Arginine and sodium fluoride affect the microbial composition and reduce biofilm metabolism and enamel mineral loss in an oral microcosm model.

OBJECTIVE: To assess the effects of arginine, with or without sodium fluoride (NaF; 1,450 ppm), on saliva-derived microcosm biofilms and enamel demineralization. METHODS: Saliva-derived biofilms were grown on bovine enamel blocks in 0.2 % sucrose-containing modified McBain medium, according to six experimental groups: control (McBain 0.2 %); 2.5 % arginine; 8 % arginine; NaF; 2.5 % arginine with NaF; and 8 % arginine with NaF. After 5 days of growth, biofilm viability was assessed by colony-forming units counting, laser scanning confocal microscopy was used to determine biofilm vitality and extracellular polysaccharide (EPS) production, while biofilm metabolism was evaluated using the resazurin assay and lactic acid quantification. Demineralization was evaluated by measuring pH in the culture medium and calcium release. Data were analyzed by Kruskal-Wallis' and Dunn's tests (p < 0.05). RESULTS: 8 % arginine with NaF showed the strongest reduction in total streptococci and total microorganism counts, with no significant difference compared to arginine without NaF. Neither 2.5 % arginine alone nor NaF alone significantly reduced microbial counts compared to the control, although in combination, a reduction in all microbial groups was observed. Similar trends were found for biofilm vitality and EPS, and calcium released to the growth medium. CONCLUSIONS: 8 % Arginine, with or without NaF, exhibited the strongest antimicrobial activity and reduced enamel calcium loss. Also, NaF enhanced the effects of 2.5 % arginine, yielding similar results to 8 % arginine for most parameters analyzed. CLINICAL SIGNIFICANCE: The results provided further evidence on how arginine, with or without NaF, affects oral microcosm biofilms and enamel mineral loss.

pH-Responsive Chitosan-Doped ZnO Hybrid Hydrogels for the Encapsulation of Bioactive Compounds in Aquaculture.

In this study, we synthesized and characterized pH-responsive Chitosan-AgCl-doped ZnO hybrid hydrogels and evaluated their potential for loading aquaculture bioactive compounds, and assessed their antimicrobial properties against a threatening pathogen associated with disease across a broad spectrum of warm water fish and invertebrates. Hydrogel characterization consisted of assessing morphology via SEM, composition via EDS, hydrogels' network components interactions via FT-IR and pH response through swelling behavior determinations. The swelling characterization of the synthesized hydrogels demonstrated a pH-responsive behavior, showing that low pH values caused the hydrogel polymeric network to expand and capture more of the aqueous solution. These characteristics make the synthesized hydrogels suitable for the encapsulation and controlled release of drugs and bioactive compounds in aquaculture. Chitosan_ZnO hybrid hydrogels showed great antimicrobial activity against Vibrio harveyi, even better than that of loaded PB hydrogels. Here, we provide evidence for the potential capacity of Chitosan_ZnO hybrid hydrogels for the preventive and curative treatment of diseases that impact aquaculture animal health and prevent drug resistance by bacteria.

Cabildo ciudadano sobre eutanasia: participación social en salud y formación ciudadana / Citizen council on euthanasia: social participation in health and citizen formation / Cabildo Cidadão sobre eutanásia: participação social na saúde e formação cidadã

Resumen (analítico) El artículo describe la implementación del cabildo ciudadano sobre eutanasia en niños, niñas y adolescentes en Colombia. Se realizó un estudio cualitativo exploratorio, de tipo investigación-acción-participativa. En la primera fase se conformaron tres paneles: formador, implicados y ciudadanos, y se llevaron a cabo dos etapas de deliberación: formativa y pública. En la fase de análisis de contenido (orientada por preguntas dirigidas a los ciudadanos), se caracterizó al minipúblico involucrado. Se destaca como hallazgos centrales la transformación de la opinión de las y los ciudadanos participantes, así como de sus capacidades deliberativas, lo que permitió dar cuenta de la efectividad de la metodología para facilitar que las y los ciudadanos formulen recomendaciones consensuadas en temas complejos y contribuyan en procesos de formación en democracia y participación social en salud.

