Abstract High expression of MMP-2 and MMP-9 in periapical lesions plays an important role in the degradation of the extracellular matrix. This study aimed to investigate the effect of epigallocatechin-3-gallate (EGCG)-based endodontic paste as an intracanal dressing on the expression of MMP-2 and MMP-9 in periapical lesions. Periapical lesions were experimentally induced in 35 mature beagle dog premolars randomly divided into healthy teeth, untreated periapical lesions, periapical lesions treated in a single session (control groups), and periapical lesions treated in two sessions with EGCG or calcium hydroxide-based pastes (experimental groups). After 120 days, specimens were obtained for histopathologic and immunofluorescence analyses to assess the expression of MMP-2 and MMP-9. The statistical analysis was performed using a p-value of 0.05. Endodontic treatment in two sessions using medication with EGCG and calcium hydroxide-based pastes provided similar repair of the apical and periapical tissues and neoformation of periodontal ligament fibers, cementum, and alveolar bone (p>0.05). The experimental groups treated in two sessions with both medications presented expression of MMP-2 and MMP-9 similar to that in healthy teeth (p>0.05), and significantly lower than teeth treated in a single session or untreated periapical lesions (p <0.001). Expression of MMP-2 and MMP-9 was observed in the cytoplasm of fibroblasts, osteoblasts, cementoblasts, cementocytes, and vascular endothelium. The use of EGCG-based endodontic paste reduced the expression of MMP-2 and MMP-9 and allowed repair of periapical lesions, similar to calcium hydroxide-based paste, and superior to treatment performed in a single session. MMP2 MMP 2 MMP- MMP9 9 matrix epigallocatechin3gallate epigallocatechingallate epigallocatechin 3 gallate EGCGbased based control groups, , groups) hydroxidebased hydroxide groups. . 12 days MMP9. 9. pvalue p value 005 0 05 0.05 fibers cementum p>0.05. p005 p>0.05 (p>0.05) p>0.05, <0.001. 0001 <0.001 001 <0.001) fibroblasts osteoblasts cementoblasts cementocytes endothelium 1 00 0.0 p00 p>0.0 (p>0.05 000 <0.00 0. p0 p>0. (p>0.0 <0.0 p>0 (p>0. <0. p> (p>0 <0 (p> <

Resumo A alta expressão de MMP-2 e MMP-9 em lesões periapicais desempenha um papel importante na degradação da matriz extracelular. Este estudo teve como objetivo investigar o efeito de uma pasta à base de epigalocatequina-3-galato (EGCG) como curativo intracanal na expressão de MMP-2 e MMP-9 em lesões periapicais. Lesões periapicais foram induzidas experimentalmente em 35 pré-molares de cães da raça beagle, maduros, divididos aleatoriamente em dentes saudáveis, lesões periapicais não tratadas, lesões periapicais tratadas em uma única sessão e lesões periapicais tratadas em duas sessões com a pasta de EGCG ou pasta à base de hidróxido de cálcio. O operador monitorou os animais e realizou a eutanásia após 120 dias para análises histopatológicas e de imunofluorescência para avaliar a expressão de MMP-2 e MMP-9. A análise estatística foi realizada utilizando p=0,05. O tratamento endodôntico em duas sessões com pasta à base de EGCG e pasta à base de hidróxido de cálcio proporcionou níveis semelhantes de reparação dos tecidos apicais e periapicais e neoformação de fibras do ligamento periodontal, cemento e osso alveolar. Em ambos os grupos, a expressão de MMP-2 e MMP-9 foi mínima, sendo observada no citoplasma de fibroblastos, osteoblastos, cementoblastos, cementócitos e endotélio vascular. Em ambos os grupos tratados em duas sessões, a expressão de MMP-2 e MMP-9 foi semelhante à dos dentes hígidos e significativamente menor do que nas lesões periapicais tratadas em sessão única ou não tratadas (p < 0,001). O uso da pasta à base de EGCG reduziu a expressão de MMP-2 e MMP-9 e permitiu o reparo de lesões periapicais, semelhante à pasta à base de hidróxido de cálcio, e foi superior ao tratamento realizado em sessão única. MMP2 MMP 2 MMP- MMP9 9 extracelular epigalocatequina3galato epigalocatequinagalato epigalocatequina 3 galato (EGCG prémolares pré molares beagle maduros saudáveis 12 MMP9. 9. p005 p 0 05 p=0,05 periodontal alveolar mínima fibroblastos osteoblastos cementoblastos vascular 0,001. 0001 0,001 . 001 0,001) 1 p00 p=0,0 000 0,00 00 p0 p=0, 0,0 p=0 0, p=

