Apical Negative Pressure irrigation presents tissue compatibility in immature teeth

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). However, a lower number of osteoclasts was observed in the ANP group (

Minimally interventive restorative care of teeth with molar incisor hypomineralization and open apex—A 24‐month longitudinal study

Aim: To assess the efficacy of treatment using a minimally invasive approach (selective removal of carious tissue, restoration and preventive strategies) in immature permanent molars with MIH. Design: A total of 281 patients, aged 6-8 years, with carious lesions (ICDAS 5-6), severe MIH, and incomplete root formation (one tooth/patient) were included. After clinical and radiographic examinations, selective carious tissue removal was performed, and the teeth received interim restoration for 6 months and were then restored with composite resin. Clinical and radiographic follow-up was undertaken, 6, 12, 18, and 24 months. A protocol of preventive oral care measures was established and repeated at each follow-up, including diet counselling, oral hygiene instruction, dental plaque control, and topical application of fluoride varnish containing CPP-ACP. All clinical procedures and evaluations were done by a single operator. Results: Clinical and radiographic success was observed 24 months after treatment in 96.8% of the cases. Failures were due to enamel fracture at restoration margins, resulting in pulpitis and absence of apex closure. Conclusion: Selective removal of carious tissue, interim, and subsequently definitive restoration, combined with home and professional preventive measures, maintained marginal integrity of restorations in immature permanent molars with severe MIH, confirmed by pulp vitality and occurrence of apexogenesis

Immature teeth: comparative analysis of two types of root canals irrigation, in dogs\'s teeth with induced apical periodontitis

A lesão periapical é uma doença causada por micro-organismos presentes no sistema de canais radiculares, assim como por seus produtos e subprodutos e pelas reações desencadeadas no organismo. A base para a obtenção do sucesso pós-tratamento endodôntico é o controle da infecção. Com essa finalidade, novos sistemas de irrigação dos canais radiculares têm sido desenvolvidos, no qual a irrigação por Pressão Apical Negativa (EndoVac®) tem se destacado. O objetivo desse estudo in vivo foi efetuar uma avaliação histopatológica, histoenzimológica e histomicrobiológica, comparando o sistema EndoVac® com a irrigação convencional, em dentes de cães com rizogênese incompleta e lesão periapical crônica induzida experimentalmente. Um total de 52 canais radiculares de pré-molares com rizogênese incompleta foram aleatoriamente divididos em 4 grupos: Grupo I - Irrigação com EndoVac® (n=15); Grupo II - Irrigação Convencional (n=17); Grupo III - Dente hígido (controle negativo) (n=10) e Grupo IV - Dente com lesão e sem tratamento (controle positivo) (n=10). Após indução de lesões periapicais nos dentes dos Grupos I, II e IV, os dentes dos grupos I e II foram instrumentados com limas manuais, empregando o EndoVac® e a irrigação convencional, respectivamente. Decorridos 90 dias, os animais foram eutanasiados e os espécimes submetidos ao processamento histotécnico para a análise histopatológica morfológica e morfométrica em cortes corados com HE, sob microscopia convencional e de fluorescência. Foi realizada também a histoenzimologia para a Fosfatase Ácida Resistente ao Tartarato (TRAP), para a marcação de osteoclastos, e a coloração de Brown e Brenn, para avaliar a presença de bactérias, sua localização e concentração/intensidade. Os resultados obtidos foram submetidos à análise estatística por meio dos testes de qui-quadrado, Fisher, Anova e pós-teste de Tukey. O nível de significância adotado foi de 5%. Os resultados obtidos na avaliação dos parâmetros histopatológicos evidenciaram diferença significante entre os Grupos I e II (p<0,05), observando-se no Grupo I predomínio de infiltrado inflamatório de menor magnitude, menor espessura do ligamento periodontal e menor reabsorção dos tecidos mineralizados. Embora não tenha sido verificada diferença significante entre esses grupos com relação à extensão das lesões periapicais em microscopia de fluorescência, e com relação à presença, localização e intensidade da contaminação bacteriana, foi observado menor número de osteoclastos no Grupo I (p<0,05). Os resultados do presente estudo in vivo permitiram concluir que a irrigação por pressão apical negativa (EndoVac®) apresentou melhores resultados biológicos, quando comparada à irrigação convencional, favorecendo a ocorrência do processo de reparo, em dentes com rizogênese incompleta e lesão periapical.The apical periodontitis is a disease caused by microorganisms from the root canal system, as well as their products, by-products and the reactions triggered in the host. The postendodontic treatment success is based in the control of the infection. For this purpose, new irrigation systems of root canals have been developed, in which apical Negative Pressure (EndoVac®) have been highlighted. The objective of this in vivo study was to perform a histopathological, histoenzymology and histomicrobiologic evaluation, comparing the EndoVac® system with the conventional irrigation, in immature teeth of dogs with apical periodontitis experimentally induced. A total of 52 root canals of immature pre-molars were randomly assigned into 4 groups: Group I - EndoVac® Irrigation (n= 15); Group II - Conventional Irrigation (n= 17); Group III - Sound Tooth (negative control) (n= 10) and Group IV - Tooth with apical periodontitis without treatment (positive control) (n= 10). After induction of apical periodontitis in teeth of the Groups I, II and IV, in the groups I and II, the teeth were instrumented with manuals files, using EndoVac® and conventional irrigation, respectively. After 90 days, the animals were euthanized and the specimens subjected to histotechnic for histopathological, morphologic and morphometric analysis in HE-stained sections, under conventional and fluorescence microscopy. The histoenzymology for the phosphatase-Resistant Acid tartrate (TRAP), for the identification of osteoclasts, and the Brown and Brenn staining method to assess the presence of bacteria, their location and concentration/intensity were also performed. The results obtained were subjected to statistical analysis using the chi-square test, Fisher exact test, ANOVA and Tukey post-test. The level of significance was set at 5%. The results obtained in the evaluation of histopathological parameters demonstrated a significant difference between groups I and II (p< 0.05). In the Group I, the predominance was inflammatory infiltrate of low magnitude, small thickness of the periodontal ligament and less resorption of mineralized tissues. Although the periapical lesions extension in fluorescence microscopy, and the presence, location, and intensity of bacterial contamination was not significant different between these groups, a lower number of osteoclasts in Group I (p< 0.05) was observed. The results of this in vivo study allowed us to conclude that the irrigation with EndoVac® presents best biological results in relation to conventional irrigation, favored the occurrence of the repair process, in immature teeth with apical periodontitis

