Analysis of biofilm formation by Candida albicans in different types of orthodontic fixed appliances and devices / Análise da formação de biofilme por Candida albicans em diferentes tipos de aparelhos e dispositivos ortodônticos fixos

Objective: in this study, biofilm formation by Candida albicans in fixed orthodontic appliances was evaluated. Material and Methods: a total of 300 conventional metal brackets (MC), ceramic (CB), self-ligation (SLB), nickel-titanium (NiTi), and nickel-chromium (NiCr) wires, and ligatures types were organized into thirty groups (n=10). To induce biofilm formation, brackets, wires, and ligatures were joined, sterilized, placed in 24-well plates, contaminated with standardized suspensions of C. albicans (107 cells/mL), and incubated at 37 °C for 48 h with shaking. The biofilms formed were detached using an ultrasonic homogenizer, and suspensions were serially diluted and plated on Sabouraud dextrose agar to determine colony-forming units per mL. Scanning electron microscopy was performed before and after the biofilm formation. Results: lower amount of biofilm formation was observed in the MC group than in the CB and SLB groups (p<0.0001). SLB and CB showed similar biofilm formation rates (p=0.855). In general, the cross-sectional wires .018"x.025" showed higher biofilm formation when associated with the three types of brackets. When brackets, wires, and ligatures were associated, the sets with NiCr wires and SSL ligatures with MC brackets (p=0.0008) and CB (p=0.0003) showed higher biofilm formation. Conclusion: thus, brackets of MC with NiTi and NiCr wires showed lower biofilm formation, regardless of the ligature and cross-sectional or gauge of the wire and, MC and CB brackets with NiCr wires and SSL ligatures were more likely to accumulate biofilms (AU)

Objetivo: neste estudo, a formação de biofilme por Candida albicans em aparelhos ortodônticos fixos foi avaliada. Material e Métodos: um total de 300 bráquetes metálicos convencionais (MC), cerâmicos (CB), autoligados (SLB), com fios de níquel-titânio (NiTi) e níquel-cromo (NiCr) e tipos de ligaduras foram organizados em trinta grupos (n=10). Bráquetes, fios e ligaduras foram unidos, esterilizados, colocados em placas de 24 poços, contaminados com suspensões padronizadas de C. albicans (107 células/mL) e incubados a 37°C por 48 h para a formação de biofilmes. Os biofilmes formados foram rompidos por meio de um homogeneizador ultrassônico e suspensões foram diluídas e semeadas em ágar Sabouraud-dextrose para determinar as unidades formadoras de colônias por mL. A microscopia eletrônica de varredura foi realizada antes e após a formação do biofilme. Resultados: foi observada menor formação de biofilme no grupo MC em comparação aos grupos CB e SLB (p<0,0001). A formação de biofilme foi semelhante nos grupos SLB e CB (p=0,855). Em geral, os fios de seção transversal .018"x.025" apresentaram maior formação de biofilme quando associados aos três tipos de bráquetes. Os conjuntos com fios de NiCr e ligaduras SSL com bráquetes MC (p=0,0008) e CB (p=0,0003) apresentaram maior formação de biofilme. Conclusão: bráquetes MC com fios de NiTi e NiCr apresentaram menor formação de biofilme, independente da ligadura e secção transversal ou bitola do fio e, braquetes MC e CB com fios de NiCr e ligaduras SSL foram mais propensos a acumular biofilmes.(AU)

Effects of Bacillus subtilis on Candida albicans: biofilm formation, filamentation and gene expression / Efeitos de Bacillus subtilis sobre Candida albicans: formação de biofilme, filamentação e expressão gênica

Objective: The aim of this study was evaluate the effect of Bacillus subtilis on Candida albicans biofilm formation and filamentation by evaluating the gene expression of ALS3, HWP1, BCR1, EFG1 and TEC1. Material and Methods: Mixed (C. albicans / B.subtilis) and monotypic biofilms were cultured in plates at 37°C for 48 h under shaking for counting viable cells (CFU / mL) and analysis of gene expression by real-time PCR. The C. albicans filamentation assay was performed in medium containing 10% fetal bovine serum at 37°C for 6 hours. Data was analysed by t-Student and Mann­ Whitney tests. Results: B. subtilis reduced the biofilm formation of C. albicans in 1 log when cultured in the same environment (p<0.0001). In addition, it significantly reduced the yeast - hypha transition affecting the morphology of C. albicans. Among all of the analyzed genes, the ALS3 and HWP1 genes were the most affected, achieving 111.1- and 333.3- fold decreases in the C. albicans biofilms associated with B. subtilis, respectively. Conclusion: B. subtilis reduced the biofilm formation and filamentation of C. albicans by negatively regulating the ALS3, HWP1, BCR1, EFG1 and TEC1 genes that are essential for the production of biofilm and hyphae. (AU)

Objetivo: O objetivo deste estudo foi avaliar o efeito de Bacillus subtilis sobre a formação de biofilme e filamentação de Candida albicans através da avaliação da expressão dos genes ALS3, HWP1, BCR1, EFG1 and TEC1. Material e métodos: Biofilmes monotípicos e mistos (C. albicans / B.subtilis) foram cultivados em placas a 37°C por 48 h sob agitação, para a contagem de células viáveis (UFC/mL) e para a análise da expressão gênica por PCR em tempo real. O ensaio de filamentação de C. albicans foi realizado em meio contendo 10% de soro fetal bovino a 37°C por 6 h. Os dados obtidos foram analisados por testes t-Student e Mann­Whitney. Resultados: B.subtilis reduziu em 1 log a formação de biofilme por C. albicans quando cultivados no mesmo ambiente (p<0.0001). Além disso, reduziu significantemente a transição de levedura para hifa, afetando assim, a morfologia de C. albicans. Em relação aos genes analisados, os genes ALS3 e HWP1 foram os mais regulados negativamente, com uma diminuição de 111,1 e 333,3 vezes, respectivamente, na sua expressão em biofilmes de C. albicans associados a B. subtilis. Conclusão: B. subtilis reduziu a filamentação e a formação de biofilme de C. albicans através da regulação negativa dos genes ALS3, HWP1, BCR1, EFG1 e TEC1, que são essenciais na produção de hifas e de biofilme. (AU)

Assessment of Candida SPP. proliferation in occlusal and palatal splints / Avaliação da proliferação Candida SPP. em placas oclusais e palatais