Abstract (analytical) The article describes the implementation of the Citizen Council on euthanasia in children and adolescents in Colombia. An exploratory qualitative study using the participatory action research methodology was carried out. In the first phase, three panels were created with trainers, stakeholders and citizens, and two deliberative stages (formative and public) were carried out. In another phase involving content analysis -guided by questions addressed to citizens- a characterization of the minipublic involved in the deliberative exercise was made. The transformation of the opinions of the participating citizens, as well as their deliberative capacities, is a central finding of the study. The authors identified the effectiveness of the methodology to facilitate citizen's design of consensual recommendations about complex issues, as well as contributing to pedagogical processes related to democracy and social participation in health training.

Resumo (analítico) O artigo descreve a implementação do Conselho Cidadão sobre a eutanásia nas crianças e adolescentes na Colômbia. Foi realizado um estudo qualitativo exploratório de pesquisa-ação-participativa. Na primeira fase foram criados três painéis, formadores, intervenientes e cidadãos, e realizadas duas fases de deliberação (formativa e pública). Numa outra fase, de análise de conteúdo -orientada por questões dirigidas aos cidadãos-, foi feita a caracterização do mini público envolvido no exercício deliberativo. A transformação da opinião dos cidadãos participantes, bem como das suas capacidades deliberativas, destaca-se como descoberta central, o que permitiu perceber a eficácia da metodologia para facilitar a formulação de recomendações consensuais sobre questões complexas pelos cidadãos, assim como para contribuir no desenvolvimento de processos pedagógicos de formação em democracia e participação social em saúde.

Assessment of Factors Influencing Personal Exposure to Air Pollution on Main Roads in Bogota: A Mixed-Method Study.

Background and Objectives: Particulate Matter (PM), particles of variable but small diameter can penetrate the respiratory system via inhalation, causing respiratory and/or cardiovascular diseases. This study aims to evaluate the association of environmental particulate matter (PM2.5) and black carbon (BC) with respiratory health in users of different transport modes in four roads in Bogotá. Materials and Methods: this was a mixed-method study (including a cross sectional study and a qualitative description of the air quality perception), in 300 healthy participants, based on an exploratory sequential design. The respiratory effect was measured comparing the changes between pre- and post-spirometry. The PM2.5 and black carbon (BC) concentrations were measured using portable devices. Inhaled doses were also calculated for each participant according to the mode and route. Perception was approached through semi-structured interviews. The analysis included multivariate models and concurrent triangulation. Results: The concentration of matter and black carbon were greater in bus users (median 50.67 µg m-3; interquartile range (-IR): 306.7). We found greater inhaled dosages of air pollutants among bike users (16.41 µg m-3). We did not find changes in the spirometry parameter associated with air pollutants or transport modes. The participants reported a major sensory influence at the visual and olfactory level as perception of bad air quality. Conclusions: We observed greater inhaled doses among active transport users. Nevertheless, no pathological changes were identified in the spirometry parameters. People's perceptions are a preponderant element in the assessment of air quality.

Effects of curcumin-mediated antimicrobial photodynamic therapy associated to different chelators against Enterococcus faecalis biofilms.

BACKGROUND: The aim of this study was to evaluate curcumin-mediated antimicrobial photodynamic therapy (aPDT) action combined or not with ethylenediaminetetraacetic acid (EDTA) and hydroxyethylidene bisphosphonate (HEBP) on Enterococcus faecalis biofilms. METHODS: Enterococcus faecalis biofilms were grown on dentin bovine discs in brain heart infusion (BHI) medium with 1% glucose, in aerobic conditions at 37°C for 7 days. Then, they were randomly distributed to one of experimental conditions, as follows: control, 75 J.cm-2 LED, 600 µmol.L-1 curcumin, 17% EDTA, 18% HEBP, 600 µmol.L-1 curcumin plus 75 J.cm-2 LED, 600 µmol.L-1 curcumin plus 17% EDTA, 600 µmol.L-1 curcumin plus 18% HEBP, 600 µmol.L-1 curcumin plus 17% EDTA and 75 J.cm-2 LED or 600 µmol.L-1 curcumin plus 18% HEBP and 75 J.cm-2 LED. The viability of microorganisms and the vitality of biofilms were determined by colony forming unit counts and confocal scanning laser microscopy (CSLM), respectively. Statistical analysis was conducted by Kruskal Wallis and Dunn's post-hoc tests (α = 0.05). RESULTS: The results showed that all combinations of aPDT with chelators significantly reduced the viability of microbial cells and the vitality of biofilms in comparison to control, even when considering deeper layers of biofilms. CONCLUSION: The combination of curcumin with EDTA and HEBP similarly improved the effect of aPDT on E. faecalis biofilms.

