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.

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.

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 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.

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.

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.

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: 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 Objective: Avaliar a eficácia do XP-endo Finisher (XP) na remoção da smear layer em canais radiculares, comparando diferentes protocolos de irrigação. Métodos: Setenta e dois dentes humanos unirradiculares foram similarmente instrumentados usando limas R25 (VDW, Munich, Germany) aplicadas no modo reciprocrantes em um motor endodôntico VDW GOLD (VDW, Munich, Germany). O comprimento de trabalho foi determinado a 1 mm aquém do forame apical. Os canais foram irrigados com 5 mL de hipoclorito de sódio 2,5% durante a instrumentação. Os dentes foram divididos aleatoriamente em seis grupos (n=12). Um grupo controle, que não foi submetido ao protocolo final de irrigação, e cinco grupos experimentais com diferentes irrigantes e técnicas de agitação: EDTA/Lima, EDTA/XP, EDTA/Irrigação Ultrassônica Passiva (IUP), Água destilada (AD)/XP, e AD/IUP). Os escores de qualidade de remoção da camada de smear layer foram avaliados nos terços apical, médio e cervical do canal radicular com base em imagens obtidas por microscopia eletrônica de varredura. Os dados foram analisados pelo teste de Kruskal-Wallis, seguido de comparações dois a dois pelo teste de Dunn (α = 5%). Resultados: Os grupos EDTA/lima, EDTA/PUI e EDTA/XP demonstraram escores significativamente menores que os outros grupos (P <0,05). Não foi observada diferença entre os grupos que utilizaram água destilada e o grupo controle em todos os terços avaliados (P> 0.05). Conclusão: A lima XP-endo Finisher não aumentou a eficiência do EDTA na remoção da smear layer em canais radiculares.

ABSTRACT Objective: The aim of this study was to evaluate the effectiveness of XP-endo Finisher (XP) on removal of the smear layer in root canals by comparing different irrigation protocols. Methods: Seventy-two human single-rooted teeth were similarly instrumented using R25 Reciproc files (VDW, Munich, Germany) applied in reciprocating mode with a VDW GOLD endo motor (VDW, Munich, Germany). The working length was determined at 1 mm short of the apical foramen. The canals were irrigated with 5 mL of 2.5% sodium hypochlorite during instrumentation. The teeth were divided at random into six groups (n=12). A control group, which was not submitted to the final irrigation protocol, and five experimental groups with different irrigants and agitation techniques: EDTA/File, EDTA/XP, EDTA/Passive Ultrasonic Irrigation (PUI), Distilled Water (DW)/XP, and DW/PUI). Smear layer removal quality scores were assessed in the apical, middle, and cervical thirds of the root canal based on images obtained by scanning electron microscopy. Data were analyzed using the Kruskal-Wallis test, followed by two-by-two comparisons with the Dunn test (α=5%). Results: EDTA/File, EDTA/PUI, and EDTA/XP groups demonstrated significantly lower scores than the other groups (P<0.05) in all thirds evaluated. No significant difference was observed between the groups in which distilled water was used and the control group in all thirds evaluated (P> 0.05). Conclusion: The XP-endo Finisher file did not increase the efficiency of EDTA in removal of the smear layer in root canals.

Aim To evaluate the psychological impact of COVID-19 on undergraduate and graduate students of the Dental School of Ribeirão Preto, University of São Paulo. Methods Three questionnaires were used: sociodemographic, WHO Abbreviated Quality of Life Inventory, and General Anxiety Disorder-7. Data were analyzed using Graph Pad Prism 7a (α = 5%). Mann-Whitney or Kruskal-Wallis and Dunn post-tests were used for statistical comparisons. The Spearman test was used as the correlation test. Results 257 students responded to the online form that assessed their quality of life and anxiety level. On a scale from 1 to 100, with 100 being the best quality of life, the average obtained was 64.71 (± 13.36). In addition, 87.6% of the students rated their quality of life as good or very good. 74.7% reported good or very good health. The anxiety analysis resulted in an average of 10.04 (± 4.5), indicating moderate anxiety levels. There was an inversely proportional correlation between age and degree of anxiety (p = 0.008, r = -0.1628) and self-perceived learning and student commitment (r = 0.69). Conclusion Despite the good quality of life and the students’ good self-perception of health, they showed a moderate degree of anxiety during the social distancing caused by COVID-19, also demonstrating a decrease in interest and commitment during distance education.

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 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 A homeostasia entre cálcio e fósforo salivares é importante para a manutenção da saúde bucal. O objetivo deste estudo foi avaliar se polimorfismos genéticos no receptor de estrógeno alfa (ESR1), receptor de estrógeno beta (ESR2) e no microRNA17 (microRNA17) estão associados com os níveis salivares de cálcio e fósforo. Saliva de 276 crianças com 12 anos de idade foi coletada com estímulo mastigatório e os níveis de cálcio e fósforo foram determinados por Espectrofotometria de Massa. O DNA genômico foi extraído da saliva remanescente e os polimorfismos genéticos em ESR1 (rs12154178, rs1884051, rs9340799 e rs2234693), em ESR2 (rs4986938 e rs1256049) e no miRNA17 (rs4284505) foram genotipados pela reação em cadeia da polimerase em tempo real utilizando sondas TaqMan. As diferenças estatísticas nas distribuições alélicas e genotípicas entre “baixo” e “alto” níveis de cálcio e fósforo foram determinadas usando os testes qui-quadrado e teste exato de Fisher. As análises foram ajustadas por sexo (alfa de 5%). O polimorfismo rs9340799 em ESR1 foi o genótipo menos comum associado com altos níveis de cálcio (p=0,03). O alelo menos comum em ESR1 rs1884051 foi associado com baixos níveis de fósforo (p=0,005) e houve um excesso de heterozigotos para miRNA17 rs4284505 entre os indivíduos com baixos níveis de cálcio salivar (p=0,002), ambos ajustados pelo sexo. Este estudo fornece evidências de que polimorfismos genéticos em ESR1 e miRNA17 estão envolvidos na determinação dos níveis de cálcio e fósforo salivares.

Abstract Homeostasis between salivary calcium and phosphorus is important for maintaining oral health. The aim of this study was to evaluate if polymorphisms in ESR1 (Estrogen Receptor Alpha), ESR2 (Estrogen Receptor Beta) and miRNA17 (microRNA17) are associated with calcium and phosphorus levels in saliva. Saliva from 276 12-year-old children were collected by masticatory stimulation and calcium and phosphorus levels were determined by Mass Spectrometry. Genomic DNA was extracted from remaining saliva and genetic polymorphisms in ESR1 (rs12154178, rs1884051, rs9340799 and rs2234693), in ESR2 (rs4986938 and rs1256049) and in miRNA17 (rs4284505) were genotyped using TaqMan chemistry and a real-time PCR equipment. Statistical differences in genotype and allele distributions between ‘low’ and ‘high’ calcium and phosphorus levels were determined using chi-square or Fisher´s exact tests. The analysis was also adjusted by sex (alpha of 5%). ESR1 rs9340799 had the less common genotype associated with higher calcium levels (p=0.03). The less common allele of ESR1 rs1884051 was associated with lower phosphorus levels (p=0.005) and there was an excess of heterozygotes for miRNA17 rs4284505 among individuals with lower calcium levels (p=0.002), both adjusted by sex. This study provides evidence that genetic polymorphisms in ESR1 and miRNA17 are involved in determining salivary calcium and phosphorus levels.

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.