses. In the intragroup analyses, differences were found before and immediately after treatment in all groups (p < 0.05). The effect was maintained after 7 days only in the tongue scraping group (p < 0.05). In the intergroup analysis, no statistically significant differences were found among the groups (p > 0.05). For the microbiological analyses, no statistically significant differences were found in the groups/bacteria that were analyzed (p > 0.05). aPDT using a red LED and 0.005% methylene blue caused an immediate reduction in halitosis, but the effect was not maintained after 7, 14, or 30 days. No reduction occurred in the number of bacteria investigated or the quantification of universal 16S rRNA. ClinicalTrials.gov Identifier: NCT03656419.

Erythrosine as a photosensitizer for antimicrobial photodynamic therapy with blue light-emitting diodes - An in vitro study.

BACKGROUND: This study aims to test the absorbance of a new composition of erythrosine, its pH, cell viability and potential as a photo sensitizer against Candida albicans when irratiaded with blue light emitting-diode (LED). METHODS: For pH and absorbance tests, erythrosine was prepared at a concentration of 0.03/ml. The cells of the L929 strain were cultured and the alamarBlue® assay was performed on samples to assess cell viability. For the microbiological essay, the strain of Candida albicans ATCC 90028 was selected. Yeast suspensions were divided into the following groups: control without irradiation or photosensitizer (C), irradiated group without photosensitizer (L), photosensitizer group without irradiation (0), and groups that received photosensitizer and irradiation, called aPDT groups. RESULTS: Erythrosine had no significant changes in pH and its absorbance was also consistent (≅400 nm). When it came to cell viability, on the first day, the group that was in contact with the dye and irradiated with the LED in minimun power was found to have the higher cell proliferation. On day 3, both irradiated groups (maximum and minimum) showed the highest cell proliferation. In the microbiological essay with C. albicans, aPDT groups started to show microbial reduction after 60 and 90 s of irradiation and when irradiated for 120 s, 6 microbial reduction logs were found. CONCLUSIONS: The erythrosine in question is a PS, with pH stability, blue light absorbance, cell viability and efficacy against C. albicans. More studies with this PS should be encouraged in order to verify its performance in aPDT.

Comparative study between photodynamic therapy with urucum + Led and probiotics in halitosis reduction-protocol for a controlled clinical trial.

BACKGROUND: Halitosis is a term that defines any foul odor emanating from the oral cavity. The origin may be local or systemic. The aim of the proposed protocol is to determine whether treatment with antimicrobial photodynamic therapy (aPDT) and treatment with probiotics are effective at eliminating halitosis. MATERIALS AND METHODS: Eighty-eight patients, from 18 to 25 years old with a diagnosis of halitosis (H2S≥112 ppb, determined by gas chromatography) will be randomly allocated to four groups (n = 22) that will receive different treatments: Group 1 -treatment with teeth brushing, dental floss and tongue scraper; Group 2 -brushing, dental floss and aPDT; Group 3 -brushing, dental floss and probiotics; Group 4 -brushing, flossing, aPDT and probiotics. The results of the halimetry will be compared before, immediately after, seven days and thirty days after treatment. The microbiological analysis of the coated tongue will be performed at these same times. The normality of the data will be determined using the Shapiro-Wilk test. Data with normal distribution will be analyzed using analysis of variance (ANOVA). Non-parametric data will be analyzed using the Kruskal-Wallis test. The Wilcoxon test will be used to analyze the results of each treatment at the different evaluation periods. CLINICAL TRAIL REGISTRATION: NCT03996044.

Effects of photobiomodulation in salivary glands of chronic kidney disease patients on hemodialysis.

This randomized placebo-controlled trial evaluates the impact of photobiomodulation (PBMT) on the salivary flow and biochemistry of patients with chronic kidney disease (CKD) on hemodialysis. Forty-four patients on hemodialysis self-responded two questionnaires for oral health and salivary gland function perception. The subjects were evaluated for function of salivary glands and randomly allocated to two groups: PBMT group (three irradiations at 808 nm, 100 mW, 142 J/cm2, and 4 J per site); and placebo group. Patients were submitted to non-stimulated and stimulated sialometry and after the treatment at baseline and 14 days. Salivary volume and biochemical of the saliva were analyzed. At baseline, most subjects had self-perception of poor oral health (52.6%) and salivary dysfunction (63.1%). Clinical exam revealed that 47.3% of subjects presented dry mucosa. PBMT promoted increase of the non-stimulated (p = 0.027) and stimulated saliva (p = 0.014) and decrease of urea levels in both non-stimulated (p = 0.0001) and stimulated saliva (p = 0.0001). No alteration was detected in total proteins and calcium analysis. Patients with kidney disease can present alteration in flow, concentrations, and composition of saliva, affecting oral health, but our findings suggest that PBMT is effective to improve hyposalivation and urea levels in saliva of patients with CKD.