Candida species inhabit the oral cavity of all individuals who wear complete denture and whose material is the same as that used in splints. Assess the growth of C. albicans in occlusal and palatal splints used for treatment of TMD so that the potential risks of oral microbiota can be assessed. The growth of Candida spp. was assessed in the saliva of 27 individuals wearing splints for treatment of TMD. They were divided into two groups: G1 (n = 14), individuals wearing occlusal splint; and G2 (n = 13), individuals wearing palatal splint. Saliva samples were collected during placement of the splints (T1) and after 4 months (T2), being stored in PBS (10 mL) after 60-second rinses. It was observed that patients wearing occlusal splints (G1) had an increase of 0.648 CFU/mL (Log 10), with statistically significant differences (P = 0.043) for C. albicans (42.33%), C. glabrata (5.52%), C. krusei (41.72%) and C. tropicalis (10.43%). In the group of patients wearing palatal splints (G2), there was a decrease of 0.101 CFU/mL (Log 10), was observed with (P = 0.964) only the presence of C. albicans. The results suggest that growth of Candida species was greater in patients wearing occlusal splints compared to those wearing palatal ones as the presence of different yeast species was found in the former.

Espécies de Candida habitam a cavidade oral de 60-100% de indivíduos usuários de prótese total, cujo material é o mesmo utilizado em placas miorrelaxante. Avaliar o crescimento de C. albicans. em placas relaxantes musculares oclusais e palatais, usadas para o tratamento de DTM, na intenção de verificar riscos em potencial à microbiota bucal. Avaliou-se o crescimento de Candida spp. na saliva de 27 indivíduos, usuários de placa miorrelaxante, em tratamento para DTM no ICT-UNESP. Os indivíduos foram divididos em dois grupos: G1(n=14) ­ placa com recobrimento oclusal; e G2 (n=13) ­ sem recobrimento. As coletas foram com PBS (10mL), em bochechos por 60seg, na instalação das placas (T1) e após 4 meses (T2). Observou-se que pacientes usuários da placa miorrelaxante com recobrimento oclusal (grupo G1) apresentaram aumento de 0,648 UFC/mL (Log10) com diferença estatisticamente significante (p=0,043) analisando-se 42,33% C. albicans, 5,52% C. glabrata, 41,72% C. krusei e 10,43% C. tropicalis. No grupo de pacientes que utilizaram a placa sem recobrimento (grupo G2), observou-se diminuição de 0,101 UFC/mL (Log10) com (p=0,954) apresentando apenas C. albicans. Os resultados sugerem que os pacientes que fizeram uso de placa miorrelaxante com recobrimento oclusal apresentaram maior crescimento de Candida spp. em relação aos usuários de placa sem recobrimento, verificando-se a presença de diferentes espécies da levedura.

Study of Microbial Interaction Formed by "Candida krusei" and "Candida glabrata": "In Vitro" and "In Vivo" Studies

Abstract Recently, the non-albicans Candida species have become recognized as an important source of infection and oral colonization by association of different species in a large number of immunosuppressed patients. The objective of this study was to evaluate the interactions between C. krusei and C. glabrata in biofilms formed in vitro and their ability to colonize the oral cavity of mouse model. Monospecies and mixed biofilms were developed of each strain, on 96-well microtiter plates for 48 h. These biofilms were analyzed by counting colony-forming units (CFU/mL) and by determining cell viability, using the XTT hydroxide colorimetric assay. For the in vivo study, twenty-four mice received topical applications of monospecie or mixed suspensions of each strain. After 48 h, yeasts were recovered from the mice and quantified by CFU/mL count. In the biofilm assays, the results for the CFU/mL count and the XTT assay showed that the two species studied were capable of forming high levels of in vitro monospecie biofilm. In mixed biofilm, the CFU of C. krusei increased (p=0.0001) and C. glabrata decreased (p=0.0001). The metabolic activity observed in XTT assay of mixed biofilm was significantly reduced compared with a single C. glabrata biofilm (p=0.0001). Agreeing with CFU in vitro count, C. glabrata CFU/mL values recovered from oral cavity of mice were statistically higher in the group with single infection (p=0.0001) than the group with mixed infection. We concluded that C. krusei inhibits C. glabrata and takes advantage to colonize the oral cavity and to form biofilms.

Resumo Recentemente, as espécies não albicans tem se tornado uma importante fonte de infecção e de colonização oral pela associação de espécies em um grande número de pacientes imunossuprimidos. O objetivo desse estudo foi avaliar a interação entre C. krusei e C. glabrata em biofilmes formados in vitro e sua capacidade em colonizar a cavidade oral em modelo de camundongo. Biofilmes monoespécies e mistos foram formados em placas de 96 poços por 48 h. Esses biofilmes foram analisados pela contagem de UFC/mL e pela determinação da viabilidade celular, usando ensaio de XTT. Para o estudo in vivo, vinte e quatro camundongos receberam aplicações tópicas de suspensões monoespécies e mistas de cada espécie. Após 48 h, as leveduras foram recuperadas dos camundongos e quantificadas por UFC/mL. Nos ensaios de biofilme, os resultados da contagem de UFC/mL e do ensaio de XTT mostraram que as duas espécies estudadas foram capazes de formar grande quantidade de biofilme monoespécie in vitro. Nos biofilmes mistos, a UFC/mL de C. krusei aumentou (p=0,0001) e de C. glabrata diminuiu (p=0,0001). A atividade metabólica observada no ensaio de XTT nos biofilmes mistos foi significantemente reduzida comparada com o biofilme formado apenas de C. glabrata (p=0,0001). Concordado com as contagens in vitro, os valores de UFC/mL de C. glabrata recuperados da cavidade oral dos camundongos foram estatisticamente maior no grupo com infecção simples (p=0,0001) do que do grupo com infecção mista. Nós concluímos que C. krusei inibe C. glabrata e possui vantagem em colonizar a cavidade oral e formar biofilmes.

