ABSTRACT Objective: To evaluate an imaging protocol for use as a diagnostic and calibration tool for dentists before and after practical activity. Material and Methods: Thirty photos of children's teeth with or without changes in dental enamel were selected and evaluated by a group of experienced dentists previously calibrated to establish the diagnosis defined as the gold standard. After instructions, the images were shown to a group of postgraduate dentists for free identification of dental changes. Subsequently, a lecture on molar incisor hypomineralization (MIH) was carried out, and, at 14 days and all calibration was performed using the criteria previously. The retest was performed at 28 days. After experience in clinical activity in the following two weeks, the post-test was performed at 49 days. Data were analyzed using Cohen's kappa coefficient. Results: Theoretical learning on the subject showed low inter-examiner agreement when the diagnosis of defects was made from images obtained from intraoral photographs. After clinical practice, there was greater intra-examiner agreement. After theoretical training, dentists started to identify different types of enamel alteration, although with low agreement between them. Conclusion: Clinical experience in theoretical and imaging training favored the identification of defects. However, it is necessary to improve the protocol to establish a reliable and viable diagnostic method for calibration in MIH.

ABSTRACT Objective: To investigate the types of restorative materials used for restorative treatment in primary teeth through a retrospective university-based study. Material and Methods: The sample consisted of all clinical records of children attended at the Pediatric Dentistry Clinic at the School of Dentistry of Ribeirão Preto at the University of São Paulo in Brazil. Inclusion criteria were primary anterior and posterior teeth that received dental restorations for treatment of dental caries lesions, dental trauma or dental development defects from 2013 to 2018. Restoration repairs and interim restorations during this period were also recorded. Descriptive analyzes were performed to assess the distribution according to the type of restorative material used over the years. Results: A total of 5,236 restorative procedures were performed in primary teeth, including restoration repair and interim restorations. Of those, 69% were done in posterior teeth and 31% in anterior teeth. Sixty percent of the procedures performed during this period were made of composite resin and a lower percentage of glass ionomer cement (18%) followed by silver amalgam (1%). The number of interim restorations was smaller but proportional to those of composite resin over the years. Conclusion: A tendency to carry out restorative treatment of primary teeth with composite resin during the 6 years of follow-up was observed.

ABSTRACT Objective: To verify, through clinical and radiographic evaluations, the in vivo response of the dentin-pulpal complex of human primary teeth after pulpotomy with MTA and Biodentine™ in a follow-up period of 3, 6, and 12 months. Material and Methods: Thirty teeth were divided into MTA pulpotomy (n = 15) and Biodentine™ pulpotomy (n = 15) from children between 5 and 9 years of age, a randomized clinical trial with simple random sampling. The materials were inserted into the cavity after opening and removing the coronary pulp tissue. The cavity base consisted of glass ionomer cement and light-cured composite resin restoration. Clinical and radiographic analyses were performed after 3, 6, and 12 months. Statistical analysis by Fisher's exact test for dichotomous data at a 5% significance level was utilized. Results: Both materials caused color change after 12 months. However, MTA showed a higher percentage than Biodentine™ (p<0.0001). Pain was detected only with Biodentine™ at six months and mobility at 12 months (p=0.0013). Radiographically, after 12 months, periapical lesions, interradicular lesions, and internal resorption were evidenced in 13% of the cases for Biodentine™-treated teeth (p<0.0013). MTA induced pulp calcification in 13% of cases, unlike Biodentine™ (p<0.0013). Conclusion: BiodentineTM and MTA are suitable for clinical use in pulpotomy treatment, yet both materials lead to tooth discoloration.

