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: The COVID-19 pandemic led to changes in academic teaching and dental education, but the impact on dental professors’ mental health and quality of life remains poorly understood. Thus, the objective of this study was to evaluate the impacts of social distancing and online teaching related to COVID-19 on the quality of life and anxiety of Brazilian dental professors. This was a cross-sectional study conducted from August 2020 to October 2020. Three instruments were used in an online version: a questionnaire about personal data, academic information and online teaching activities, the Generalized Anxiety Disorders 7 (GAD7) scale, and the Abbreviated World Health Organization Quality of Life (WHOQOL-bref) scale. All instruments were sent by e-mail, social media, and messaging apps to private and public universities and professors. Of the 318 responses, 187 (58.8%) were from female professors. Moreover, lack of good internet access and adequate place for online teaching, difficulties in producing teaching materials, and housework roles had a significant impact on the quality of life and anxiety scores (all p-values < 0.05). Also, Brazilian dental professors who declared that they would make greater efforts if the activities were face-to-face had significantly worse quality of life and anxiety scores, and female professors had significantly higher anxiety scores (all p-values < 0.05). These results provide evidence of a negative effect of social distancing and online teaching activities related to COVID-19 outbreak on the health-related quality of life and mental health of Brazilian dental professors.

Resumo O objetivo deste estudo foi avaliar a modulação dos macrófagos M1 e M2 após estímulos com diferentes materiais utilizados durante o tratamento endodôntico. Em cultura de células de macrófagos derivados da medula óssea de camundongos machos C57BL/6 wild-type (WT), após a exposição à cinco cimentos endodônticos: AH Plus, Sealapex Xpress, Endosequence BC Sealer, BioRoot RCS e pasta à base de hidróxido de cálcio foi realizada a análise da expressão gênica dos marcadores para macrófagos M1 e M2 por qRT-PCR (Cxcl10, CxCL9, iNOS, Arg1, Chil3, Retnla e MRC1) e quantificação de citocinas por Luminex® (GM -CSF, IL-10, IL-6, IL-1β e TNF-α). Para valores normais, foi utilizado o teste ANOVA, seguido do pós-teste de Tukey. Para valores não normais, foi utilizado o teste de Kruskall-Wallis. BioRootTM RCS e EndoSequence BC SealerTM estimularam maior expressão de marcadores para macrófagos M1, enquanto a pasta à base de hidróxido de cálcio estimulou expressão mais baixa desses marcadores gênicos. Para o marcador de proteínas para M2, BioRootTM RCS apresentou a maior estimulação, enquanto a pasta à base de hidróxido de cálcio também apresentou menor estimulação. Concluiu-se que os materiais obturadores avaliados aumentaram a expressão genética de marcadores pró- e anti-inflamatórios: TNF-α e IL-10 respectivamente. Os demais marcadores pró inflamatórios mostraram diferenças em relação aos materiais obturadores. No entanto, esse processo não induziu a polarização da resposta inflamatória, resultando em um macrófago híbrido.

Abstract The aim of this study was to evaluate the M1 and M2 macrophage modulation after stimuli with different materials used during endodontic treatment. In bone marrow-derived macrophage cell culture, from males C57BL/6 wild-type (WT) mice, gene expression analysis of markers to M1 and M2 macrophages was performed by qRT-PCR (Cxcl10, CxCL9, iNOS, Arg1, Chil3, Retnla and MRC1) and cytokine quantification by Luminex® (GM-CSF, IL-10, IL-6, IL-1β and TNF-α) after exposure to the five endodontic sealers: AH Plus, Sealapex Xpress, Endosequence BC Sealer, BioRoot RCS and a calcium hydroxide-based paste. For normal values, ANOVA test was used, followed by Tukey post-test. For non-normal values, the Kruskall-Wallis test was used. BioRootTM RCS and EndoSequence BC SealerTM stimulated the highest expression of markers for M1 macrophages, while calcium hydroxide-based paste stimulated the lowest expression of these gene markers. For M2 protein markers, BioRootTM RCS presented the highest stimulation while calcium hydroxide-based paste also presented the lowest stimulation. It was concluded that all the evaluated filling materials increased the genetic expression of pro- and anti-inflammatory markers: TNF-α and IL-10 respectively. The others proinflammatory mediators showed differences against the filling materials. However, this process did not induce the inflammatory response polarization, resulting in a hybrid macrophage.