Cells and molecules characterization of the innate immune system involved in the progression and treatment of apical periodontitis and the development of periodontal disease

O sistema imune inato representa a primeira de linha de defesa do nosso organismo. Receptores de Reconhecimento de Padrões (PRRs) são expressos pelas células desse sistema, como os macrófagos, reconhecendo os micro-organismos e apresentando os antígenos ao sistema imune adaptativo. Dentre os PRRs destaca-se o IFI16 (Proteína induzível por interferon gama 16), uma proteína multifuncional com ação anti-inflamatória. No entanto, o papel da IFI16 na Odontologia ainda é pouco conhecido. Assim, o primeiro o objetivo desse trabalho foi caracterizar a fonte celular da IFI16 na progressão da periodontite induzida experimentalmente em camundongos, a fim de verificar se as células derivadas da medula óssea poderiam ser a fonte da expressão da IFI16 na periodontitite. Inicialmente foi realizada a depleção da medula óssea de animais wild-type (WT) e de animais Ifi204-/- (ortólogo para IFI16). Em seguida, foi realizado o transplante de medula óssea entre animais Ifi204-/- e WT. Após 21 dias do transplante, a periodontite foi induzida em ambos os grupos experimentais pelo modelo de ligadura. Decorridos 9 dias, foi realizada citometria de fluxo da medula óssea utilizando o CD45 como um marcador hematopoiético para determinar o sucesso do transplante. A perda óssea alveolar foi medida pela análise de µCT. Os dados foram analisados pelo teste \"t\" usando o software GraphPad Prism 7.0a (a=5%). Trinta dias após o transplante, a eficiência de reconstituição encontrada foi de cerca de 80% com base no CD45.1/2. Camundongos com as células transplantadas do animal Ifi204-/-, não mostraram diferença significativa de perda óssea média em comparação com camundongos com as células transferidas de animal WT [0,35mm (DP=0,058mm) vs. 0,43mm (DP=0,104mm) respectivamente, p>0,05]. Assim, o transplante de medula óssea demonstrou que a origem das células que expressam IFI16 durante a progressão da periodontite não são de origem hematopoiética. O segundo objetivo desse estudo foi avaliar, por meio de imunohistoquímica, a participação da IFI16 e IFN-&alpha;/&beta; na gênese e no desenvolvimento da lesão periapical induzida em dentes de camundongos. De um total de 45 camundongos C57BL/6, foi realizada a indução da lesão periapical nos primeiros molares inferiores em 40 animais divididos nos períodos experimentais de 2, 7, 14, 21 e 42 dias. Oito animais de cada período experimental e 5 animais do grupo controle foram eutanasiados e os espécimes submetidos ao processamento histotécnico para imunomarcação para a descrição da presença/ausência e localização da IFI16 e IFN-&alpha;/&beta; (análise qualitativa). Após análise dos resultados, concluiu-se que a IFI16 e os IFN-&alpha;/&beta; participaram da progressão da lesão periapical induzida experimentalmente em dentes de camundongos. Paralelamente, os macrófagos são células fagocíticas, conhecidas por apresentarem uma considerável heterogeneidade em termos morfológicos, funcionais e expressão de marcadores. Dependendo do microambiente ao qual os monócitos, suas células progenitoras, estão expostos, há a definição do seu fenótipo em \"classicamente ativado\" (M1) ou \"alternativamente ativado\" (M2). Os macrófagos M1 promovem um ambiente altamente microbicida e têm papel na mediação da destruição de patógenos e células tumorais. Já os macrófagos M2 desempenham um papel central nas respostas frente às infestações parasitárias, remodelação tecidual, angiogênese e doenças alérgicas. Assim, o terceiro objetivo desse trabalho foi a caracterização fenotípica de macrófagos M1 e M2 no desenvolvimento da lesão periapical induzida em dentes de camundongos. Um total de 130 camundongos C57BL/6 foram divididos em grupos controle (dentes hígidos; n=50 animais) e experimental (dentes com lesão periapical experimentalmente induzida; n=80 animais). Decorridos os períodos de 2, 7, 14, 21 e 42 dias, animais controles e experimentais foram eutanasiados e os espécimes submetidos ao processamento histotécnico, para a descrição das características