Abstract The objective of this study was to compare the activation of gelatinases in dentin-enamel junction (DEJ) and underlying dentin of permanent teeth after experimental radiotherapy in conventional and hypofractionated modalities. Newly extracted third molars (n = 15) were divided into three experimental radiotherapy groups: control, conventional (CR), and hypofractionated (HR) (n = 5 per group). After in vitro exposure to ionizing radiation, following standardized protocols for each modality, a gelatinous substrate was incubated on the tooth slices (n = 10 per group). Activation of gelatinases was measured by in situ zymography, expressed in arbitrary fluorescence units (mm2) from three tooth regions: cervical, cuspal, and pit. Fluorescence intensity was compared among radiotherapy protocols and tooth regions in each protocol, considering a significance level of 5%. Considering all tooth regions, the fluorescence intensity of the CR group was higher than the HR and control groups, both in DEJ and underlying dentin (p <0.001). In addition, the fluorescence intensity was higher in underlying dentin when compared to DEJ in all groups (p <0.001). Considering each tooth region, a statistically significant difference between CR and HR was only observed in the pit region of underlying dentin (p <0.001). Significant and positive correlations between fluorescence intensities in DEJ and underlying dentin were also observed (p <0.001). Experimental radiotherapy influenced the activation of gelatinases, as well as exposure to the conventional protocol can trigger a higher activation of gelatinases when compared to hypofractionated, both in DEJ and underlying dentin.

Resumo O objetivo deste estudo foi comparar a ativação de gelatinases na junção dentina-esmalte (DEJ) e na dentina subjacente de dentes permanentes após a radioterapia experimental nas modalidades convencional e hipofracionada. Os terceiros molares recém-extraídos (n = 15) foram divididos em três grupos de radioterapia experimental: controle, convencional (CR) e hipofracionada (HR) (n = 5 por grupo). Após a exposição in vitro à radiação ionizante, seguindo protocolos padronizados para cada modalidade, um substrato gelatinoso foi incubado nas fatias de dente (n = 10 por grupo). A ativação das gelatinases foi medida por zimografia in situ, expressa em unidades arbitrárias de fluorescência (mm2) de três regiões do dente: cervical, cúspide e fossa. A intensidade da fluorescência foi comparada entre os protocolos de radioterapia e as regiões do dente em cada protocolo, considerando um nível de significância de 5%. Considerando todas as regiões do dente, a intensidade de fluorescência do grupo CR foi maior do que a dos grupos HR e controle, tanto no DEJ quanto na dentina subjacente (p <0,001). Além disso, a intensidade da fluorescência foi maior na dentina subjacente quando comparada à DEJ em todos os grupos (p <0,001). Considerando cada região do dente, uma diferença estatisticamente significativa entre CR e HR foi observada apenas na região da fossa da dentina subjacente (p <0,001). Também foram observadas correlações significativas e positivas entre as intensidades de fluorescência no DEJ e na dentina subjacente (p <0,001). A radioterapia experimental influenciou a ativação das gelatinases, assim como a exposição ao protocolo convencional pode desencadear uma maior ativação das gelatinases quando comparada ao hipofracionamento, tanto no DEJ quanto na dentina subjacente.

Resumo O objetivo deste trabalho foi Investigar a formação de osteoclastos in vivo e se o leucotrieno B4 (LTB4) incorporado em microesferas (MS) poderia ser usado como estratégia terapêutica para promover uma entrega sustentada do mediador e prevenir a diferenciação dos osteoclastos. Métodos: In vivo, a periodontite apical foi induzida em camundongos para investigar a diferenciação e sinalização de osteoclastos na ausência de 5-lipoxigenase (5-LO). In vitro, LTB4-MS foi preparado usando um processo de evaporação e extração de solvente de emulsão de óleo em água. A caracterização e a eficiência do encapsulamento do LTB4 foram investigadas. Macrófagos J774A.1 foram cultivados na presença de fator estimulador de colônia de monócitos (M-CSF) e ligante para o receptor ativador do fator nuclear kappa B (RANKL) e, então, estimulados com LTB4-MS. Citotoxicidade, captação in vitro de MS-LTB4, formação de osteoclastos e expressão gênica foram avaliadas. Resultados: A via 5-LO regula negativamente a formação de osteoclastos in vivo durante o desenvolvimento da periodontite apical. In vitro, LTB4-MS foram fagocitadas pelos macrófagos e não foram citotóxicos para as células. LTB4-MS inibiu a formação de osteoclastos e a síntese dos genes pró-osteoclastogênicos Acp5, Mmp9, Calcr e Ctsk. Conclusões: LTB4-MS inibiu a diferenciação de macrófagos em um fenótipo osteoclástico e a ativação celular sob estímulo de M-CSF e RANKL.