Effect of a sugarcane cystatin on the profile and viability of microcosm biofilm and on dentin demineralization.

To analyze the effect of a sugarcane cystatin (CaneCPI-5) on the microbial profile and viability, as well as on the prevention of dentin demineralization using a microcosm biofilm model. Ninety bovine dentine specimens were divided into five experimental groups according with the solution they were treated for 60 s: (1) PBS (negative control), (2) 0.12% chlorhexidine (positive control), (3) Fluoride (500 ppm F, as NaF), (4) 0.025 mg/ml CaneCPI-5, and (5) 0.05 mg/ml CaneCPI-5. Specimens were incubated with inoculum (McBain's saliva plus human saliva) in the first 8 h, and from then on, they were exposed to McBain saliva containing sucrose and daily treated (60 s) with the solutions for 5 days. Resazurin and colony-forming unit counting assays were performed. Dentin demineralization was measured by transverse micro-radiography (TMR). 0.12% chlorhexidine significantly reduced the metabolic activity of the microcosm biofilm in relation to the negative control and treated groups (p < 0.01). CHX and F significantly reduced the counts of total microorganisms, mutans group streptococci, and lactobacilli when compared with the negative control. None of the treatments was able to significantly reduce dentin demineralization in comparison with the negative control. In the model evaluated, CaneCPI-5 neither altered the microcosm biofilm profile and viability nor protected dentin against demineralization.

In vitro effect of curcumin-mediated antimicrobial photodynamic therapy on fibroblasts: viability and cell signaling for apoptosis.

Although it was demonstrated that curcumin-mediated antimicrobial photodynamic therapy (aPDT) is effective for reducing the viability of microbial cells and the vitality of oral biofilms, the cytotoxicity of this therapeutic approach for host cells has not been yet elucidated. Hence, the aim of this study was to evaluate the cytotoxicity and apoptotic effects of curcumin-mediated aPDT on mouse fibroblasts. Cells were treated with 0.6 or 6 µmol.L-1 curcumin combined with 0.075 or 7.5 J.cm-2 LED at 455 nm. Cytotoxicity was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet (CV) assays, while quantitative reverse transcriptase-PCR (qRT-PCR) was used to assess the expression of Bax, Bad, Bcl-2, VDAC-1, cytochrome C, and Fas-L genes for apoptosis. The differences between groups were detected by Kruskal-Wallis and post hoc Dunn's tests for MTT and CV assays and by ANOVA and post hoc Tukey test for qRT-PCR (P < 0.05). The effect of 0.6 µmol.L-1 curcumin plus 0.075 J.cm-2 LED (minimum parameter) did not differ statistically from control group; however, the combination of 0.6 µmol.L-1 curcumin plus 7.5 J.cm-2 LED reduced viable cells in 34%, while the combinations of 6 µmol.L-1 curcumin plus 0.075 and 7.5 J.cm-2 LED reduced viable cells in 47% and 99%, respectively. aPDT increased significantly the relative expression of Bax/Bcl-2, cytochrome C, VDAC-1, and Fas-L genes, without influence on the ratio Bad/Bcl-2. Therefore, curcumin-mediated aPDT activated Bcl-2 apoptosis signaling pathways in mouse fibroblasts regarding present conditions, reducing the viability of cells with the increase of curcumin concentrations and light energies.

Characterization of polymicrobial biofilms obtained from saliva or carious lesions in dentin.

This study aimed to compare the formation of polymicrobial biofilms using carious dentin or saliva as inoculum for application in in vitro microbiological studies on caries research. For biofilm growth, combined samples of infected dentin or saliva from three donors were used. The biofilms were grown on glass coverslips, under a regimen of intermittent exposure (6 h day-1) to 1% sucrose for 4 days. Total bacterial loads, as well as specific aciduric bacteria and mutans streptococci loads were quantified and correlated with biofilm acidogenicity and susceptibility to chlorhexidine. The data were evaluated using the Student's-t, Mann Whitney and Kruskal-Wallis tests. The two biofilms showed similar microbial loads (total bacteria, aciduric bacteria and mutans streptococci) on day 4, and high acidogenicity after 48 h and were susceptible to chlorhexidine at different time intervals. In conclusion, both dentin and saliva can be used as an inoculum in in vitro studies of processes related to biofilm formation.