Efficacy of antimicrobial photodynamic therapy (aPDT) for nonsurgical treatment of periodontal disease: a systematic review.

Although the standard treatment for periodontal disease is based on scaling and root planing (SRP), the use of antimicrobial photodynamic therapy (aPDT) has been studied as a complement to obtain better clinical results. The purpose of this study was to evaluate the effect of aPDT as adjuncts to SRP, compared with SRP alone, on clinical parameters of chronic periodontal patients. Only randomized controlled trials with at least 3-month follow-ups, of SRP alone and in association with aPDT, were included. The MEDLINE (PubMed), Google Scholar, and LILACS databases were searched for articles published up to July 2020. Random-effects meta-analyses were conducted for clinical attachment level (CAL) and probing pocket depth (PPD) change after treatment. Of 141 potentially relevant papers, 22 were included. The association between SRP and aPDT promoted a significant CAL gain and PPD reduction. Periodontal treatment was partially improved by aPDT, and a favorable effect of indocyanine green-mediated aPDT was observed, and high concentrations of phenothiazine chloride presented clinical improvement as well.

Photodynamic therapy for endodontic treatment of primary teeth: A randomized controlled clinical trial.

OBJECTIVES: This study aimed to evaluate the reduction in bacterial load following conventional endodontic treatment with and without antimicrobial photodynamic therapy (a-PDT) in primary teeth. METHODS: Thirty primary anterior teeth with a diagnosis of pulp necrosis were selected. Patients were randomly allocated to two groups as follows: Group I, patients undergoing conventional root canal therapy (n = 15) and Group II, patients undergoing conventional root canal therapy combined with antimicrobial PDT (n = 15). For PDT, methylene blue, at a concentration of 0.005 %, was used as the photosensitizing agent, which was applied to the interior of the canal with a sterile paper cone for 3 min, followed by the administration of laser light for 40 s (wavelength: 660 nm, energy density: 4 J/cm², power: 100 mW), delivered in direct contact at the entrance to the root canal. Two microbiological samples of the intra-canal content were taken (one before and one immediately after treatment in both groups) using paper cones. Clinical follow-up involved the investigation of fistulas and mobility and was performed 1- and 3 months after treatment. Data were statistically analyzed. RESULTS: The reduction in bacterial load was 93 % in Group I and 99 % in Group II, with no statistically significant difference. CONCLUSIONS: Conventional treatment combined with antimicrobial PDT with parameters used in this study proved effective but presented equal efficacious capability to conventional endodontic treatment alone. CLINICAL SIGNIFICANCE: The use of PDT has been studied in endodontic treatment protocols in permanent teeth. However, clinical trials in deciduous teeth are necessary for establishing the effectiveness and parameters of this application. The present study analyzes the results of PDT in the endodontic treatment of deciduous teeth.

Antimicrobial photodynamic chemotherapy mediated by PapaMBlue on chronic periodontal disease: Study protocol for a randomized, blind, controlled trial.

BACKGROUND: The elimination of the pathogenic microorganisms of the periodontal pocket is one of the main points for success in periodontal treatment. The objective of this study is to investigate the clinical and antimicrobial effect of papain-mediated photodynamic therapy in the clinical treatment of periodontal disease. METHODS: Twenty patients with chronic periodontitis will be selected. Patients will be randomly divided into 2 groups (n = 10). Group 1 will receive conventional periodontal treatment and group 2 will receive conventional treatment and antimicrobial photodynamic therapy (PACT). Conventional treatment will consist of oral hygiene guidance, with brushing technique instructions and recommendation of daily flossing. The calculus deposits on the teeth will be removed with ultrasound equipment and curettes for scraping and root planning. The PACT will be performed at the end of each periodontal treatment session, at sites with bags ≥4 mm. PapaMblue photosensitizer will be deposited in the periodontal pockets with a syringe and a pre-irradiation time of 1 minute will be adopted. Then, the laser emitting wavelength of 660 nm, with power of 100 mW, for 2 minutes, radiant exposure of 30 J/cm and power density of 250 mW/cm will be applied. Patients will undergo clinical evaluations before treatment (day 1) at 30, 60, and 90 days after the end of treatment; and microbiological evaluations before and immediately after treatment. The distribution of the data within each group and the homogeneity of the variances will be verified. With this information, the most appropriate statistical test in each evaluation will be used. The sample calculation is based on the literature and the significance level of 5% will be adopted. DISCUSSION: The combination of PACT with methylene blue in a papain gel and the conventional treatment may increase the reduction of bacteria in periodontal pockets.