Abstract To investigate osteoclast formation in vivo and if leukotriene B4 (LTB4) loaded in microspheres (MS) could be used as a therapeutical strategy to promote a sustained delivery of the mediator and prevent osteoclast differentiation. Methods: In vivo, apical periodontitis was induced in mice to investigate osteoclast differentiation and signaling in absence of 5-lipoxygenase (5-LO). In vitro, LTB4-MS were prepared using an oil-in-water emulsion solvent extraction-evaporation process. Characterization and efficiency of LTB4 encapsulation were investigated. J774A.1 macrophages were cultured in the presence of monocyte colony-stimulating factor (M-CSF) and ligand for receptor activator of nuclear factor kappa B (RANKL) and then stimulated with LTB4-MS. Cytotoxicity, in vitro MS-LTB4 uptake, osteoclast formation and gene expression were measured. Results: We found that 5-LO negatively regulates osteoclastic formation in vivo during apical periodontitis development. In vitro, LTB4-MS were up-taken by macrophages and were not cytotoxic to the cells. LTB4-MS inhibited osteoclast formation and the synthesis of osteoclastogenic genes Acp5, Mmp9, Calcr and Ctsk. LTB4-MS inhibited differentiation of macrophages into an osteoclastic phenotype and cell activation under M-CSF and RANKL stimulus.

Resumo O desenvolvimento, estabelecimento e reparo da periodontite apical (PA) depende de vários fatores, que incluem a susceptibilidade do hospedeiro, infecção microbiana, resposta imune, bem como a qualidade do tratamento do canal radicular e a capacidade de reparo do organismo. A compreensão das contribuições genéticas para o risco de desenvolver a PA e apresentar PA persistente tem sido extensivamente explorada na Endodontia moderna. Assim, este manuscrito pretende fornecer uma revisão da literatura em relação aos mediadores bioquímicos envolvidos na sinalização da resposta imune, osteoclastogênese e neoformação óssea, bem como os componentes genéticos envolvidos no desenvolvimento e reparo da PA. Uma revisão narrativa da literatura foi realizada através de uma pesquisa nas bases PUBMED/MEDLINE e uma pesquisa manual nos principais livros sobre a PA. O conhecimento sobre as células, receptores e moléculas envolvidas na resposta imuno-inflamatória do hospedeiro durante a progressão da PA somado ao conhecimento da biologia óssea, especialmente o papel dos osteoblastos, osteócitos e osteoclastos no turnover ósseo, permite a identificação de fatores inerentes ao hospedeiro que podem interferir tanto na progressão como no reparo destas lesões. Os principais resultados dos estudos avaliados na revisão que investigaram a correlação entre polimorfismos genéticos e PA, nos últimos cinco anos, demonstram que os fatores genéticos do indivíduo estão envolvidos no sucesso do tratamento do canal radicular. A discussão desta revisão fornece subsídios que podem ajudar a vislumbrar o desenvolvimento de novas terapias baseadas na identificação de alvos terapêuticos e no desenvolvimento de materiais e técnicas destinadas a atuar a nível molecular para o sucesso clínico, radiográfico e histológico do tratamento endodôntico.