Urban Transformations and Health: Methods for TrUST-a Natural Experiment Evaluating the Impacts of a Mass Transit Cable Car in Bogotá, Colombia.

Background: Cable cars provide urban mobility benefits for vulnerable populations. However, no evaluation has assessed cable cars' impact from a health perspective. TransMiCable in Bogotá, Colombia, provides a unique opportunity to (1) assess the effects of its implementation on the environmental and social determinants of health (microenvironment pollution, transport accessibility, physical environment, employment, social capital, and leisure time), physical activity, and health outcomes (health-related quality of life, respiratory diseases, and homicides); and (2) use citizen science methods to identify, prioritize, and communicate the most salient negative and positive features impacting health and quality of life in TransMiCable's area, as well as facilitate a consensus and advocacy-building change process among community members, policymakers, and academic researchers. Methods: TrUST (In Spanish: Transformaciones Urbanas y Salud: el caso de TransMiCable en Bogotá) is a quasi-experimental study using a mixed-methods approach. The intervention group includes adults from Ciudad Bolívar, the area of influence of TransMiCable. The control group includes adults from San Cristóbal, an area of future expansion for TransMiCable. A conceptual framework was developed through group-model building. Outcomes related to environmental and social determinants of health as well as health outcomes are assessed using questionnaires (health outcomes, physical activity, and perceptions), secondary data (crime and respiratory outcomes) use of portable devices (air pollution exposure and accelerometry), mobility tracking apps (for transport trajectories), and direct observation (parks). The Stanford Healthy Neighborhood Discovery Tool is being used to capture residents' perceptions of their physical and social environments as part of the citizen science component of the investigation. Discussion: TrUST is innovative in its use of a mixed-methods, and interdisciplinary research approach, and in its systematic engagement of citizens and policymakers throughout the design and evaluation process. This study will help to understand better how to maximize health benefits and minimize unintended negative consequences of TransMiCable.

Cytotoxic effect and apoptosis pathways activated by methylene blue-mediated photodynamic therapy in fibroblasts.

Antimicrobial photodynamic therapy (aPDT) has been used as an adjuvant treatment of oral infections as a minimal intervention clinical approach. Its antimicrobial efficacy was demonstrated in several studies; however, there is a lack of evidence on its cytotoxic effect on mouse fibroblasts (NIH/3T3). The aim of this study was to evaluate the cytotoxicity and apoptotic pathways of methylene blue-mediated aPDT on mouse fibroblasts. Cells were treated with 0.1 or 1.0 mg.L-1 methylene blue (MB), and 0.075 or 7.5 J.cm-² LED at 630 nm. Cell viability was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet (CV) assays, while cDNA expression for Bax, Bad, Bcl-2, VDAC-1, cytochrome C and Fas-L was assessed by qRT-PCR (1, 3, 6 and 24 h). The differences between groups were detected by Kruskal-Wallis and post-hoc Dunn's tests for MTT and CV assays, and by ANOVA and post-hoc Tukey test for qPCR (P < 0.05). The combination of 1.0 mg.L-1 MB and 7.5 J.cm-² LED significantly reduced the cellular viability, whereas MB and LED alone were innocuous to fibroblasts. MB-mediated aPDT increased the expression of cytochrome C and Fas-L after 3 h, and Bax/Bcl-2, Bad/Bcl-2, and VDAC-1 after 6 h from treatment. Based on these results, MB-mediated aPDT induced cytotoxicity on mouse fibroblasts, with consequent activation of Bcl-2 apoptosis signaling pathways. Further studies are needed to determine the adequate parameters of aPDT to inactivate microorganisms without damaging fibroblasts.

The effectiveness of curcumin-mediated antimicrobial photodynamic therapy depends on pre-irradiation and biofilm growth times.