Parameters for antimicrobial photodynamic therapy on periodontal pocket-Randomized clinical trial.

BACKGROUND: Antimicrobial photodynamic therapy (aPDT) has been investigated as an adjunctive to periodontal treatment but the dosimetry parameters adopted have discrepancies and represent a challenge to measure efficacy. There is a need to understand the clinical parameters required to obtain antimicrobial effects by using aPDT in periodontal pockets. The aim of this study was to investigate parameters relating to the antimicrobial effects of photodynamic therapy in periodontal pockets. MATERIAL AND METHODS: This randomized controlled clinical trial included 30 patients with chronic periodontitis. Three incisors from each patient were selected and randomized for the experimental procedures. Microbiological evaluations were performed to quantify microorganisms before and after treatments and spectroscopy was used to identify methylene blue in the pocket. A laser source with emission of radiation at wavelength of ʎ = 660 nm and output radiant power of 100 mW was used for 1, 3 and 5 min. One hundred µM methylene blue was used in aqueous solution and on surfactant vehicle. RESULTS: The results demonstrated the absence of any antimicrobial effect with aqueous methylene blue-mediated PDT. On the other hand, methylene blue in the surfactant vehicle produced microbial reduction in the group irradiated for 5 min (p < 0.05). Spectroscopy showed that surfactant vehicle decreased the dimer peak signal at 610 nm. CONCLUSION: Within the parameters used in this study, PDT mediated by methylene blue in a surfactant vehicle reached significant microbial reduction levels with 5 min of irradiation. The clinical use of PDT may be limited by factors that reduce the antimicrobial effect. Forms of irradiation and stability of the photosensitizers play an important role in clinical aPDT.

The effect of antimicrobial photodynamic therapy on infected dentin in primary teeth: A randomized controlled clinical trial protocol.

BACKGROUND: Antimicrobial photodynamic therapy (aPDT) has been used for the treatment of dental caries. Papacarie is a gel composed of papain and chloramine employed for the partial removal of carious tissue, effective against bacteria, however, some studies report that this antibacterial action is not quite so evident. The aim of this study is to evaluate the clinical effect of aPDT on infected dentin in dental caries lesion in primary teeth. METHODS: Thirty-two primary molars with deep occlusal dental caries will be selected and divided in 2 groups: G1 - caries removal with a low-speed drill and G2 - application of aPDT with PapacarieMBlue. After treatment, all the teeth will be restored with glass ionomer cement and followed up clinically and radiographically, with evaluations at 3, 6, and 12 months. Dentin samples before and after treatment will be analyzed microbiologically. The data will be submitted to descriptive statistical analysis of the association between the categorical variables and both age and gender using the chi-square test and Fisher exact text. The Student t test and analysis of variance will be used for the comparison of mean signs and symptoms of reversible pulpitis. Pearson correlation coefficients will be calculated for the analysis of correlations among the continuous variables. DISCUSSION: Adding methylene blue dye to the formula of PapacarieMBlue might potentiate the antimicrobial action of aPDT and work more effectively on the infected dentin combined with a conservative, minimally invasive treatment. TRIAL REGISTRATION: NCT02734420 on 10 march 2016.

Photodynamic therapy with Bixa orellana extract and LED for the reduction of halitosis: study protocol for a randomized, microbiological and clinical trial.