Abstract The development, establishment and repair of apical periodontitis (AP) is dependent of several factors, which include host susceptibility, microbial infection, immune response, quality of root canal treatment and organism's ability to repair. The understanding of genetic contributions to the risk of developing AP and presenting persistent AP has been extensively explored in modern Endodontics. Thus, this article aims to provide a review of the literature regarding the biochemical mediators involved in immune response signaling, osteoclastogenesis and bone neoformation, as the genetic components involved in the development and repair of AP. A narrative review of the literature was performed through a PUBMED/MEDLINE search and a hand search of the major AP textbooks. The knowledge regarding the cells, receptors and molecules involved in the host's immune-inflammatory response during the progression of AP added to the knowledge of bone biology allows the identification of factors inherent to the host that can interfere both in the progression and in the repair of these lesions. The main outcomes of studies evaluated in the review that investigated the correlation between genetic polymorphisms and AP in the last five years, demonstrate that genetic factors of the individual are involved in the success of root canal treatment. The discussion of this review gives subsides that may help to glimpse the development of new therapies based on the identification of therapeutic targets and the development of materials and techniques aimed at acting at the molecular level for clinical, radiographic and histological success of root canal treatment.

Resumo Considerando que a fumaça de cigarro é um problema de saúde pública que está crescendo entre os adolescentes, o objetivo deste estudo foi investigar o impacto da fumaça de cigarro na sinalização osteogênica e osteoclastogênica da sutura palatina mediana de ratos. Foram utilizados ratos Wistar machos expostos (n=30) e não expostos à fumaça de cigarro (n=30). A exposição à fumaça de cigarro foi realizada duas vezes ao dia por 3 minutos, com um intervalo de 12 horas entre as exposições. Os animais foram mortos após o período experimental de 3, 7, 14 e 21 dias. Os tecidos coletados foram analisados em microscópico de luz e pelo RT-PCR em tempo real foi realizado para investigar a expressão gênica. s dados obtidos foram comparados usando o testes de Kruskal Wallis e Dunn (⍺ = 5%). Morfoligicamente, não houve mudança significativa na sutura palatina mediana nos ratos expostos ou não à fumaça de cigarro durante os tempo de 3, 7, 14 e 21 dias (p> 0.05). Por outro lado, a sinalização osteogênica esta aumentada nos animais expostos à fumaça e foi caracterizado por um aumento da produção de RANKL aos 3 e 14 dias (p <0.05), sem mudança na síntese da produção de RANK e osteoprotegerina (p> 0.05). Curiosamente, nos animais expostos, também foi observado um aumento precoce da síntese de osteocalcina, sialoproteína óssea e de osteopontina aos 3 dias de exposição, o que não foi mantido ao longo do tempo. A fumaça de cigarro modula a sinalização osteogênica e osteoclastogênica na sutura palatina mediana de ratos jovens, apesar de não tenha sido evidenciado alterações morfológicas.

Abstract Considering that smoking is a public health problem that has been growing among adolescents, the aim of this study was to investigate the impact of cigarette smoke on osteogenic and osteoclastogenic signaling in middle palatal suture of rats. Male Wistar rats exposed (n = 30) or not to cigarette smoke (n = 30) were used. Exposure to smoke was carried out for two daily periods of 3 minutes each, with an interval of 12 hours between exposures. After the experimental periods of 3, 7, 14 and 21 days, the animals were euthanized. The collected tissues were analyzed using light microscopy and real-time RT-PCR was performed to investigate gene expression. The data obtained were compared using the Kruskal Wallis and Dunn tests (⍺ = 5%). Morphologically, there were no significant changes in the middle palatal suture of rats exposed or not to cigarette smoke during 3, 7, 14 and 21 days (p> 0.05). On the other hand, osteoclastogenic signaling was increased in animals exposed to smoke and was characterized by a higher production of RANKL at 3 and 14 days (p <0.05), with no change in the synthesis of RANK and osteoprotegerin (p> 0.05). Interestingly, in the exposed animals, an early increase in the synthesis of osteocalcin, bone sialoprotein and osteopontin was also identified at 3 days of exposure (p <0.05), not sustained over time (p> 0.05). Cigarette smoke modulates osteogenic and osteoclastogenic signaling in the middle palatal suture of young rats, although morphological changes have not been evidenced.