Antibiofilm activity in vitro of Rosmarinus officinalis and Syzygium cumini glycolic extracts on Staphylococcus spp. of dentistry interest / Atividade antibiofilme in vitro dos extratos glicólicos de Rosmarinus officinalis e Syzygium cumini em Staphylococcus spp. de interesse odontológico

Objectives: The aim of this study was to identify the slime production and evaluate the effects of Rosmarinus officinalis (rosemary) and Syzygium cumini (jambolan) glycolic extracts, and 0.12% chlorhexidine (CHX) in biofilms formed by strains of coagulase-positive Staphylococcus - CPS and coagulase negative Staphylococcus - CNS isolated from the oral cavity. Material and Methods: Slime production was evaluated by two methods: the color of colony presented in Congo red agar, and through the amount of slime adhered to polystyrene. Biofilms were grown in acrylic resin discs immersed in broth, inoculated with microbial suspension (106 cells/ml) and incubated at 37°C/48 h. After formation, the biofilms were exposed for 5 minutes to glycol extracts, CHX or saline solution. The viability of biofilms was determined by counting the colony-forming units per milliliter (CFU/ml) in agar, and analyzed statistically by Tukey test (p< 0.05). Results: The strains S. aureus, S. schleiferi and S. epidermidis obtained the highest values of slime adhered to polystyrene. R. officinalis promoted reductions ranging from 12.1% to 78.7% in biofilms formed by isolates of CPS, and 9.2% to 73.7% in the biofilms of CNS. S. cumini reduced 12% to 55.7% in biofilms of CPS, and 7.9% to 71.5% in biofilms of CNS. With exception of S. saprophyticus, glycol extracts produced significant reductions in biofilms. For five isolates studied, R. officinalis produced greater reductions than CHX. Conclusion: R. officinalis and S. cumini showed effective antibiofilm activity against isolates that showed slime production.(AU)

Objetivos: O objetivo deste estudo foi identificar a produção de slime e avaliar os efeitos dos extratos glicólicos de Rosmarinus officinalis (alecrim), Syzygium cumini (jambolão) e 0,12% de clorexidina (CLX) em biofilmes formados por cepas de Staphylococcus coagulase positivo (SCP) e Staphylococcus coagulase negativo (SCN) da cavidade oral. Material e Métodos: A produção de slime foi avaliada por dois métodos: a cor da colônia apresentada em ágar vermelho Congo e pela quantidade de slime aderido ao poliestireno. Os biofilmes foram crescidos em discos de resina acrílica imersos em caldo, inoculados com suspensão microbiana (106 células/ml) e incubados a 37°C/48h. Após a formação, os biofilmes foram expostos durante 5 minutos aos extractos glicólicos, CLX ou solução salina. A viabilidade dos biofilmes foi determinada pela contagem das unidades formadoras de colônias por mililitro (UFC/ml) em ágar e analisada estatisticamente pelo teste de Tukey (p< 0,05). Resultados: As cepas S. aureus, S. schleiferi e S. epidermidis obtiveram os maiores valores de aderência ao poliestireno. R. officinalis promoveu reduções variando de 12,1% a 78,7% em biofilmes formados por isolados de SCP e 9,2% a 73,7% nos biofilmes de SCN. S. cumini reduziu de 12% a 55,7% nos biofilmes de SCP, e 7,9% a 71,5% nos biofilmes de SCN. Com exceção de S. saprophyticus, os extratos glicólicos produziram reduções estatísticas nos biofilmes. Para cinco isolados estudados, R. officinalis produziu maiores reduções do que CLX. Conclusão: R. officinalis e S. cumini mostraram atividade antibiofilme efetiva contra isolados que apresentaram produção de slime.(AU)

Mixed biofilms formed by C. albicans and non-albicans species: a study of microbial interactions

Abstract Most Candida infections are related to microbial biofilms often formed by the association of different species. The objective of this study was to evaluate the interactions between Candida albicans and non-albicans species in biofilms formed in vitro. The non-albicans species studied were:Candida tropicalis, Candida glabrata andCandida krusei. Single and mixed biofilms (formed by clinical isolates of C. albicans and non-albicans species) were developed from standardized suspensions of each strain (107 cells/mL), on flat-bottom 96-well microtiter plates for 48 hour. These biofilms were analyzed by counting colony-forming units (CFU/mL) in Candida HiChrome agar and by determining cell viability, using the XTT 2,3-bis (2-methoxy-4-nitro-5-sulphophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide colorimetric assay. The results for both the CFU/mL count and the XTT colorimetric assay showed that all the species studied were capable of forming high levels of in vitro biofilm. The number of CFU/mL and the metabolic activity of C. albicans were reduced in mixed biofilms with non-albicans species, as compared with a singleC. albicans biofilm. Among the species tested, C. krusei exerted the highest inhibitory action against C. albicans. In conclusion, C. albicans established antagonistic interactions with non-albicans Candida species in mixed biofilms.

Efeitos da terapia fotodinâmica antimicrobiana em leveduras do gênero Candida / Effects of antimicrobial photodynamic therapy on Candida yeasts

O objetivo deste estudo foi avaliar os efeitos da terapia fotodinâmica utilizando azul de metileno em cepas de Candida, Foram estudados: 5 C, albicans, 4 C, tropicalis , 4 C, glabrata, 2 C, parapsilosis, 2 C, kefyr, 1 C, stellatoidea, 1 C, krusei e 1 C, lipolytica, Cada cepa foi submetida a quatro condições experimentais: laser e azul de metileno (L+F+), irradiação com laser (L+F-), tratamento com azul de metileno (L-F+) e tratamento com soro fisiológico como grupo de controle (L-F-), Após o tratamento de cada cepa diluições seriadas e plaqueamento em Sabouraud dextrose agar foram realizadas, Os dados de unidades formadoras de colônias por mililitro (CFU/ml) foram analisados pelos testes de ANOVA e Tukey (p <0,05), Os resultados sugerem que os grupos tratados com laser (L+F+ e L+F-) apresentaram médias de UFC/ml (Log) inferiores aos grupos tratados sem laser (L-F- e L-F+), O grupo com a terapia fotodinâmica (L+F+) apresentou uma média de CFU/ml (Log) semelhante ao grupo (L+F-), Concluiu-se que as cepas de Candida analisadas foram sensíveis à irradiação do laser de baixa potência na presença ou ausencia do azul de metileno...

The aim of this study was to evaluate the effects of photodynamic therapy using methylene blue on Candida strains. Were studied: 5 C. albicans, 4 C. tropicalis, 4 C. glabrata 2 C. parapsilosis, 2 C. kefyr, 1 C. stellatoidea, 1 C. krusei and 1 C. lipolytica. Each strain was underwent to four experimental conditions: methylene blue laser (L+P+) irradiation with laser (L+P-), treatment with methylene blue (L-P+), and treatment with saline as control group (L-P-). After treatment of each strain and plating serial dilutions on Sabouraud dextrose agar were performed. The data from colony forming units per milliliter (CFU/ml) were analyzed by ANOVA and Tukey test (p <0.05). The results suggest that the groups treated with laser (L+ P+ and L+P-) showed average CFU/ml (Log) lower than without laser treatment groups (L-F- e L-F+). The group with photodynamic therapy (L+P+) showed an average CFU/ml (Log) similar to the group (L+P-). It was concluded that the strains of Candida analyzed were sensitive to laser irradiation of low power in the presence or absence of methylene blue...