dos tecidos apicais e periapicais em cortes corados com HE, sob microscopia convencional. Além dissa, sob microscopia de fluorescência foi realizada a morfometria para medir a área das lesões periapicais. Foi realizado, também, o qRT-PCR para análise de expressão gênica de Cxcl10, CxCL9, iNos2, Arg1, Ym1, Fizz1 e MRC1 e ensaio Luminex®, para a marcadores de macrófagos M1 e M2 (GM-CSF, IFN-g, IL-4, IL-13, IL- 10, IL-6, IL-1&beta; e TNF-&alpha;). Para os dados da morfometria realizou-se o teste ANOVA, seguido pelo pósteste de Tukey. Para os valores obtidos de qRT-PCR e Luminex®, realizou-se o teste de Kruskal-Wallis seguido do pós-teste de Dunn. Todos os dados foram analisados usando o software GraphPad Prism 7.0a (a=5%). Os resultados permitiram concluir que no decorrer dos períodos, a lesão periapical progrediu. Além disso, com base nos resultados de expressão gênica e quantificação das citocinas, observou-se que a progressão da lesão periapical ocorreu de maneira dinâmica. Além disso, houve predomínio do fenótipo M1 nos períodos iniciais da lesão periapical, passando pela tentativa de reparo com o fenótipo M2 aos 14 e 21 dias e voltando ao fenótipo M1 quando a lesão está estabelecida e é caracterizada como crônica. Por fim, o quarto objetivo desse estudo foi avaliar a capacidade de diferentes materiais utilizados no tratamento endodôntico de dentes portadores de necrose pulpar e lesão periapical em modular o fenótipo de macrófagos M1 e M2. Em cultura de células de macrófagos derivados da medula óssea de animais, foi realizada a análise da expressão gênica de marcadores de macrófagos M1 e M2 por meio do qRT-PCR (Cxcl10, CxCL9, iNos2, Arg1 , Ym1, Fizz1 e MRC1) e quantificação de citocinas pelo Luminex® (GM-CSF, IFN-&gamma;, IL-4, IL-13, IL- 10, IL-6, IL-1&beta; e TNF-&alpha;), após exposição aos cinco cimentos endodônticos AH PlusTM, Sealapex XpressTM, EndosequenceTM, BioRootTM, EndomethasoneTM e a pasta Calen®. Para os valores normais utilizou-se ANOVA, seguido pelo pós-teste de Tukey. Para os valores não normais, utilizou-se o teste de Kruskall-Wallis. Todos os dados foram analisados usando o software GraphPad Prism 7.0a (&alpha;=5%). Conclui-se que os materiais avaliados foram capazes de induzir a produção de citocinas pró e anti-inflamatórias, além de estimularem um padrão misto de macrófagos (M1/M2).The innate immune system represents the first defense line of our body. Pattern Recognition Receptors (PRR) are expressed by cells in this system, such as macrophages. They recognize microorganisms and present the antigens to the adaptive immune system. Among the PRRs, IFI16 (interferon gamma inducible protein 16) stands out. It is a multifunctional protein with anti-inflammatory function. However, the role of IFI16 in dentistry is still poorly understood. Thus, the first aim of this study was to characterize the cellular source of IFI16 in the progression of periodontitis experimentally induced in mice. In order to verify if bone marrow-derived cells were the source of IFI16 expression in the periodontitis, was initially performed the bone marrow depletion of animals wild-type (WT) and Ifi204-/- (ortolog to IFI16). Then, bone marrow transplant (BMT) was performed between Ifi204-/- and WT animals. After 21 days of BMT, periodontitis was induced in both groups using the ligature model. In order to determine BMT success, after 9 days was performed flow cytometry using cells from bone marrow. CD45 was used as a hematopoietic marker. Alveolar bone loss was measured by µCT analysis. Data were analyzed using t-test with GraphPad Prism 7.0a software (&alpha;=5%). Thirty days after BMT, based on CD45.1/2, the reconstitution efficiency was about 80%. Mice with transplanted cells from animal Ifi204-/- showed no significant difference in mean bone loss compared to mice with animal transferred cells from WT [0.35mm (SD=0.058mm) vs. 0.43mm (SD=0.104mm), respectively p>0.05]. Thus, BMT experiment demonstrated that the source of the cells expressing IFI16 during the progression of periodontitis are not from hematopoietic origin. The second aim of this study was to evaluate, by immunohistochemistry, the participation