BACKGROUND: The aim of this study was to determine the influence of distinct pre-irradiation times (PIT) of curcumin on the effectiveness of antimicrobial photodynamic therapy (aPDT) against intact dentin caries biofilms grown for 3 or 5 days. METHODS: The microcosm biofilms grew on non-fluorescent glass blocks immersed in McBain medium with 1% sucrose, using microaerophilic conditions at 37 °C for 3 or 5 days. The biofilms were treated by the association of 600 µmol.L-1 curcumin using different pre-irradiation times (1, 2 or 5 min) combined with 0 or 75 J.cm-2 blue LED. Then, the vitality of biofilms was determined by confocal scanning laser microscopy (CSLM), after being stained with the mixture of ethidium bromide and fluorescein diacetate. Statistical analysis was performed by two-way ANOVA and post-hoc Tukey tests, after arcsine transformation (P < 0,05). RESULTS: In comparison to control, curcumin alone (PIT = 5 min) and all combinations of curcumin and LED reduced significantly the vitality of 3-day biofilms. Distinctly, only curcumin plus LED using PITs of 2 or 5 min were effective in reducing the vitality of 5-day biofilms. CONCLUSION: Curcumin-mediated aPDT significantly decreased the vitality of intact dentin caries microcosms grown during 3 or 5 days, although successful treatments of 5-day biofilms required longer PITs in comparison to their counterparts.

Could a chelant improve the effect of curcumin-mediated photodynamic antimicrobial chemotherapy against dental intact biofilms?

To our knowledge, there is still no evidence in relation to the combination of curcumin with chelants to improve the effects of antimicrobial photodynamic therapy (aPDT) on complex dental caries biofilms. Therefore, the aim of this study was to evaluate the antimicrobial effect of curcumin-ethylenediaminetetraacetic acid (EDTA)-mediated aPDT on the vitality of intact biofilms of dentin caries microcosms. Biofilms were grown on glass slabs in McBain medium plus 1% sucrose in microaerophily at 37 °C for 5 days. Then, biofilms were treated with associations of 600 µmol L-1 curcumin combined or not with 1% EDTA and 37.5 or 75 J cm-2 LED (455 nm). The vitality was determined by a confocal laser scanning microscopy (CLSM) after staining biofilms with a mixture of 2.5 g L-1 fluorescein diacetate and 0.25 g L-1 ethidium bromide. Statistical analysis was conducted by Kruskal-Wallis and post hoc Dunn's test (P < 0.05). Three treatments were able to reduce the vitality of overall biofilms: curcumin + 75 J cm-2 LED, curcumin-EDTA + 37.5 J cm-2 LED, and curcumin-EDTA + 75 J cm-2 LED. Also, the vitality of inner layers of biofilms was significantly reduced only after the combination of aPDT with EDTA. Therefore, the association of curcumin and EDTA improved the antimicrobial effect of aPDT on dentin caries microcosms, considering the application of lower light densities and deeper layers of biofilms.

Could chlorhexidine be an adequate positive control for antimicrobial photodynamic therapy in- in vitro studies?

BACKGROUND: Chlorhexidine digluconate (CHX) is commonly applied as positive control of new antimicrobials, because it is considered the gold-standard for chemical plaque control. The aim of this study was to compare the effect of treatments with curcumin-mediated aPDT and CHX in relation to the viability of specific microorganism groups in two distinct times (immediately and 24 h later). METHODS: Dentin caries microcosms were grown on bovine dentin discs (37 °C, anaerobiosis) for 3 days in the Active Attachment Amsterdam Biofilm Model. The biofilms were treated with 300 µM curcumin and 75 J.cm-² LED, or 0.06% and 0.12% CHX. Then, total microorganisms, total streptococci, mutans streptococci, and total lactobacilli counts were determined. The statistical analysis was conducted by Kruskal-Wallis and post-hoc Dunn's tests (P < 0.05). RESULTS: Curcumin-mediated aPDT (C + L+), 0.06% and 0.12% CHX reduced mutans streptococci counts (0.19, 0.10 and 0.07 log10 respectively) in the immediate analysis. After 24 h, it was observed a re-growth of microorganisms treated by curcumin-mediated aPDT, whereas both CHX concentrations demonstrated a decrease of the viable microorganisms. CONCLUSION: This study confirmed the substantive effect of CHX and the immediate effect of aPDT. The use of a neutralizer solution was important to block the substantivity of CHX and permit its fair comparison with aPDT, allowing its use as a positive control in further studies.

Curcumin-mediated antimicrobial photodynamic therapy reduces the viability and vitality of infected dentin caries microcosms.