Morphological and chemical changes of dentin after applying different sterilization methods / Avaliação das alterações morfológicas e químicas da dentina após a aplicação de diferentes métodos de esterilização

Aim: The present study evaluated the morphological and chemical changes of dentin produced by different sterilization methods, using scanning electron microscopy (SEM) and energy-dispersive X-ray spectrometry (EDS) analysis. Material and method: Five human teeth were sectioned into 4 samples, each divided into 3 specimens. The specimens were separated into sterilization groups, as follows: wet heat under pressure; cobalt 60 gamma radiation; and control (without sterilization). After sterilization, the 60 specimens were analyzed by SEM under 3 magnifications: 1500X, 5000X, and 10000X. The images were analyzed by 3 calibrated examiners, who assigned scores according to the changes observed in the dentinal tubules: 0 = no morphological change; 1, 2 and 3 = slight, medium and complete obliteration of the dentinal tubules. The chemical composition of dentin was assessed by EDS, with 15 kV incidence and 1  µm penetration. Result: The data obtained were submitted to the statistical tests of Kruskall-Wallis and ANOVA. It was observed that both sterilization methods – with autoclave and with cobalt 60 gamma radiation - produced no significant changes to the morphology of the dentinal tubules or to the chemical composition of dentin. Conclusion: Both methods may thus be used to sterilize teeth for research conducted in vitro. .

Objetivo: O presente estudo teve como objetivo avaliar as alterações morfológicas e químicas na dentina geradas por diferentes métodos de esterilização, através de microscopia eletrônica de varredura (MEV) e da análise por espectrometria de energia dispersiva de raios-X. Material e método: Seccionaram-se 5 dentes humanos em 4 amostras, as quais foram divididas em 3 espécimes cada, totalizando 60 espécimes. Os espécimes foram separados conforme os grupos de esterilização: calor úmido sob pressão; radiação gama cobalto 60; controle (sem esterilização). Após a esterilização, os 60 espécimes foram analisados por MEV, usando 3 aumentos: 1500X, 5000X, 10000X. As imagens foram analisadas por 3 examinadores previamente calibrados, que distribuíram escores referentes às alterações observadas nos túbulos dentinários: 0 = sem alteração morfológica, 1 = baixa obliteração dos túbulos dentinários, 2 = moderada obliteração, 3 = completa obliteração. A composição química da dentina foi avaliada por meio do processo EDS, com 15 kV de incidência e penetração de 1 µm. Os dados obtidos foram submetidos ao teste estatístico de Kruskall-Wallis com nível de significância de 5%. Resultado: De acordo com os resultados obtidos, verificou-se que a esterilização tanto por autoclave quanto por radiação gama cobalto 60 não provocou alterações significativas na morfologia dos túbulos dentinários e na constituição química da dentina. Conclusão: Concluiu-se que ambos os métodos podem ser utilizados para a esterilização de dentes em pesquisa in vitro. .

Different Extracts of Zingiber officinale Decrease Enterococcus faecalis Infection in Galleria mellonella

Dried, fresh and glycolic extracts of Zingiber officinale were obtained to evaluate the action against G. mellonella survival assay against Enterococcus faecalis infection. Eighty larvae were divided into: 1) E. faecalis suspension (control); 2) E. faecalis + fresh extract of Z. officinale (FEO); 3) E. faecalis + dried extract of Z. officinale (DEO); 4) E. faecalis + glycolic extract of Z. officinale (GEO); 5) Phosphate buffered saline (PBS). For control group, a 5 μL inoculum of standardized suspension (107 cells/mL) of E. faecalis (ATCC 29212) was injected into the last left proleg of each larva. For the treatment groups, after E. faecalis inoculation, the extracts were also injected, but into the last right proleg. The larvae were stored at 37 °C and the number of dead larvae was recorded daily for 168 h (7 days) to analyze the survival curve. The larvae were considered dead when they did not show any movement after touching. E. faecalis infection led to the death of 85% of the larvae after 168 h. Notwithstanding, in treatment groups with association of extracts, there was an increase in the survival rates of 50% (GEO), 61% (FEO) and 66% (DEO) of the larvae. In all treatment groups, the larvae exhibited a survival increase with statistically significant difference in relation to control group (p=0.0029). There were no statistically significant differences among treatment groups with different extracts (p=0.3859). It may be concluded that the tested extracts showed antimicrobial activity against E. faecalis infection by increasing the survival of Galleria mellonella larvae.

Extratos seco, fresco e glicólico de Zingiber officinale foram obtidos para avaliar suas ações por meio de ensaio de sobrevivência em G. mellonella contra infecção por Enterococcus faecalis. Oitenta larvas foram divididas em: 1) Suspensão de E. faecalis (controle); 2) E. faecalis + extrato fresco de Z. officinale (FEO); 3) E. faecalis + extrato seco de Z. officinale (DEO); 4) E. faecalis + extrato glicólico de Z. officinale (GEO); 5) Solução tampão fosfato salina (PBS). Para o grupo de controle, 5 µL de inóculo de suspensão padronizada (107 células/mL) de E. faecalis (ATCC 29212) foi injetado na última proleg esquerda de cada lagarta. Para os grupos com tratamento, após a injeção de E. faecalis, os extratos foram injetados na última proleg direita. Após as injeções, as lagartas foram armazenadas a 37 °C e o número de animais mortos foi registrado diariamente em 168 h (7 dias) para analisar a curva de sobrevivência. As lagartas foram consideradas mortas quando elas não mostraram qualquer movimento após o toque. A infecção por E. faecalis levou à morte de 85% das lagartas após 168 h. Não obstante, nos grupos de tratamento com associação dos extratos, houve um aumento nas taxas de sobrevivência de 50% (GEO), 61% (FEO) e 66% (DEO) das lagartas. Em todos os grupos com tratamento, as lagartas apresentaram um aumento na sobrevivência, com diferença estatisticamente significativa em relação ao grupo controle (p=0,0029). Não houve diferença estatisticamente significativa entre os tratamentos com os diferentes extratos (p=0,3859). Pode concluir-se que os extratos testados mostraram atividade antimicrobiana contra a infecção por E. faecalis, aumentando a sobrevivência das lagartas de G. mellonella.