BACKGROUND: Halitosis is an unpleasant breath odour that can interfere with the professional life, social life and quality of life of people who suffer from it. A modality of treatment that has been increasing in dentistry is antimicrobial photodynamic therapy (aPDT). Bixa orellana, popularly known as "urucum" is a plant native to Brazil. The seeds are used to produce a dye that is largely used in the food, textile, paint and cosmetic industries. The aim of this study is to verify whether aPDT with Bixa orellana extract and blue light-emitting diodes (LEDs) is effective in reducing halitosis. This method will also be compared with tongue scraping, the most commonly used conventional method for tongue coating removal, and the association of both methods will be evaluated. METHODS/DESIGN: A randomized clinical trial will be conducted at the dental clinic of the Universidade Nove de Julho. Thirty-nine patients will be divided by block randomization into three groups (n = 13) according to the treatment to be performed. In Group 1, tongue scraping will be performed by the same operator in all patients for analysis of the immediate results. Patients will also be instructed on how to use the scraper at home. Group 2 will be treated with aPDT with Bixa orellana extract and the LED light curing device: Valo Cordless Ultradent®. Six points in the tongue dorsum with a distance of 1 cm between them will be irradiated. The apparatus will be pre-calibrated at wavelength 395-480 nm for 20 s and 9.6 J per point. In Group 3, patients will be submitted to the tongue scraping procedure, as well as to the previously explained aPDT. Oral air collection with the Oral Chroma™ and microbiological collections of the tongue coating shall be done before, immediately after and 7 days after treatment for comparison. DISCUSSION: Halitosis treatment is a topic that still needs attention. The results of this trial could support decision-making by clinicians regarding aPDT using blue LEDs for treating halitosis on a daily basis, as most dentists already have this light source in their offices. TRIAL REGISTRATION: ClinicalTrials.gov, NCT03346460 . Registered on 17 November 2017.

Controlling methylene blue aggregation: a more efficient alternative to treat Candida albicans infections using photodynamic therapy.

Methylene Blue (MB) has been widely used in antimicrobial Photodynamic Therapy (aPDT), however, the mechanisms of action (Type I or Type II) are defined by its state of aggregation. In this sense, the identification of the relationships between aggregation, the mechanisms of action and the effectiveness against microorganisms, as well as the establishment of the means and the formulations that may favor the most effective mechanisms, are essential. Thus, the objective of this study was to assess the in vitro aPDT efficacies against Candida albicans, by using MB in vehicles which may influence the aggregation and present an oral formulation (OF) containing MB, to be used in clinical aPDT procedures. The efficacy of MB at 20 mg L-1 was tested in a range of vehicles (water, physiological solution - NaCl 0.9%, phosphate saline buffer - PBS, sodium dodecyl sulfate 0.25% - SDS and urea 1 mol L-1) in a C. albicans planktonic culture, when using 4.68 J cm-2 of 640 ± 12 nm LED for the irradiations, as well as 5 minutes of pre-irradiation time, together with measuring the UFC mL-1. Based upon these analyses, an OF containing MB in the most effective vehicle was tested in the biofilms, as a proposal for clinical applications. When comparing some of the vehicles, sodium dodecyl sulfate was the only one that enhanced an MB aPDT efficacy in a planktonic C. albicans culture. This OF was tested in the biofilms and 50 mg L-1 MB was necessary, in order to achieve some reduction in the cell viabilities after the various treatments. The light dosimetries still need further adaptations, in order for this formulation to be used in clinical applications. The present research has indicated that the development of this formulation for the control of MB aggregations may result in more effective clinical protocols.

Influence of Ultrapulsed CO2 Laser, before Application of Different Types of Fluoride, on the Increase of Microhardness of Enamel In Vitro.

OBJECTIVE: To evaluate the influence of ultrapulsed CO2 laser in combination with commercial fluoride products in order to verify the increase of microhardness of artificial enamel caries lesions. MATERIALS AND METHODS: Bovine enamel specimens were prepared, and artificial enamel caries lesions were created. Teeth were randomly divided into 5 groups (n=10): treated with laser (L), laser + neutral fluoride gel 2% (LNF), laser + acidulated phosphate fluoride gel 1.23% (LAFG), laser + acidulated fluoride mousse 1.23% (LAFM), and laser + fluoride varnish 5% (LFV). Microhardness was evaluated at baseline, after caries induction, after CO2 laser irradiation + fluoride treatment in the 1st week, and after fluoride treatment at 3rd and 5th week. RESULTS: There was a decrease in microhardness in all groups after artificial enamel caries lesion formation; no increase in microhardness was found in the first and third weeks in all groups (p > 0.05). In the fifth week, an increase in microhardness occurred in all groups (p < 0.05). CONCLUSION: Although CO2 laser irradiation in combination with different commercial fluoride products was capable of increasing microhardness on enamel caries lesions in bovine tooth enamel it is necessary to confirm these results by testing the isolated effect of fluoride on enamel surface microhardness. Also, although microhardness was higher in the fluoride varnish group than in the other groups in the fifth week it is not possible to discard the best effect of fluoride varnish treatment on absence of artifacts that may occur with the other fluoride treatments. CLINICAL RELEVANCE: In order to prove that CO2 laser may contribute to an increase in microhardness when applied to enamel lesions in combination with different commercial fluoride products it is necessary to conduct additional studies. Also, higher microhardness of fluoride varnish group should be carefully considered.