Resumo O objetivo do presente estudo in vivo foi avaliar o efeito da irrigação do canal radicular por pressão apical negativa e por pressão positiva na expressão de moléculas que são indicativas de diferenciação celular com fenótipo mineralizador em dentes de cães com rizogênese incompleta e lesão periapical. Um total de 30 dentes (60 raízes) foi distribuído em 3 grupos (n=20): EndoVac, Convencional e Controle. Após 30 dias, foram realizados os procedimentos histotécnicos de rotina e os cortes foram submetidos à técnica de imunohistoquímica para marcação de Osteopontina (OPN), Fosfatase Alcalina (ALP) e para o fator de transcrição RUNX2 nas regiões apical e periapical das raízes. Foi realizada uma análise semi-quantitativa da imunomarcação positiva e a intensidade da expressão foi classificada em ausente (0), leve (1), moderada (2) ou intensa (3). Os dados por escores foram analisados estatisticamente pelo teste não-paramétrico de Kruskal-Wallis e pelo pós-teste de Dunn, e o nível de significância adotado foi de 5%. A imunumarcação para RUNX2 revelou que no grupo pessão negativa houve marcação significativamente mais intensa (p<0,05), em comparação ao grupo controle. Com relação à expressão de OPN, não foi possível observer diferença estatisticamente significante entre os grupos (p>0,05). Após análise da imunomarcação para ALP, foi observado diferença estatisticamente significante entre os grupos (p<0,05), e o grupo pressão negativa demonstrou uma marcação siginificativamente mais intensa do que o grupo controle. Os resultados do presente estudo in vivo permitiram concluir que a irrigação por pressão apical negativa apresenta potencial mineralizador em dentes com ápice aberto e lesão periapical.

Abstract The objective of this in vivo study was to assess the effect of the root canal irrigation by negative and positive apical pressure on the expression of molecules that are an indicative of cell differentiation with mineralizing phenotype in teeth of dogs with incomplete rhizogenesis and induced periapical lesion. A total of 30 teeth (60 roots) were distributed into 3 groups (n=20): EndoVac®, Conventional and Control. After 90 days, the routine histotechnical procedures were performed and the sections were submitted to immunohistochemical technique for the staining of osteopontin (OPN), alkaline phosphatase (ALP) and the RUNX2 transcription factor in the apical and periapical regions of the roots. A semi-quantitative analysis of the positive immunostaining was performed and the intensity of the expression was classified in absent (0), mild (1), moderate (2), or intense (3). Scores data were statistically analyzed by the Kruskal-Wallis non-parametric test and Dunn post-test, and the significance level was set at 5%. RUNX2 immunostaining revealed that in the negative pressure group there was a significantly stronger (p<0.05) immunostaining in comparison to the control group. Regarding the OPN expression, it was not possible to detect a statistically significant difference between the groups (p>0.05). After analyzing ALP immunostaining, a statistically significant difference was observed between the groups (p<0.05), and the negative pressure group showed a markedly stronger mark immunostaining than the control group. The results of the present in vivo study allowed concluding that negative apical pressure irrigation presents mineralizing potential in immature teeth with apical periodontitis.