Candida spp. adherence to oral epithelial cells and levels of IgA in children with orthodontic appliances

Adhesion and colonization of the oral cavity by Candida albicans is an initial step in candidosis. Orthodontic and other oral appliances seem to favor candidal presence. The aim of this work was to compare the presence of Candida species in saliva, their adherence to oral epithelial cells, and the levels of anti-C. albicans IgA in children with or without orthodontic appliances. This study included 30 children 5 to 12 years old (9.1 ± 1.7 years old) who were users of removable orthodontic devices for at least 6 months and 30 control children of similar ages (7.7 ± 1.5 years old). The presence of yeast species in the saliva was evaluated by microbiological methods. Candida species were identified using phenotypic methods. Anti-C. albicans IgA levels in saliva were analyzed by ELISA. The yeasts adhering to oral epithelial cells were assessed by exfoliative cytology. No statistically significant differences were observed for saliva yeast counts and anti-C. albicans IgA levels between the studied groups. Children with orthodontic devices exhibited more yeast cells adhering to oral epithelial cells and a higher percentage of non-albicans species relative to the control group. In conclusion, orthodontic appliances may favor the adherence of Candida to epithelial cells but do not influence the presence of these yeasts in saliva, and the levels of anti-C. albicans IgA do not correlate with yeast adherence or presence of Candida in the oral cavity.

The antimicrobial effects of Citrus limonum and Citrus aurantium essential oils on multi-species biofilms

The aim of this study was to evaluate the effects of Citrus limonum and Citrus aurantium essential oils (EOs) compared to 0.2% chlorhexidine (CHX) and 1% sodium hypochlorite (NaOCl) on multi-species biofilms formed by Candida albicans, Enterococcus faecalis and Escherichia coli. The biofilms were grown in acrylic disks immersed in broth, inoculated with microbial suspension (106 cells/mL) and incubated at 37°C / 48 h. After the biofilms were formed, they were exposed for 5 minutes to the solutions (n = 10): C. aurantium EO, C. limonum EO, 0.2% CHX, 1% NaOCl or sterile saline solution [0.9% sodium chloride (NaCl)]. Next, the discs were placed in sterile 0.9% NaCl and sonicated to disperse the biofilms. Tenfold serial dilutions were performed and the aliquots were seeded onto selective agar and incubated at 37°C / 48 h. Next, the number of colony-forming units per milliliter was counted and analyzed statistically (Tukey test, p ≤ 0.05). C. aurantium EO and NaOCl inhibited the growth of all microorganisms in multi-species biofilms. C. limonum EO promoted a 100% reduction of C. albicans and E. coli, and 49.3% of E. faecalis. CHX was less effective against C. albicans and E. coli, yielding a reduction of 68.8% and 86.7%, respectively. However, the reduction of E. faecalis using CHX (81.7%) was greater than that obtained using C. limonum EO. Both Citrus limonum and Citrus aurantium EOs are effective in controlling multi-species biofilms; the microbial reductions achieved by EOs were not only similar to those of NaOCl, but even higher than those achieved by CHX, in some cases.

Comparison of disinfection protocol of irreversible hydrocolloid (alginate) impressions through plastic and metallic trays / Comparação do protocolo de desinfecção de moldes com hidrocolóide irreversível (alginato) em moldeiras plásticas e metálicas

Introduction: Many studies have evaluated the disinfection of irreversible hydrocolloid impressions through different disinfecting agents. However, impression trays can be source of cross-infection requiring disinfection. This study aimed to determine which would be the most suitable tray (metallic or plastic), available in dental market, and the ideal time to achieve disinfection by using 1% sodium hypochlorite poured into the alginate impression. Material and method: Thirty dental impressions from the patients aged from 7-12 years and treated in the Discipline of Orthodontics of the institution were divided into two groups according to the impression tray type: 15 impressions through plastic tray (Morelli) and 15 impressions through metallic tray (Tecnodent). The material collection was performed before and after the application of 1% sodium hypochlorite for 3, 5 and 10 min. After the incubation period of 48 h at 37 ºC, the microorganism colonies were counted on the plates presenting from 30 to 300 colonies to determine the colony-forming unit (CFU) per mL. CFU/ mL results were transformed into logarithm and submitted to statistical analysis by applying ANOVA and Tukey test (p ≤ 0.05). Results: Greater CFU percentage reduction occurred in alginate after three min, in both tray types. Concerning to tray types, it could be observed that the plastic tray showed 100% of reduction after 5 min while the metallic tray exhibited 81.49% of reduction after 3 min. Conclusion: 1 – The plastic tray showed the most effective disinfection after 5 min, with 100% of CFU reduction; 2 – The most effective time of disinfection with 1% sodium hypochlorite poured into the impression was 5 min, for both tray types.

Introdução: Muitos são os trabalhos que avaliam a desinfecção das moldagens com hidrocolóide irreversível e com diferentes agentes desinfetantes, mas as moldeiras também podem ser vetores de infecção cruzada e é necessária a sua desinfecção. A proposta do trabalho foi determinar qual a melhor moldeira (metálica ou plástica), disponíveis no mercado para a utilização do cirurgião dentista, e o tempo ideal para se obter a desinfecção utilizando o hipoclorito de sódio a 1% vertido na moldagem de com alginato. Material e método: Foram obtidas 30 moldagens de pacientes em tratamento na Disciplina de Ortodontia, do Curso de Odontologia, do ICT-UNESP-SJC, com idades entre 7 e 12 anos, divididas em dois grupos de acordo com o tipo de moldeira empregada: 15 moldagens com moldeiras de plástico (Morelli) e 15 com moldeiras de metal (Tecnodent). A coleta de material foi realizada antes e após a aplicação do hipoclorito de sódio a 1% durante 3, 5 e 10 min. Após período de incubação de 48 h a 37 ºC foram contadas colônias de microorganismos nas placas que apresentaram de 30 a 300 colônias para determinação de Unidades Formadoras de Colônias (UFC) por mL. Os resultados em UFC/ mL foram transformados em logaritmo e submetidos à análise estatística Anova e teste de Tukey (p ≤ 0,05). Resultados: No alginato ocorreu uma maior redução percentual de UFC após 3 min, em ambas as moldeiras. Em relação a maior redução nas moldeiras, pudemos observar que a moldeira plástica ocorreu 100% de redução após 5 min e 81,49% de redução na moldeira metálica após 3 min. Conclusão: 1 - A moldeira plástica apresentou desinfecção mais eficiente, após 5 min, com redução de 100% de UFC; 2 - O tempo mais eficaz de desinfecção da moldagem com hipoclorito de sódio a 1% vertido na moldagem foi de 5 min, para moldeira de plástico e de metal.