Abstract Brazil’s public healthcare system (SUS) offers specialized oral health services to Brazilians, but the productivity of specialists, such as Pediatric Dentists, has not been characterized. Therefore, the objective of this study was to characterize the outpatient dental procedures (ODPs) carried out by Pediatric Dentists within the SUS. An epidemiological study with an ecological, longitudinal, retrospective, and quantitative approach was conducted. The ODPs carried out by Pediatric Dentists within the SUS were characterized based on type of procedure, complexity level, and circumstance (urgent or elective). Data were analyzed using a descriptive and analytical approach, considering a significance level of 5%, as well as the impact of the COVID-19 pandemic (the 2020-2022 years were not included in secondary analyses). In the last 15 years, 29,234,972 ODPs were carried out by Pediatric Dentists within the SUS. Clinical procedures were the majority (55.4%), significantly more frequent than all other types of procedures (all p <0.05). Among these, restorative and periodontal procedures were the most common (30.7% and 21.0%, respectively). From 2008 to 2019, excluding COVID-19 pandemic years, the trend over the last 15 years was constant for all types of procedures (all p >0.05). In addition, low complexity ODPs were the majority (90.1%), significantly more frequent than medium (9.7%) and high complexity procedures (0.1%) (both p <0.05). At last, most ODPs were not characterized by circumstance in the outpatient production reports (96.9%). Therefore, it was possible to conclude that Pediatric Dentists carried out diverse ODPs within the SUS over the past 15 years, although there was a dominant pattern of type and complexity. Brazils Brazil s (SUS Brazilians specialists Therefore (ODPs ecological longitudinal retrospective conducted procedure urgent elective. elective . elective) 5 5% COVID19 COVID 19 COVID-1 20202022 2020 2022 2020-202 analyses. analyses analyses) 1 29234972 29 234 972 29,234,97 55.4%, 554 55.4% , 55 4 (55.4%) <0.05. 005 <0.05 0 05 <0.05) these 30.7% 307 30 7 (30.7 210 21 21.0% respectively. respectively respectively) 200 2019 >0.05. >0.05 >0.05) addition 90.1%, 901 90.1% 90 (90.1%) 9.7% 97 9 (9.7% 0.1% 01 (0.1% both 96.9%. 969 96.9% 96 (96.9%) COVID1 COVID- 2020202 202 2020-20 2923497 2 23 29,234,9 55.4 (55.4% 00 <0.0 30.7 3 (30. 21.0 20 201 >0.0 90.1 (90.1% 9.7 (9.7 0.1 (0.1 96.9 (96.9% 202020 2020-2 292349 29,234, 55. (55.4 <0. 30. (30 21. >0. 90. (90.1 9. (9. 0. (0. 96. (96.9 20202 2020- 29234 29,234 (55. <0 (3 >0 (90. (9 (0 (96. 2923 29,23 (55 < ( > (90 (96 292 29,2 (5 29,