Resumo O objetivo deste estudo foi avaliar a resposta subcutânea do tecido conjuntivo de camundongos isogênicos após o implante de diferentes cimentos à base de ionômero de vidro (EQUIA® Forte Fil, EQUIA® Fil e Ketac ™ Universal Aplicap ™). Oitenta e sete camundongos isogênicos BALB/c foram alocados em 12 grupos, 9 como grupos experimentais (Ketac, E. Fil e E. Forte aos 7, 21 e 63 dias) e 3 controles (tubos de polietileno vazios aos 7, 21 e 63 dias). Após os períodos experimentais, o tecido conjuntivo subcutâneo ao redor do material implantado foi removido e submetido ao processamento histotécnico e coloração com hematoxilina e eosina. Uma descrição histopatológica da reação tecidual envolvendo cada material e uma análise semi-quantitativa da fibrose e infiltrado inflamatório foram realizadas. Além disso, foi realizada a mensuração da espessura do tecido granulomatoso em contato com cada material. Os dados foram analisados estatisticamente (α=0,05) pelo teste de Kruskal-Wallis, seguido do pós-teste de Dunn. Inicialmente, a fibrose não foi diferente entre todos os materiais testados (p>0,05), mas foi diferente aos 21 dias, com o grupo controle apresentando o estágio mais avançado de fibrose. Aos 63 dias, o grupo EQUIA® Forte Fil apresentou o estágio mais avançado de fibrose, comparado ao grupo EQUIA® Fil (p<0,05). O infiltrado inflamatório não foi diferente entre os materiais testados em nenhum período experimental (p>0,05). A espessura do tecido granulomatoso foi maior no grupo E. Forte, comparado ao controle em todos os períodos. Todos os cimentos à base de ionômero de vidro apresentaram compatibilidade tecidual, de acordo com os parâmetros avaliados.

Abstract The aim of this study was to evaluate the subcutaneous connective tissue response of isogenic mice after implantation of different glass ionomer-based cements (EQUIA® Forte Fil, EQUIA® Fil and Ketac™ Universal Aplicap™). Eighty-seven isogenic BALB/c mice were allocated in 12 groups, 9 were considered as experimental groups (Ketac, E. Fil and E. Forte at 7, 21 and 63 days) and 3 controls (empty polyethylene tubes at 7, 21 and 63 days). After the experimental periods, the subcutaneous connective tissue surrounding the implanted material was removed and subjected to histotechnical processing and staining with hematoxylin and eosin. A histopathological description of the tissue reaction surrounding each material and a semi-quantitative analysis of collagen fiber formation and inflammatory infiltrate were performed. Additionally, the thickness of the granulomatous tissue in contact with each material was measured. Data were analyzed statistically (α=0.05) by the Kruskal-Wallis test, followed by Dunn post-test. Initially, the collagen fiber formation was not different among all the tested materials (p>0.05) but was different at 21 days with the control group presenting the most advanced stage of collagen fiber formation. At 63 days, EQUIA® Forte Fil group showed the most advanced stage of collagen fiber formation, compared to EQUIA® Fil group (p<0.05). The inflammatory infiltrate was not different among the tested materials in any experimental period (p>0.05). The thickness of the granulomatous tissue was greater in the E. Forte group, compared to control in all periods. All glass ionomer-based cements showed tissue compatibility, according to the evaluated parameters.

Abstract Objective To evaluate and correlate, in the same research, the mRNA expression and the staining of RANK, RANKL, OPG, TLR2 and MyD88 by immunohistochemistry in the apical periodontitis (AP) progression in mice. Material and Methods AP was induced in the lower first molars of thirty-five C57BL/6 mice. They were assigned to four groups according to their euthanasia periods (G0, G7, G21 and G42). The jaws were removed and subjected to histotechnical processing, immunohistochemistry and real-time reverse transcription-PCR (qRT-PCR). Data were analyzed with parametric and nonparametric tests (α=0.05). Results An increase of positive immunoreactivity for RANK, RANKL, OPG, TLR2 and MyD88 was observed over time (p<0.05). The RANKL expression was different between the groups G0 and G42, G21 and G42 (p=0.006), with G42 presenting the higher expression in both comparations. The OPG expression was statistically different between the groups G0 and G7, G7 and G21 and G7 and G42 (p<0.001), with G7 presenting higher expression in all the time points. The TLR2 expression was different between the groups G0 and G42 (p=0.03), with G42 showing the higher expression. The MyD88 expression presented a statistical significant difference between groups G7, G21 and G42 compared with G0 (p=0.01), with G0 presenting the smallest expression in all the comparisons. The Tnfrsf11/Tnfrsf11b (RANKL/OPG) ratio increased with the AP progression (p=0.002). A moderate positive correlation between MyD88 and RANKL (r=0.42; p=0.03) and between MyD88 and TLR2 (r=0.48; p<0.0001) was observed. Conclusion The expression of the RANK, RANKL, OPG, MyD88 and TLR2 proteins as well as the ratio Tnfrsf11/Tnfrsf11b (RANKL/OPG) increased with AP progression. There was also a moderate positive correlation between the expression Myd88-Tnfrsf11 and Tlr2-Myd88, suggesting the relevance of Tlr2-Myd88 in bone loss due to bacterial infection.