Correlation of phospholipase and proteinase production of Candida with in vivo pathogenicity in Galleria mellonella

An essential factor to the virulence of the genus Candida is the ability to produce enzymes and this may be crucial in the establishment of fungal infections. AIM:This study investigated in vitro enzymatic activities of Candida species and their virulence in an in vivo Galleria mellonella experimental model. METHODS: Twenty-four clinical strains of Candida spp. isolated from the human oral cavity were evaluated, including the following species: C. albicans, C. dubliniensis, C. glabrata, C. tropicalis, C. krusei, C. parapsilosis, C. norvegensis, C. lusitaniae and C. guilliermondii. All Candida strains were tested in vitro for production of proteinase and phospholipase. The Candida strains were also injected into Galleria mellonella larvae to induce experimental candidiasis, and after 24 hours, the survival rate was assessed. RESULTS: Phospholipase and proteinase activity were observed in 100% of the C. albicans strains. In the non-albicans species, proteinase and phospholipase activity were observed in 25 and 43% of the studied strains, respectively. The most pathogenic Candida species in G. mellonella were C. albicans, C. dubliniensis and C. lusitaniae, whereas C. glabrata was the least virulent species. Furthermore, a positive significant correlation was found between both enzymatic activities with virulence in G. mellonella. CONCLUSIONS: The virulence of Candida strains in G. mellonella is related to the quantity of proteinases and phospholipases production of each strain.

In vitro antimicrobial activity of auxiliary chemical substances and natural extracts on Candida albicans and Enterococcus faecalis in root canals

ABSTRACT Objective: The aim of this study was to evaluate the antimicrobial activity of auxiliary chemical substances and natural extracts on Candida albicans and Enterococcus faecalis inoculated in root canals. Material and Methods: Seventy-two human tooth roots were contaminated with C. albicans and E. faecalis for 21 days. The groups were divided according to the auxiliary chemical substance into: G1) 2.5% sodium hypochlorite (NaOCl), G2) 2% chlorhexidine gel (CHX), G3) castor oil, G4) glycolic Aloe vera extract, G5) glycolic ginger extract, and G6) sterile saline (control). The samples of the root canal were collected at different intervals: confirmation collection, at 21 days after contamination; 1st collection, after instrumentation; and 2nd collection, seven days after instrumentation. Microbiological samples were grown in culture medium and incubated at 37°C for 48 hours. Results: The results were submitted to the Kruskal-Wallis and Dunn (5%) statistical tests. NaOCl and CHX completely eliminated the microorganisms of the root canals. Castor oil and ginger significantly reduced the number of CFU of the tested bacteria. Reduction of CFU/mL at the 1st and 2nd collections for groups G1, G2, G3 and G4 was greater in comparison to groups G5 and G6. Conclusion: It was concluded that 2.5% sodium hypochlorite and 2% chlorhexidine gel were more effective in eliminating C. albicans and E. faecalis, followed by the castor oil and glycolic ginger extract. The Aloe vera extract showed no antimicrobial activity. .

Aderência de Candida albicans em resina acrílica termicamente ativada e liga de cobalto cromo in vitro / Adherence of Candida albicans in thermally activated acrylic resin and cobalt chromium alloy in vitro

O objetivo deste trabalho foi verificar in vitro a adesão de Candida albicans às superfícies de resina acrílica quimicamente ativada e liga de cobalto cromo. Foram confeccionados vinte e sete corpos de prova em forma retangular de três tipos: um grupo de resina acrílica termicamente ativada (n=9), um grupo de cobalto cromo (n=9) e um grupo com metade de sua extensão composta por resina acrílica termicamente ativada e metade por liga de cobalto cromo (n=9). Dois mililitros da cultura de Candida albicans foram adicionados em trinta poços de duas placas para cultura de células. Os corpos de prova foram imersos na posição vertical, individualmente, em cada poço contendo cultura de Candida albicans e mantidos em incubação por 48 horas. Após este período, foram obtidas diluições seriadas da cultura que foram semeadas em placa contendo ágar Sabouraud (em duplicata), incubadas a 37o C por 48h e as unidades formadoras de colônias contadas. Os corpos de prova foram removidos e passados em solução fisiológica (NaCl 0,9%) esterilizada contendo pérolas de vidro que foram agitadas em agitador de tubos Vortex por 1 minuto, após agitação alíquotas de 0,1mL e diluições decimais foram dispensadas em placas de petri contendo ágar Sabouraud. As placas foram incubadas a 370C por 48h. Após o crescimento, as unidades formadoras de colônias por placa foram contadas e as UFC/mL calculadas. Concluíram que leveduras da espécie Candida albicans colonizam resina acrílica e liga metálica em cobalto cromo in vitro e que a liga metálica em cobalto cromo foi mais colonizada por Candida albicans quando comparada à resina acrílica

The objective of this study was to substantiate the adherence of in vitro Candida albicans to the surfaces of a chemically activated acrylic resin and cobalt chromium alloy. There were created twenty-seven test pieces in rectangular shape of three types: a group of heat- -activated acrylic resin (n = 9), a group of cobalt-chromium (n = 9), and a group with half of its length made of acrylic resin thermally activated and half by cobalt-chromium (n = 9). Two milliliters of the culture of Candida albicans were added in thirty wells of two plates for cell culture. The test pieces were immersed vertically in individual well containing culture of Candida albicans and kept in incubation for 48 hours. After this period, serial dilutions of the culture were spread in plate of Sabouraud agar (in duplicate), incubated at 37C for 48h and the colony forming units counted. The test pieces were removed and spread in sterile saline (0.9% NaCl) containing glass beads that were shaken in Vortex tubes for 1 minute, after aliquots shaking of 0.1 mL and decimal dilutions were dispensed in petri plates containing Sabouraud agar. The plates were incubated at 37C for 48 hours. After growing the colony forming units per plate were counted and the CFU/mL calculated. They concluded that Candida albicans colonizes acrylic resin and cobalt-chromium alloy in vitro and that cobalt chromium alloy was more colonized by Candida albicans when compared with acrylic resin