Resumo Objetivo: Avaliar a resposta do tecido conjuntivo subcutâneo de camundongos isogênicos expostos à Biodentine™ e ao Trióxido Mineral Agregado (MTA). Métodos: Um total de 120 camundongos foram divididos em 4 grupos e 3 períodos experimentais diferentes (7, 21 e 63 dias): Biodentine™ (Septodont, Saint Maur des Fosses, França); Pro-Root MTA (Dentsplay, Tulsa, EUA); óxido de zinco eugenol (Biodinâmica Química e Farmacêutica LTDA., Ibiporã, PR - Brasil); e controle negativo - Sham. Após os períodos experimentais, análises microscópicas descritivas, semiquantitativas e quantitativas do processo inflamatório foram analisadas nos cortes de H&E e ainda, foi realizada a avaliação da expressão gênica de Il10, Infg, Il6, Il1r1 e Tnf (qRT-PCR). Os dados obtidos foram analisados por meio do teste do qui-quadrado e da análise de variância (ANOVA) two-way, seguido do pós-teste de Bonferroni (nível de significância de 5%). Resultados: Na análise microscópica observou-se discreto infiltrado inflamatório, com predomínio de macrófagos esparsos e polimorfonucleares, leve fibrose tecidual, cápsula fibrosa regular e com calcificações distróficas, em todos os grupos que receberam os materiais (Biodentine ™ e Pro-Root MTA). Paralelamente, todos os materiais modulam a expressão gênica das diferentes citocinas e receptores avaliados. Conclusão: Pro-Root MTA e Biodentine™ mostraram compatibilidade tecidual, inflamação mediada, com aumento do tecido fibroso e produção de citocinas pró- e antiinflamatórias.

Abstract The aim of this study was to evaluate the subcutaneous connective tissue response of isogenic mice exposed to tricalcium silicate (Biodentine) and aggregated mineral trioxide (ProRoot MTA). A total of 120 mice were divided into 4 groups in 3 different experimental periods (7, 21 and 63 days): Biodentine; Pro-Root MTA; zinc oxide-eugenol and; Negative control - Sham. After the experimental periods microscopic descriptive, semi-quantitative and quantitative analysis of the inflammatory process were analyzed on H&E sections and evaluation of the gene expression of Il10, Infg, Il6, Il1r1 and Tnf (qRT-PCR) were performed. The data obtained were analyzed using the chi-square test and two-way analysis of variance (ANOVA) followed by the Bonferroni post-test (5% significance level). Results: In the microscopic analysis, a slight inflammatory infiltrate was observed, with a predominance of sparse macrophages and polymorphonuclear cells, slight tissue fibrosis, regular fibrous capsule and with dystrophic calcifications, in all groups that received the materials (Biodentine and Pro-Root MTA). In parallel, all materials modulated the gene expression of the different cytokines and receptors evaluated. Conclusion: Pro-Root MTA and Biodentine showed a tissue compatibility, mediated inflammation, with increased fibrous tissue and production of pro- and anti-inflammatory cytokines.

Abstract: In this study, we evaluated the knowledge, attitudes, and psychosocial impacts among Brazilian pediatric dentists during the COVID-19 pandemic. A cross-sectional study with primary data collection was carried out using an online structured questionnaire. Data were submitted to descriptive analysis by using absolute and relative frequencies. A chi-square test was used for association analysis and log-linear regression models to estimate the prevalence ratio (5%). The population comprised mostly women and knowledge regarding COVID-19 was satisfactory (above 80% in most items). N95 masks and faceshields were used, albeit 64.22% reported difficulty in providing dental care to children due to the need for extra Personal Protective Equipment. Thirty eight percent provided urgency/emergency dental care, 59.78% performed invasive procedures, 59.56% used high speed handpieces, 8.44% started using cariostatics, and 6.22% introduced the use of chemomechanical caries removal agents. Fear for the future (PR = 1.21) and use of medications (for anxiety, depression, or insomnia) increased (PR = 1.16) among the ones who had wage losses. Brazilian pediatric dentists have knowledge about COVID-19, and attitudes in their clinical routines changed due the pandemic. Financial life was harmed and a negative impact of the pandemic in psychosocial aspects of workers was found.