Abstract Orthodontics patients usual develop demineralization and present cavity caries lesions after six months. Minimally invasive procedures have been the goal in modern dental practice. The aim of this study was to evaluate the effect of ClinproTMXT Varnish, on the enamel surface roughness and severity of white spot lesions. Twenty premolars were submitted to bond brackets and experimental induction of demineralization and randomly divided into 2 groups: GI - fluoride varnish (Colgate Duraphat®); GII - Ionomeric Sealant (ClinproTMXT Varnish). The treatment was applied around the brackets. The surface roughness of specimens was analyzed, before treatment and 12 weeks after treatment by laser confocal microscopy, and the severity of the white spot lesion was by laser fluorescence device. The data were analyzed by non-parametric Wilcoxon and Mann-Whitney Test, at 5% significance, roughness percentage reduction was performed. The severity of demineralization decreased in both, GI (p = 0.005) and GII (p = 0.019). Enamel superficial roughness levels decreased in GI and GII. As well as the roughness percentage, being more expressive in the ClinproTMXT Varnish group (85,09%). Colgate Duraphat® or Clinpro™ XT Varnish reduced the severity of the demineralization and decreased the superficial roughness on the enamel. The Clinpro™ XT Varnish was superior to superficial roughness on enamel. months practice groups Duraphat Duraphat®) Varnish. . Varnish) 1 microscopy device nonparametric non parametric MannWhitney Mann Whitney Test 5 significance performed both p 0.005 0005 0 005 0.019. 0019 0.019 019 0.019) 85,09%. 8509 85,09% 85 09 (85,09%) Clinpro 0.00 000 00 001 0.01 01 850 85,09 8 (85,09% 0.0 85,0 (85,09 0. 85, (85,0 (85, (85 (8 (

Resumo Pacientes ortodônticos geralmente desenvolvem desmineralização e apresentam lesões de cárie após seis meses de tratamento. Procedimentos minimamente invasivos têm sido o objetivo na prática odontológica moderna. O objetivo deste estudo foi avaliar o efeito do ClinproTM XT selante ionomérico, ao redor do bráquete, com relação a rugosidade superficial do esmalte e a severidade da lesão induzida. Vinte pré-molares foram submetidos a colagem de bráquetes e indução experimental de desmineralização e divididos aleatoriamente em 2 grupos: GI - verniz fluoretado (Colgate Duraphat®); GII - Selante Ionomérico (ClinproTM XT). O tratamento foi aplicado ao redor dos bráquetes. A rugosidade da superfície dos espécimes foi analisada, antes do tratamento e 12 semanas após o tratamento por microscopia confocal a laser e severidade da lesão de mancha branca por dispositivo de fluorescência a laser. Os dados foram analisados pelo teste não paramétrico de Wilcoxon e Mann-Whitney, a 5% de significância. A taxa de redução da lesão foi calculada. A severidade da desmineralização diminuiu tanto no GI (p = 0,005) quanto no GII (p = 0,019). Os níveis de rugosidade superficial do esmalte diminuíram no GI e GII, assim como o percentual de rugosidade, sendo mais expressivo no grupo ClinproTMXT (85,09%). Colgate Duraphat® e Clinpro™ XT reduziram a severidade da desmineralização e diminuíram a rugosidade superficial do esmalte. O selante ionomérico Clinpro™ XT foi superior na redução percentual de rugosidade. moderna bráquete induzida prémolares pré molares grupos Duraphat Duraphat®) XT. . XT) analisada 1 MannWhitney, MannWhitney Mann Whitney, Whitney Mann-Whitney 5 significância calculada p 0,005 0005 0 005 0,019. 0019 0,019 019 0,019) 85,09%. 8509 85,09% 85 09 (85,09%) Clinpro 0,00 000 00 001 0,01 01 850 85,09 8 (85,09% 0,0 85,0 (85,09 0, 85, (85,0 (85, (85 (8 (

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. Objective universitybased university based study Methods Brazil lesions 201 2018 recorded Results 5236 5 236 5,23 69 31 18% 18 (18% 1%. 1 1% . (1%) Conclusion followup follow up observed 20 523 23 5,2 3 (18 (1% 2 52 5, (1 (

Abstract This study aimed to evaluate the effect of Bifidobacterium animalis subsp. lactis (B. lactis) HN019 in drinking water on the development of apical periodontitis (AP) in rats. In total 60 animals were divided into a control group (sound teeth); Group I - regular water without AP; Group II - probiotic water without AP; Group III - regular water with AP; Group IV - probiotic water with AP. AP was induced after 3 days in the control groups and after 7, 21, and 42 days in groups III and IV. The animals were euthanized, and the mandibles were subjected to histotechnical processing. Samples were stained with hematoxylin & eosin (H&E) to identify root canal features, apical and periapical regions. Additionally, histoenzymology was performed to detect osteoclasts, immunohistochemistry was used to identify osteoclastogenesis markers, and the Brown & Brenn technique was applied for microbiological analysis. The data were analyzed using GraphPad Prism 8.0.1 with a significance level of 5%. Although no statistical differences were observed, the groups administered with probiotics showed better conditions in terms of histological aspects seen microscopically. Furthermore, there were no differences in the number of osteoclasts (p > 0.05). The RANKL marker was not found in the probiotic group at 42 days, unlike in group III. subsp B. B (B HN HN01 (AP rats 6 sound teeth teeth) 7 21 4 euthanized processing H&E HE H E (H&E features regions Additionally markers analysis 801 8 0 1 8.0. 5 5% observed microscopically Furthermore p 0.05. 005 0.05 . 05 0.05) HN0 2 80 8.0 00 0.0 8. 0.