Efeitos do suco de laranja e da molhabilidade da cerâmica sobre a formação do in situ / The effects of orange juice and ceramic wettability on in situ biofi lm formation

Este estudo procurou determinar os parâmetros do biofilme formado in situ, em cerâmica de revestimento, de acordo com a disponibilidade de carboidrato (suco de laranja) e a molhabilidade da superfície cerâmica. Discos de duas cerâmicas foram feitos (Vita VM7 e VM13, Vita Zahnfabrik, Bad Sackingen, Alemanha). Quatro discos foram aderidos a dispositivos palatais usados por oito voluntários. Colocou-se três gotas da água destilada (pH=6) ou suco de laranja (pH=3,5) sobre os discos oito vezes/dia. Após 48 horas in situ, os dois discos mais palatais foram removidos do dispositivo e analisados com microscopia confocal de varredura a laser (MCVL) para caracterização do biofilme (espessura média, μm e biovolume, μm3/μm2). O ângulo de contato (°) foi medido nas superfícies polidas de espécimes controle, e após 15 dias de uso nos espécimes da região frontal do dispositivo palatal. Os dados foram submetidos à estatística descritiva e inferencial (Mann-Whitney e teste t, p < 0,05). A molhabilidade de ambas as cerâmicas não diferiu estatisticamente. A formação do biofilme sobre as cerâmicas de revestimento não foi influenciada pelo suco de laranja ou pela molhabilidade da cerâmica...

This study aimed to determine the in situ biofilm parameters on veneering ceramics, according to the carbohydrate (orange juice) availability and the ceramic surface wettability. Disks were made out of two veneering ceramics (Vita VM7 and VM13, Vita Zahnfabrik, Bad Sackingen, Alemanha). Four disks were bonded to palatal devices worn by eight volunteers. The subjects dripped three drops of distilled water (pH=6) or orange juice (pH=3.5) on the disks 8x/day. After 48 h in situ, the two palatal disks were removed from the device and analyzed by Confocal Laser Scanning Microscopy (CLSM) for biofilm characterization (mean thickness, μm and biovolume, μm3/ μm2). The contact angle (°) was measured on the polished surfaces of as sintered specimens and after 15 day of in situ usage, on the frontal disks of the palatal device. The data were subjected to descriptive and inferential (Mann-Whitney and paired-t test, at p < 0.05) statistics. The biofi lm parameters of VM13 were not measurable, whereas the VM7 biofilm parameters were not signifi cantly affected by the type of liquid. The wettability of both ceramics also did not differ statistically. The biofi lm formation on the veneering ceramics was neither influenced by the orange juice dripping nor the ceramics’ wettability...

Antifungal effect of plant extracts on Candida albicans biofilm on acrylic resin / Efeito antifúngico de extratos vegetais sobre biofilme de Candida albicans em resina acrílica

Objetivo: Avaliar potencial antifúngico dos extratos de Equisetum arvense L. (cavalinha), Glycyrrhiza glabra L. (alcaçuz), Punica granatum L. (romã) e Stryphnodendron barbati-mam Mart. (barbatimão) sobre biofilme de Candida albicans em resina acrílica. Material e métodos: Cepa-padrão de C. albicans foi cultivada em ágar Sabouraud-dextrose por 24 h a 37°C. Após padronização do inóculo (106 células/mL) em espectrofotômetro, foram mantidos em caldo Brain Heart Infusion suplementado comsacarose (5%) um disco de resina acrílica estéril com 100 μL do inóculo padronizado, por 5 dias a 37 °C. As amostras dos grupos tratados (n = 10) foram expostas separadamente à concentração de 50 mg/mL de cada extrato por 5 min e ao antifúngico nistatina (48.83 UI/mL). Para o grupo não tratado (controle, n = 10) foi utilizada solução fisiológica estéril (NaCl 0,9%). Os biofilmes foram desagregados dos discos de resina acrílica por homogeneizador ultrassônico por 30 s. Após diluições decimais, foram feitas semeaduras em placas de Sabouraud-dextrose e incubação por 48 h a 37 ºC. Posteriormente, foram contadas as UFC/mL e os valores foram convertidos em log10 e realizada análise estatística (ANOVA e Tukey Test; p ≤ 0,05). Resultados: Todos os extratos naturais e a nistatina proporcionaram reduções significativas (p < 0,01) do biofilme de C. albicans em comparação ao grupo controle (NaCl 0,9%), no entanto, não houve diferença estatística entre os extratos (p = 0,1567). Conclusões: Houve formação de biofilme de C. albicans em resina acrílica e todos os extratos vegetais foram efetivos para esta levedura, atuando semelhantemente à nistatina.

Objective: Evaluating the antifungal potential of Equisetum arvense L. (horsetail), Glycyrrhiza glabra L. (licorice), Punica granatum L. (pomegranate) and Stryphnodendron barbatimam Mart. (barbatimão) extracts, after Candida albicans biofilm formation on acrylic resin. Material and methods: C. albicans standard strain was cultured on Sabourauddextrose agar for 24 h at 37 °C. After standardized in a spectrophotometer, 100 μL of the inoculums (106 cells/mL) and a sterile acrylic resin disc were maintained in Brain Heart Infusion broth supplemented with sucrose (5%), for 5 days at 37ºC. The samples of the treated groups (n = 10) were separately exposed to a concentration of 50 mg/mL of each extract for 5 minutes or to nystatin (48.83 IU/mL). For the untreated group (control, n = 10), it was used sterile saline (0.9% NaCl). Biofilms were disaggregated from the acrylic resin discs by an ultrasonic homogenizer for 30 s. After decimal dilutions, sowings in Sabouraud-dextrose plates were made with incubation for 48 h at 37°C. Later, CFU/mL was verified and the values were converted to log10 and they had their statistical analysis done (ANOVA and Tukey Test, p ≤ 0.05). Results: It was found that all plant extracts and nystatin resulted in significant reduction of C. albicans biofilm (p < 0.01) compared to the control group (0.9% NaCl). However, all of them showed similar reductions to each other (p = 0.1567). Conclusion: There was biofilm formation of C. albicans on acrylic resin and all plant extracts were effective against this yeast, acting similarly to nystatin.