ABSTRACT Objective To evaluate the physical conditions and presence of residues of toothbrushes used by mothers and their babies and mothers' knowledge about toothbrush care. Material and Methods This was a cross-sectional study comprising a convenience sample represented by 60 mother-baby pairs. The mothers answered a questionnaire to evaluate their knowledge of toothbrush care. A calibrated dentist performed a visual inspection of the toothbrushes. Statistical analysis was performed using the chi-square and Fisher’s exact tests, with a significance level of 5%. Results It was found that 82% of the mothers had never received instructions regarding the care of toothbrushes after use (p=0.024). Most of them believed that their toothbrushes (70%) and their children’s toothbrushes (88%) were in good condition to use (p=0.043). However, most mother’s toothbrushes presented an unacceptable deformity of the bristles (65%) and the presence of residues (60%). In addition, babies’ toothbrushes also presented unacceptable deformities of the bristles (52%) and residues (55%). There was an association between the lack of instructions received by the mother and the presence of deformity and residues on the mother’s toothbrush bristles (p=0.037 and p=0.003, respectively). Conclusion Most mothers had never received instructions regarding toothbrush care, which is reflected in the condition of their and their baby’s toothbrushes, which presented unacceptable physical conditions concerning deformation and presence of residues.

Abstract Molar incisor hypomineralization (MIH) is often accompanied by dental hypersensitivity and difficulty in achieving effective analgesia. Objective: This study evaluated the effectiveness of preemptive analgesia in children with severe MIH, post-eruptive enamel breakdown, and hypersensitivity. Methodology: Ibuprofen (10 mg/kg child weight) or placebo was administered, followed by infiltrative anesthesia and restoration with resin composite. Hypersensitivity was evaluated in five moments. The data were analyzed using the chi-square test, Fisher’s exact test, and t-test. Results: Preemptive analgesia provided benefits for the treatment of severe cases of MIH, with an increase in the effectiveness of infiltrative anesthesia and improved patient comfort during the restorative procedure. Conclusion: Preemptive analgesia has shown efficacy in reducing hypersensitivity during restorative dental procedures, evidencing the significance of this study for patients with MIH and hypersensitivity.

Abstract Objective: To evaluate the periapical healing following root canal treatment in teeth with apical periodontitis (in vivo) and the cytotoxic potential of root canal sealers in vitro. Material and Methods: Apical periodontitis was induced in 60 dogs' teeth and root canals were filled with Sealapex (40 roots), EndoREZ (40 roots), intracanal dressing (20 roots), or left untreated (20 roots). After 30 and 90 days, histopathological analyses were made. In vitro, J774.1 macrophages were stimulated with root canal sealers extracts, cytotoxicity was assessed using lactate dehydrogenase assay, and qRT-PCR was used to analyze TNF-α gene expression. Results: In vivo, smaller apical periodontitis and lower inflammatory cell infiltrate were found in teeth treated with Sealapex compared to EndoREZ. In vitro, EndoREZ was cytotoxic and induced TNF-α gene expression by macrophages differently from Sealapex. Conclusion: Sealapex allowed improved tissue repair following root canal treatment in teeth with apical periodontitis compared to EndoREZ. Synthesis of TNF-α induced by LPS was enhanced by EndoREZ, whereas Sealapex prevented pro-inflammatory gene expression.