Abstract Dental caries is an important Public Health issue. However, the treatment of this disease in tertiary dental care requires further investigation. Therefore, the objective of this study was to evaluate caries-related hospital morbidity in the Brazilian Unified Health System (SUS) from 2008 to 2022. An ecological study was conducted with secondary data on caries-related Hospital Admission Authorizations (AIH) and in-hospital dental procedures (IDP). Data were collected nationwide and statistical analysis was performed with a significance level (p) of 5%. In the last 15 years, 3,474 caries-related AIH and 63,657 IDP were approved within SUS. There was a significant upward trend in the number of caries-related AIH (p = 0.018) and a stationary trend in the number of caries-related IDP (p = 0.841). Moreover, from 2008 to 2022, R$ 1,160,843.09 was allocated for caries-related AIH. Hospital mortality was 0.29% (10 deaths), and 75.1% were elective inpatient admissions. Among SUS users, adults constituted the majority (49%), with a higher frequency of males (59.8%) and whites (46.2%). The most frequent type of caries-related IDP was restoration of permanent teeth (55.3%). Nonetheless, during the COVID-19 pandemic years, significant reductions in caries-related AIH and IDP within the SUS were observed (all p <0.05). Thus, within the SUS, caries-related tertiary dental care has shown a specific pattern over the last 15 years, including an increase in hospitalizations and a high number of in-hospital dental procedures, especially before the COVID-19 pandemic onset. issue However investigation Therefore cariesrelated related (SUS 200 2022 (AIH inhospital IDP. . (IDP) 5 5% 1 years 3474 3 474 3,47 63657 63 657 63,65 0.018 0018 0 018 0.841. 0841 0.841 841 0.841) Moreover R 116084309 160 843 09 1,160,843.0 029 29 0.29 10 (1 deaths, deaths , deaths) 751 75 75.1 admissions users 49%, 49 49% (49%) 59.8% 598 59 8 (59.8% 46.2%. 462 46.2% 46 2 (46.2%) 55.3%. 553 55.3% 55 (55.3%) Nonetheless COVID19 COVID 19 COVID-1 all <0.05. 005 <0.05 05 <0.05) Thus onset 20 202 (IDP 347 47 3,4 6365 6 65 63,6 0.01 001 01 084 0.84 84 11608430 16 1,160,843. 02 0.2 ( 7 75. 4 (49% 59.8 (59.8 46.2 (46.2% 55.3 (55.3% COVID1 COVID- 00 <0.0 34 3, 636 63, 0.0 08 0.8 1160843 1,160,843 0. (49 59. (59. 46. (46.2 55. (55.3 <0. 116084 1,160,84 (4 (59 (46. (55. <0 11608 1,160,8 (5 (46 (55 < 1160 1,160, 116 1,160 11 1,16 1,1 1,

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. Objective verify evaluations dentinpulpal dentin pulpal Biodentine followup follow up 3 6 1 Methods n 15 age sampling tissue lightcured light cured restoration Fishers Fisher s utilized Results However p<0.0001. p00001 p p<0.0001 . 0 0001 (p<0.0001) p=0.0013. p00013 p=0.0013 0013 (p=0.0013) Radiographically lesions 13 Biodentine™treated Biodentinetreated treated p<0.0013. p<0.0013 (p<0.0013) Conclusion treatment discoloration p0000 p<0.000 000 (p<0.0001 p0001 p=0.001 001 (p=0.0013 p<0.001 (p<0.0013 p000 p<0.00 00 (p<0.000 p=0.00 (p=0.001 (p<0.001 p00 p<0.0 (p<0.00 p=0.0 (p=0.00 p0 p<0. (p<0.0 p=0. (p=0.0 p<0 (p<0. p=0 (p=0. p< (p<0 p= (p=0 (p< (p= (p

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.