Effect of Zingiber officinale and propolis on microorganisms and endotoxins in root canals

The purpose of this study was to evaluate the effectiveness of glycolic propolis (PRO) and ginger (GIN) extracts, calcium hydroxide (CH), chlorhexidine (CLX) gel and their combinations as ICMs (ICMs) against Candida albicans,Enterococcus faecalis, Escherichia coli and endotoxins in root canals. Material and Methods: After 28 days of contamination with microorganisms, the canals were instrumented and then divided according to the ICM: CH+saline; CLX, CH+CLX, PRO, PRO+CH; GIN; GIN+CH; saline. The antimicrobial activity and quantification of endotoxins by the chromogenic test of Limulus amebocyte lysate were evaluated after contamination and instrumentation at 14 days of ICM application and 7 days after ICM removal. Results and Conclusion: After analysis of results and application of the Kruskal-Wallis and Dunn statistical tests at 5% significance level, it was concluded that all ICMs were able to eliminate the microorganisms in the root canals and reduce their amount of endotoxins; however, CH was more effective in neutralizing endotoxins and less effective against C. albicans and E. faecalis, requiring the use of medication combinations to obtain higher success.

Effects of probiotic bacteria on Candida presence and IgA anti-Candida in the oral cavity of elderly

Imbalance in the resident microbiota may promote the growth of opportunistic microorganisms, such as yeasts of Candida genus and the development of diseases, especially in aged people. This study evaluated whether the consumption of the probiotic Yakult LB® (Lactobacillus casei and Bifidobacterium breve) was able to influence on the specific immunological response against Candida and on the presence of these yeasts in the oral cavity of 42 healthy aged individuals. Saliva samples were collected before and after the probiotic use for 30 days, 3 times a week. The samples were plated in Dextrose Saboraud Agar with chloramphenicol, the colony-forming units (CFU/mL) were counted and the Candida species were identified. Anti-Candida IgA analysis was conducted using the ELISA technique. ANOVA and Student's t-test were used for normally distributed data and the Wilcoxon test was used for data with non-normal distribution (α=0.05). The results showed a statistically significant reduction (p<0.05) in Candida prevalence (from 92.9% to 85.7%), in CFU/mL counts of Candida and in the number of non-albicans species after consumption of the probiotic. Immunological analysis demonstrated a significant increase (p<0.05) in anti-Candida IgA levels. In conclusion, probiotic bacteria reduced Candida numbers in the oral cavity of the elderly and increased specific secretory immune response against these yeasts, suggesting its possible use in controlling oral candidosis.

Desequilíbrios na microbiota residente podem promover o crescimento de microrganismos oportunistas, como as leveduras do gênero Candida, e o desenvolvimento de doenças, especialmente na população idosa. Este estudo investigou se o consumo do probiótico Yakult LB® (Lactobacillus casei e Bifidobacterium breve) era capaz de influenciar na resposta imune secretória anti-Candida e na presença destes microrganismos na cavidade bucal de 42 idosas saudáveis. Amostras de saliva foram coletadas antes e depois do consumo do probiótico por 30 dias, 3 vezes por semana. As amostras foram semeadas em Agar Saboraud Dextrose com cloranfenicol, as unidades formadoras de colônias (UFC/mL) foram contadas e as espécies de Candida foram identificadas. A análise de IgA anti-Candida foi realizada pela técnica ELISA. Os resultados demonstraram redução na prevalência de Candida (de 92,9% para 85,7%), na contagem de UFC/mL (p≤0,05) e no número de espécies não-albicans, depois do consumo do probiótico. As análises imunológicas mostraram um aumento significativo dos níveis de IgA anti-Candida (p≤0,05). Concluindo, as bactérias probióticas reduziram significantemente a quantidade de Candida na cavidade bucal dos idosos e aumentaram a resposta imune secretória específica para esta levedura, sugerindo a possibilidade de sua utilização no controle da candidose bucal.

Efeitos clínicos e radiográficos do laser em baixa intensidade após a extração de terceiros molares inclusos / Clinical and radiographic effects of low intensity laser after extraction of unerupted third molars

Introdução: O laser em baixa intensidade tem sido indicado como tratamento coadjuvante no pós-operatório da cirurgia de extração dentária. Objetivo: O objetivo deste trabalho foi avaliar os efeitos clínicos e radiográficos do laser em baixa intensidade na cirurgia de exodontia de terceiros molares inclusos. Material e método: Oito pacientes foram submetidos à extração dos terceiros molares inferiores inclusos. O dente esquerdo foi tratado com laser durante a cirurgia e por mais dois dias do pós-operatório (Grupo Laser). A cirurgia do dente direito foi realizada após 15 dias da cirurgia do dente esquerdo e não recebeu laserterapia (Grupo Controle). A avaliação clínica do pós-operatório foi baseada na medida do edema e na análise de questionário para avaliação da dor. Após 40 dias de cada cirurgia, foram feitas radiografias periapicais digitais para medida das densidades ópticas da reparação óssea, por meio do programa Image J. Os dados obtidos na medida do edema e na análise de densidade óptica foram submetidos ao teste estatístico t de Student. Resultado: O nível de dor dos pacientes no pós-operatório foi menor no Grupo Laser em relação ao Grupo Controle. Entretanto, na medida do edema e na análise de densidade óptica das radiografias, não houve diferença estaticamente significante do Grupo Laser em relação ao Grupo Controle. Conclusão: De acordo com os parâmetros utilizados neste estudo, concluiu-se que a aplicação do laser em baixa intensidade promoveu analgesia no pós-operatório, porém não teve efeito sobre o edema e a reparação óssea.

Introduction: The low intensity laser therapy (LLLT) has been indicated as coadjuvant treatment of postoperative dental extraction surgery. Objective: The aim of this study was to evaluate the clinical and radiological findings of LLLT in the surgery for extraction of unerupted third molars. Material and method: Eight patients were submitted to extraction of mandibular third molar. The left tooth was treated with laser during surgery and for another 2 days after surgery (Laser Group). The right tooth surgery was performed after 15 days and did not receive laser therapy (control group). Clinical evaluation of the postoperative period was based on the measuring of edema and analysis of a questionnaire to assess pain. After 40 days of each surgery, digital periapical radiographs were made and measured the optical density of bone repair were analyzed using the Image J. The data obtained in the measurement of edema and analysis of optical density were tested using Student t test. Result: The level of pain in postoperative patients was lower in the laser group compared to the control group. However, in the measurement of edema and analysis of optical density of radiographs there was no statistically significant difference in the laser group compared to control. Conclusion: According to the parameters used in this study, we can concluded that the application of LLLT promoted analgesia postoperatively, but did not show effects on edema and bone repair.