Abstract Aim: To compare the apical negative pressure irrigation (ANP) with conventional irrigation in the teeth of immature dogs with apical periodontitis. Methods: Fifty-two immature pre-molar root canals were randomly assigned into 4 groups: ANP (n=15); conventional irrigation (n=17); healthy teeth (control) (n = 10); and teeth with untreated apical periodontitis (control) (n=10). After induction of apical periodontitis, teeth were instrumented using EndoVac® (apical negative pressure irrigation) or conventional irrigation. The animals were euthanized after 90 days. The sections were stained by HE and analyzed under conventional and fluorescence microscopy. TRAP histoenzymology was also performed. Statistical analyses were performed with the significance level set at 5%. Results: There was difference in the histopathological parameters between ANP and conventional groups (p<0.05). The ANP group showed a predominance of low magnitude inflammatory infiltrate, a smaller periodontal ligament, and lower mineralized tissue resorption. There were no differences in the periapical lesion extensions between the ANP and conventional groups (p>0.05). However, a lower number of osteoclasts was observed in the ANP group (p<0.05). Conclusion: The EndoVac® irrigation system presented better biological results and more advanced repair process in immature teeth with apical periodontitis than the conventional irrigation system, confirming the hypothesis.

Resumo O objetivo do presente estudo foi avaliar propriedades físico-químicas da pasta à base de hidróxido de cálcio (Calen(r)) combinado com diferentes proporções de óxido de zinco (OZ) (1:0,5 e 1:0,65, 1:0,8 e 1:1) (Calen(r) /OZ). Os materiais foram comparados quanto ao tempo de endurecimento, variação do pH, radiopacidade, solubilidade, alterações dimensionais, escoamento e liberação de elementos químicos. Os dados obtidos foram submetidos à análise de variância (nível de 5% de significância). Maior grau de endurecimento, valores de alterações dimensionais, e solubilidade foram encontrados para Calen(r) /OZ 1:0,65 e 1:0,5. Calen(r) /OZ 1:0,5 e 1:0,65, apresentaram os maiores valores de pH em todos os momentos. Todos os materiais apresentaram valores elevados de radiopacidade com diferenças significativas (p>.05). Foram encontradas diferenças significativas apenas entre Calen(r) /OZ de 1:0,5 e 1:1 na liberação de cálcio e zinco (p< 0,05), enquanto que a quantidade de zircônia foi semelhante entre todos os grupos analisados (p>.05). Em relação às propriedades avaliadas no presente estudo, as combinações de Calen(r) com óxido de zinco nas proporções 1:0,5 e 1:0,65 são mais adequadas como material obturador para os canais radiculares de dentes decíduos.

Abstract This study evaluated physiochemical proprieties of a calcium hydroxide-based paste (Calen(r)) combined with a zinc oxide cement at different ratios (1:0.5, 1:0.65, 1:0.8 and 1:1). Materials were compared regarding setting time, pH variation, radiopacity, solubility, dimensional changes, flow and release of chemical elements. Data were analyzed statistically by ANOVA and Tukey's test (α=0.05). Longer setting time and higher dimensional changes and solubility values were exhibited by 1:0.65 and 1:0.5 ratios (p<0.05). The 1:0.5 and 1:0.65 ratios exhibited the highest pH values at all time points. All materials exhibited high radiopacity values. Significant differences were found only between 1:0.5 and 1:1 ratios for calcium and zinc release (p<0.05), whereas the amount of zirconium was similar among all groups (p>0.05). Considering the evaluated proprieties, combinations of Calen(r) paste with ZO at 1:0.5 and 1:0.65 ratios had the best results as root canal filling materials for use in primary teeth.