Resumo O objetivo do presente estudo in vivo foi avaliar a contaminação bacteriana de protetores bucais esportivos, a rugosidade da superfície e a eficácia do spray de gluconato de clorexidina na desinfecção desses dispositivos. Um ensaio clínico randomizado, cego, cruzado foi realizado com vinte crianças de 9 a 13 anos, que praticavam artes marciais, participaram de todas as fases do estudo. As crianças foram orientadas a usar o protetor bucal por 3 dias alternados durante 1 hora e, após o uso, borrifar água de torneira estéril ou clorexidina 0,12%. Os protetores foram analisados por ensaio MTT, Hibridização DNA-DNA e microscopia confocal a laser antes e após o uso por 2 semanas. Os dados foram analisados pelos teste de Wilcoxon, teste t de Student, e correlação de Pearson, com nível de significância de 5%. Observou-se que os protetores bucais do grupo controle estavam mais contaminados com microrganismos cariogênicos do que os do grupo experimental (clorexidina) (p <0,05). O uso de protetores bucais com spray de clorexidina reduziu significativamente a contaminação bacteriana em relação ao grupo controle (p = 0,007). A rugosidade da superfície dos protetores bucais aumentou significativamente após o uso, independentemente da aplicação de spray de clorexidina. Uma correlação moderada (r = 0,59) foi observada entre a rugosidade da superfície e a contaminação do micro-organismo apenas para o grupo controle. Os protetores bucais esportivos apresentam intensa contaminação microbiana e aumento da rugosidade superficial após o uso. O uso de spray de clorexidina foi eficaz para reduzir a contaminação dos protetores bucais usados por crianças.

Abstract The aim of the present in vivo study was to evaluate the bacterial contamination of sports mouthguards, surface roughness, and the efficacy of chlorhexidine gluconate spray in the disinfection of these devices. A randomized, blinded cross-over clinical trial was performed with twenty 9 to 13 years old children who practiced martial arts and participated in all phases of the study. They were instructed to wear mouthguards 3 alternated days a week for 1 hour and, after use, to spray sterile tap water or chlorhexidine 0.12%. The mouthguards were analyzed by MTT assay, Checkerboard DNA-DNA hybridization, and confocal laser microscopy prior and after use for 2 weeks. Data were analyzed by Wilcoxon and t-Student, and Pearson correlation tests, with 5% significance level. Were observed that mouthguards of the control group were more contaminated with cariogenic microorganisms than those of the chlorhexidine group (p<0.05). The mouthguards use of spray of chlorhexidine reduced significantly the bacteria contamination compared with control group (p = 0.007). The surface roughness of the mouthguards increased significantly after use, irrespective of application of chlorhexidine spray. A moderate correlation (r=0.59) was observed between surface roughness and the cariogenic microorganism’s contamination only for control group. Sports mouthguards had intense microbial contamination and increased surface roughness after its use. The use of chlorhexidine spray was effective for reducing the mouthguards contamination used by children.

Resumo O objetivo deste estudo foi avaliar in vitro a citotoxicidade dos cimentos endodônticos Sealapex Xpress e Real Seal XT pelo ensaio de MTT e a ativação de macrófagos J774.1. Os cimentos endodônticos Sealapex Xpress e Real Seal XT foram pesados e os extratos foram obtidos a partir da diluição em meio de cultura DMEM por 48 horas (10mg/mL, 1mg/m, e 0,1 mg/mL). A viabilidade celular foi avaliada pelo ensaio MTT e a produção de citocinas (TNF-a, IL-6 e IL-10) foi investigada pelo ensaio imunoenzimático (ELISA) em células de linhagem (macrofagos J774.1). Os dados obtidos foram analisados utilizando-se análise de variância de uma via e pós-teste de Tukey (a=0,05). A viabilidade celular após 24 ou 48 horas não foi afetada nas concentrações de 0,1 ou 1 mg/mL dos dois cimentos estudados (p>0,05). Por outro lado, na concentração 10 mg/mL, a viabilidade celular foi significativamente mais baixa (p <0,05). Observou-se que o Sealapex Xpress inibiu a produção de TNF-a, enquanto o Real Seal XT induziu a secreção de TNF-a às 24 h (p<0,05). A produção de IL-6 foi induzida pelo Real Seal XT, mas não pelo Sealapex Xpress (p<0,05). A secreção da citocina anti-inflamatória IL-10 foi induzida tanto pelo Real Seal XT quanto pelo Sealapex Xpress. IL-4 não foi detectada em nenhum grupo. Em conclusão, os dois cimentos obturadores apresentaram baixa toxicidade, mas ativaram os macrófagos de modo distinto. A ativação pelo Sealapex Xpress foi caracterizada pela inibição do TNF-a e indução da IL-10, enquanto o Real Seal XT induziu somente IL-6.