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 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.

Resumo Este estudo avaliou a associação entre polimorfismos em genes que codificam os receptores de estrogênio 1 (ESR1) e 2 (ESR2), receptor de vitamina D (VDR) e no microRNA17 (que se liga à ESR1 e VDR) e a periodontite apical persistente (PAP) após o tratamento endodôntico. Foram incluídos 162 pacientes com tratamento endodôntico concluído há pelo menos um ano e que apresentavam periodontite apical no início da terapia endodôntica. Exames clínicos e radiográficos foram realizados para avaliar a presença de PAP ou tecidos perirradiculares saudáveis (cicatrizados). As amostras de saliva foram coletadas como fonte de DNA genômico. A genotipagem de ESR1 (rs2234693 e rs9340799), ESR2 (rs1256049 e rs4986938), VDR (rs739837 e rs2228570) e miRNA17 (rs4284505) foram realizadas por PCR em tempo real. O teste do qui-quadrado foi utilizado para a distribuição das frequências genotípicas e alélicas. A análise de haplótipos também foi realizada. Oitenta e nove pacientes foram incluídos no grupo “curado” e 73 no grupo “PAP”. Não foi encontrada associação entre os polimorfismos alélicos e genotípicos estudados e a PAP (p>0,05). Concluí-se que os polimorfismos genéticos em ESR1, ESR2, VDR e miRNA17 não estão associados à PAP.

Abstract This study evaluated the association between polymorphisms in genes encoding estrogen receptors 1 (ESR1) and 2 (ESR2), vitamin D receptor (VDR) and in microRNA17 (which binds to ESR1 and VDR) with persistent apical periodontitis (PAP) after the endodontic treatment. We included 162 patients who completed endodontic treatment at least one year ago and presented apical periodontitis at the beginning of the root canal therapy. Clinical and radiographic exams were performed to evaluate the presence of PAP or healthy periradicular tissues (healed). Saliva samples were collected as a genomic DNA. The genotyping of ESR1 (rs2234693 and rs9340799), ESR2 (rs1256049 and rs4986938), VDR (rs739837 and rs2228570) and miRNA17 (rs4284505) were performed by real-time PCR. Chi-square test was used to the distribution of genotype and allele frequencies. Haplotype analysis was also performed. Eighty-nine patients were included in the “healed” group and 73 in the “PAP” group. No association was found between the allelic and genotypic polymorphisms studied and PAP (p>0.05). Haplotype analysis also did not demonstrated an association (p>0.05). In conclusion, the genetic polymorphisms in ESR1, ESR2, VDR and miRNA17 are not associated with PAP.

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.

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.

Resumo O objetivo deste estudo foi avaliar o reparo periapical e a síntese de mediadores inflamatórios após tratamento endodôntico em dentes de cães com lesão periapical, em sessão única, utilizando diferentes protocolos de irrigação. Lesões periapicais foram induzidas experimentalmente em dentes de cães e aleatoriamente divididas em 4 grupos: G1 - Irrigação por Pressão Apical Negativa (n = 20); G2 - Irrigação Ultrassônica Passiva (n = 20), G3 - Irrigação por Pressão Positiva (n = 20); G4 - Lesão periapical sem tratamento (n = 20). Após 180 dias, os animais foram eutanasiados, as peças removidas e submetidas ao processamento histotécnico para análise imunohistoquímica para osteopontina (OPN), fator de necrose tumoral-a (TNF-a) e interleucina 1-a (IL-1a). A análise radiográfica do reparo das lesões periapicais foi realizada por meio do Índice Periapical, obtido antes e 180 dias após o tratamento endodôntico. Os resultados obtidos foram submetidos à análise estatística por meio dos testes de sinais de Wilcoxon, Exato de Fisher ou Kruskal-Wallis seguido pelo pós-teste de Dunn (a = 5%). O exame radiográfico após o tratamento endodôntico, mostrou a persistência de áreas radiolúcidas periapicais e descontinuidade da lâmina dura em 35% dos espécimes do G1, 40% do G2 e 40% do G3, embora uma redução no PAI tenha sido observada (p<0,05). Pela análise imuno-histoquímica, o tratamento endodôntico resultou na menor síntese de TNF-a e de OPN na região periapical, comparativamente à lesão periapical sem tratamento (p<0,05). A produção de IL-1a não foi modulada pelo tratamento endodôntico (p>0,05). Reparo da lesão periapical foi observado em cerca de 60% dos casos após tratamento endodôntico realizado em sessão única e menor síntese de TNF-a e de OPN na região periapical, independente do protocolo de irrigação utilizado.