of IFI16 and IFN-&alpha;/&beta; in the genesis and development of apical periodontitis induced in mice teeth. From a total of 45 mice C57BL/6, in 40 animals the apical periodontitis was induced in the lower first molars. They were divided in experimental periods of 2, 7, 14, 21 and 42 days. Eight animals from each experimental period and 5 animals from the control group were euthanized and the specimens submitted to histological process and submitted to immunostaining. After was performed the description of the presence/absence and localization of IFI16 and IFN-&alpha;/&beta; (qualitative analysis). After the results analysis, it was concluded that IFI16 and IFN-&alpha;/&beta; participated in the progression of apical periodontitis experimentally induced mice teeth. At the same time, macrophages are phagocytic cells, known to have considerable morphological, functional and marker expression heterogeneity. Depending on the microenvironment to which the monocytes, their progenitor cells, are exposed, their phenotype is defined as \"classically activated\" (M1) or \"alternatively activated\" (M2). M1 macrophages promote a highly microbicidal environment and play a role in mediating the destruction of pathogens and tumor cells. M2 macrophages play a central role in responses to parasitic infestations, tissue remodeling, angiogenesis and allergic diseases.Thus, the third aim of this work was the phenotypic characterization of M1 and M2 macrophages in the development of apical periodontitis induced in mice teeth. A total of 130 mice C57BL/6 were separated into control (healthy teeth; n=50 animals) and experimental (teeth with experimentally induced apical periodontitis; n=80 animals) groups. After the periods of 2, 7, 14, 21 and 42 days, control and experimental animals were euthanized and the specimens submitted to histotechnical process in order to describe the apical and periapical tissue characteristics in HE stained sections under conventional microscopy. Besides that, under fluorescence microscopy, morphometry was performed to measure the area of the apical periodontitis. Also, were performed qRT-PCR for Cxcl10, CxCL9, iNos2, Arg1, Ym1, Fizz1 and MRC1 gene expression analysis and Luminex® assay for M1 and M2 macrophage markers (GM-CSF, IFN-g, IL-4, IL-13, IL-10, IL-6, IL-1&beta; and TNF-&alpha;). For morphometry data, the ANOVA test was performed, followed by Tukey post-test. For the values obtained for qRT-PCR and Luminex®, the Kruskal-Wallis test was performed followed by the Dunn post-test. All data were analyzed using GraphPad Prism 7.0a software (a=5%). The results allowed to conclude that during the experimental periods, the apical periodontitis progressed. In addition, based on the results of gene expression and cytokine quantification, it was observed that the progression of the apical periodontitis occurred dynamically. Besides that, there was a predominance of the M1 phenotype in the early periods of the apical periodontitis, attempting to repair the M2 phenotype at 14 and 21 days and returning to the M1 phenotype when the lesion is established and characterized as chronic process. Finally, the fourth aim of this study was to evaluate the ability of different materials used during endodontic treatment of teeth with pulp necrosis and apical periodontitis in modulate the M1 and M2 macrophage phenotype. In bone marrow-derived macrophage cell culture, gene expression analysis of markers to M1 and M2 macrophages was performed by qRT-PCR (Cxcl10, CxCL9, iNos2, Arg1, Ym1, Fizz1 and MRC1) and cytokine quantification by Luminex® (GM-CSF, IFN-g, IL-4, IL-13, IL-10, IL-6, IL-1&beta; and TNF-&alpha;) after exposure to the five endodontic sealers AH PlusTM Sealapex XpressTM, EndosequenceTM, BioRootTM, EndomethasoneTM; and the Calen® paste. For normal values, ANOVA test was used, followed by Tukey post-test. For non-normal values, the Kruskall-Wallis test was used. All data were analyzed using GraphPad Prism 7.0a software (a=5%). It was concluded that the evaluated materials were able to induce the expression of pro and anti-inflammatory cytokines, besides stimulating a mixed macrophage pattern (M1/M2)