BACKGROUND: To our knowledge, there is a lack of evidence on the effect of Antimicrobial Photodynamic Therapy (aPDT) by the application of curcumin against complex biofilms of dental caries lesions. This study aimed to evaluate the viability, vitality, and acid metabolism of infected dentin caries microcosms treated with curcumin-mediated aPDT. METHODS: After microcosm biofilms growing anaerobically on bovine dentin disks immersed in McBain medium with 1% sucrose at 37 °C for 5 days, the biofilms were treated by the association of DMSO water solution or 600 µmol L-1 curcumin with 0, 37.5 or 75 J cm-2 blue LED (455 nm). Then, the colony-forming units (CFU) counts of total microorganisms, total streptococci, mutans streptococci, and total lactobacilli were determined by plating. The lactic acid concentration was analyzed by enzymatic spectrophotometry method, while the vitality of intact biofilms was evaluated by confocal laser scanning microscope (CLSM). Statistical analysis was performed by Kruskal Wallis and post-hoc Dunn's tests (P < 0.05). RESULTS: Curcumin alone did not affect the viability of microorganisms and the vitality of intact biofilms. However, 75 J cm-2 LED alone decreased the total microorganisms and total lactobacilli counts. The combination of curcumin and LED reduced significantly the counts of all microorganism groups and the vitality of intact biofilms. Differences were not observed between the lactic acid concentrations of distinct groups. CONCLUSIONS: Therefore, curcumin-mediated aPDT was effective in reducing the viability and the vitality of infected dentin caries microcosms, without interfering in their acidogenicity.

The Brazilian validation of a health literacy instrument: the newest vital sign.

OBJECTIVE: This study aimed to perform the cross-cultural adaptation and validation of the NVS for Brazilian Portuguese. MATERIAL AND METHODS: Two hundred and fifty adults responded to the adapted version of the NVS, the Brazilian version of the Rapid Estimate of Adult Literacy in Dentistry (BREALD-30), ten questions of the National Functional Literacy Index (NFLI), and a questionnaire about demographic and oral health-related aspects. Statistical analysis determined the internal consistency, stability, difficulty of items, and convergent/discriminant/predictive validities of the NVS. p < .05 was considered significant. RESULTS: The NVS displayed a good internal consistency (Cronbach's alpha = 0.79) and a fair stability (ICC = 0.57; 0.39-0.70 95% CI). Seventy-two percent of participants answered the easier question (#5) correctly, whereas only 28.4% were successful in responding the most difficult question (#1). The convergent validity of the NVS was demonstrated by its significant correlation with the BREALD-30 (Rs = 0.601, p < .001) and NFLI (Rs = 0.544, p < .001). The observation of higher NVS scores among health professionals, white and more educated people confirmed the discriminant validity of the instrument. Moreover, health literacy was a significant predictor of self-reported oral health and reason for dental utilization. CONCLUSION: The NVS demonstrated adequate psychometric properties to be applied in Brazilian oral health epidemiological surveys.

The in vitro effect of Antimicrobial Photodynamic Therapy on dental microcosm biofilms from partially erupted permanent molars: A pilot study.

BACKGROUND: Antimicrobial Photodynamic Therapy (aPDT) could enhance the prevention of dental caries lesions in pits and fissures of partially erupted molars, by killing microorganisms from complex dental biofilms. This pilot study aimed to evaluate the effect of Antimicrobial Photodynamic Therapy (aPDT) on the viability of specific microorganism groups of dental microcosm biofilms from occlusal surfaces of first permanent molars in eruption. METHODS: Dental microcosm biofilms grown on bovine enamel blocks, from dental plaque collected on occlusal surfaces of a partially erupted lower right first permanent molar, with McBain medium plus 1% sucrose in anaerobic condition at 37 °C for 72 h. The experiments were performed in eight groups: L-P- = no treatment (control), L18.75P- = 18.75 J/cm2 LED, L37.5P- = 37.5 J/cm2 LED, L75P- = 75 J/cm2 LED, L-P+ = 200 mM TBO, L18.75P+ = 200 mM TBO + 18.75 J/cm2 LED, L37.5P+ = 200 mM TBO + 37.5 J/cm2 LED, and L75P+ = 200 mM TBO + 75 J/cm2 LED. The counts of total microorganisms, total streptococci and mutans streptococci were determined on selective media agar plates by colony-forming units per mL. The log-transformed counts were analyzed by Kruskal-Wallis and post-hoc Dunn's test (P < 0.05). RESULTS: The counts of all microorganisms treated in the group L75P+ were statistically lower than those treated in L-P-. The aPDT promoted a significant reduction of microorganisms, with a trend of dose-dependent effect. CONCLUSION: TBO-mediated aPDT was effective in reducing the viability of specific microbial groups in dental microcosm biofilms originated from occlusal of permanent molars in eruption.