Abstract This study evaluated the cytotoxicity of Sealapex Xpress and Real Seal XT and their effect on macrophage activation. J774.1 macrophages were incubated with Sealapex Xpress and Seal Real XT (0.1, 1.0, and 10 mg/mL) for 24 and 48 h. Cell viability was assessed by the MTT assay and macrophage activation was measured by pro- and anti-inflammatory cytokine production using ELISA. Data were analyzed using one-way ANOVA and Tukey’s post-test (a=0.05). Cell viability was not affected with 0.1 or 1.0 mg/mL of extracts of Sealapex Xpress and Real Seal XT at 24 and 48 h (p>0.05), but was significantly lower when cells were exposed to 10 mg/mL of both sealers (p<0.05). Sealapex Xpress inhibited the production of TNF-a, whereas Real Seal XT induced TNF-a secretion at 24 h (p<0.05). IL-6 production was induced by Real Seal XT, but not by Sealapex Xpress (p<0.05). Real Seal XT and Sealapex Xpress induced the secretion of anti-inflammatory IL-10. IL-4 was not detected in any group. In conclusion, both sealers had low toxicity but differentially activated macrophages. Macrophage activation by Sealapex Xpress was characterized by inhibition of TNF-a and induction of IL-10, whereas Real Seal XT induced IL-6 solely.

Abstract Purpose To evaluate the kinetics of apical periodontitis development in vivo , induced either by contamination of the root canals by microorganisms from the oral cavity or by inoculation of bacterial lipopolysaccharide (LPS) and the regulation of major enzymes and receptors involved in the arachidonic acid metabolism. Methodology Apical periodontitis was induced in C57BL6 mice (n=96), by root canal exposure to oral cavity (n=48 teeth) or inoculation of LPS (10 µL of a suspension of 0.1 µg/µL) from E. coli into the root canals (n= 48 teeth). Healthy teeth were used as control (n=48 teeth). After 7, 14, 21 and 28 days the animals were euthanized and tissues removed for histopathological and qRT-PCR analyses. Histological analysis data were analyzed using two-way ANOVA followed by Sidak’s test, and qRT-PCR data using two-way ANOVA followed by Tukey’s test (α=0.05). Results Contamination by microorganisms led to the development of apical periodontitis, characterized by the recruitment of inflammatory cells and bone tissue resorption, whereas inoculation of LPS induced inflammatory cells recruitment without bone resorption. Both stimuli induced mRNA expression for cyclooxygenase-2 and 5-lipoxygenase enzymes. Expression of prostaglandin E 2 and leukotriene B 4 cell surface receptors were more stimulated by LPS. Regarding nuclear peroxisome proliferator-activated receptors (PPAR), oral contamination induced the synthesis of mRNA for PPARδ, differently from inoculation of LPS, that induced PPARα and PPARγ expression. Conclusions Contamination of the root canals by microorganisms from oral cavity induced the development of apical periodontitis differently than by inoculation with LPS, characterized by less bone loss than the first model. Regardless of the model used, it was found a local increase in the synthesis of mRNA for the enzymes 5-lipoxygenase and cyclooxygenase-2 of the arachidonic acid metabolism, as well as in the surface and nuclear receptors for the lipid mediators prostaglandin E2 and leukotriene B4.