Color Stability and Surface Roughness of Composites after Artificial Accelerated Aging

<p>The color stability and surface uniformity are very important properties for dental aesthetics. <strong>Objective: </strong>To evaluate the color stability and surface roughness of different composites after artificial accelerated aging (AAA). <strong>Methods: </strong>Samples were made using the silorane-based Filtek P90 (3M-ESPE), nanohybrid Tetric N-Ceram (Ivoclar Vivadent), and GC Kalore (GC Company). Ceramic D. Sign (Ivoclar Vivadent) and Ketac N100 (3M-ESPE) resin-modified glass ionomer cement (RMGIC) served as controls. The values for color stability and surface roughness were recorded before and after AAA for non-C-UV (300 hours). Color stability was assessed as the difference between the coordinates obtained from the L*a*b* system. The surface roughness was analyzed with a rugosimeter. The surface value of each sample was taken as the average of these measurements. The one-way ANOVA and Tukey’s post-test with α=5% was used. <strong>Results: </strong>The greatest change in color stability occurred for the RMGIC (ΔE=18.7) and the least for ceramics (ΔE=2.1). No significant difference was noted among the composites (p&gt;0.05). The surface roughness before and after AAA differed significantly only for the RMGIC (p&lt;0.05). <strong>Conclusion: </strong>The two latest generation resins (Filtek P90 and GC Kalore) showed good results in terms of color stability and surface roughness for use in aesthetic restorations.</p><p> </p

Apical Negative Pressure irrigation presents tissue compatibility in immature teeth

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 (p0.05). 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

Immunohistochemical and mRNA expression of RANK, RANKL, OPG, TLR2 and MyD88 during apical periodontitis progression in mice

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