Abstract The aim of this study was to evaluate the radiographic periapical repair and the synthesis of inflammatory mediators after endodontic treatment in a single session, using different irrigation protocols, in teeth with apical periodontitis. Experimental apical periodontitis were induced in dog’s teeth randomly assigned into 4 groups: G1 - Irrigation by Negative Apical Pressure (n= 20); G2 - Passive Ultrasonic Irrigation (n= 20), G3 - Positive Pressure Irrigation (n= 20); G4 - apical periodontitis without treatment (n= 20). After 180 days, the animals were euthanized, the tissues removed and submitted to histotechnical processing for immunohistochemical analysis of osteopontin (OPN), tumor necrosis factor-a (TNF-a) and interleukin 1-a (IL-1a). Radiographic analysis was performed using the Periapical Index (PAI), obtained prior to and 180 days following endodontic treatment. Data were analyzed using Wilcoxon signed-rank test, Fisher’s Exact test or Kruskal-Wallis test and Dunn’s post-test (a = 5%). Radiographically, after endodontic treatment, apical periodontitis persisted in 35% of G1 specimens, 40% of G2 and 40% of G3 (p>0.05), although a PAI reduction was observed (p<0.05). By immunohistochemical evaluation, endodontic treatment resulted in lower synthesis of TNF-a and OPN in periapical region, compared to apical periodontitis without treatment (p<0.05). Production of IL-1 was not modulated by endodontic treatment (p>0.05). Periapical healing was observed in approximately 60% of the cases after endodontic treatment performed in a single session with lower synthesis of TNF-a and OPN in the periapical region, regardless of the irrigation protocol used.

Abstract Objectives: Infection, inflammation and bone resorption are closely related events in apical periodontitis development. Therefore, we sought to investigate the role of cyclooxygenase (COX) in osteoclastogenesis and bone metabolism signaling in periapical bone tissue after bacterial lipopolysaccharide (LPS) inoculation into root canals. Methodology: Seventy two C57BL/6 mice had the root canals of the first molars inoculated with a solution containing LPS from E. coli (1.0 mg/mL) and received selective (celecoxib) or non-selective (indomethacin) COX-2 inhibitor. After 7, 14, 21 and 28 days the animals were euthanized and the tissues removed for total RNA extraction. Evaluation of gene expression was performed by qRT-PCR. Statistical analysis was performed using analysis of variance (ANOVA) followed by post-tests (α=0.05). Results: LPS induced expression of mRNA for COX-2 (Ptgs2) and PGE2 receptors (Ptger1, Ptger3 and Ptger4), indicating that cyclooxygenase is involved in periapical response to LPS. A signaling that favours bone resorption was observed because Tnfsf11 (RANKL), Vegfa, Ctsk, Mmp9, Cd36, Icam, Vcam1, Nfkb1 and Sox9 were upregulated in response to LPS. Indomethacin and celecoxib differentially modulated expression of osteoclastogenic and other bone metabolism genes: celecoxib downregulated Igf1r, Ctsk, Mmp9, Cd36, Icam1, Nfkb1, Smad3, Sox9, Csf3, Vcam1 and Itga3 whereas indomethacin inhibited Tgfbr1, Igf1r, Ctsk, Mmp9, Sox9, Cd36 and Icam1. Conclusions: We demonstrated that gene expression for COX-2 and PGE2 receptors was upregulated after LPS inoculation into the root canals. Additionally, early administration of indomethacin and celecoxib (NSAIDs) inhibited osteoclastogenic signaling. The relevance of the cyclooxygenase pathway in apical periodontitis was shown by a wide modulation in the expression of genes involved in both bone catabolism and anabolism.