Abstract Mesenchymal and epithelial stem cells were identified in dental tissues; however, knowledge about the odontogenic stem cells is limited, and there are some questions regarding their temporo-spatial dynamics in tooth development. Objective Our study aimed to analyze the expression of the stem cell markers CD146 and p75NTR during the different stages of odontogenesis. Methodology The groups consisted of 13.5, 15.5, 17.5 days old embryos, and 14 days postnatal BALB/c mice. The expression of CD146 and p75NTR was evaluated by immunohistochemistry. Results Our results showed that positive cells for both markers were present in all stages of tooth development, and the number of positive cells increased with the progression of this process. Cells of epithelial and ectomesenchymal origin were positive for CD146, and the expression of p75NTR was mainly detected in the dental papilla and dental follicle. In the postnatal group, dental pulp cells were positive for CD146, and the reduced enamel epithelium and the oral mucosa epithelium showed immunostaining for p75NTR. Conclusions These results suggest that the staining pattern of CD146 and p75NTR underwent temporal and spatial changes during odontogenesis and both markers were expressed by epithelial and mesenchymal cell types, which is relevant due to the significance of the epithelial-ectomesenchymal interactions in tooth development.

Resumo O presente estudo tem como objetivo analisar o efeito da fototerapia LED na presença da hialinização e reabsorção radicular durante o movimento dentário ortodôntico (MDO) em ratos, e a mensuração da quantidade de movimento dentário. Oitenta ratos foram alocados em dois grupos: LED e Controle (CON), os ratos foram irradiados com um LED infravermelho (850nm, 30mW) por 5 minutos durante os cinco primeiros dias da MDO; e o grupo controle não foi irradiado. Ambos os grupos foram subdivididos em 4 subgrupos (n=10) de acordo com a data da eutanásia (4, 7, 14 e 21 dias). Cinco dos dez ratos LED21 e cinco dos dez ratos CON21foram submetidos a microtomografia computadorizada (μCT); As μCT foram realizadas nos dias 0, 7, 14 e 21. Para o estudo histológico, as maxilas foram processadas para microscopia de luz, usando hematoxilina-eosina (HE) e Fosfatase ácido Tartrate-Resistente (TRAP) para histoquímica. A quantidade de movimento dentário não diferiu entre o LED e o CON. A hialinização foi observada nas áreas de pressão em ambos os grupos e não mostrou diferença estatisticamente significante. Reabsorção radicular também foi observada em ambos os grupos depois de 7 dias e não houve diferença entre os grupos. A fototerapia LED não aumentou a quantidade de MDO. Características similares de hialinização e reabsorção radicular foram observadas em ambos os grupos

Abstract The present study aimed to analyze the effect of LED phototherapy on the presence of hyalinization and root resorption during orthodontic tooth movement (OTM) in rats and to measure the amount of tooth movement. Eighty rats were allocated into two groups: LED and control (CON), where the LED rats were irradiated with infrared LED (850 nm, 30 mW) for 5 min during the first five days of OTM and where controls were not irradiated. Both groups were subdivided into four subgroups (n=10) according to the date of euthanasia (4, 7, 14 and 21 days). Five out of ten LED21 and five of ten CON21 rats were submitted to micro-computed tomography (μCT); μCT scans were taken on days 0, 7, 14 and 21. For histological study, maxillae were processed to light microscopy using Hematoxylin-Eosin (HE) and Tartrate-Resistant Acid Phosphatase (TRAP) histochemistry. The amount of tooth movement did not differ between LED and CON. Hyalinization was observed at the pressure areas in both groups, and it did not show a statistically significant difference between the groups. Root resorption was also observed in both groups after 7 days and it did not represent any differences between the two groups. LED phototherapy was not able to increase the amount of OTM. Similar characteristics of hyalinization and root resorption were observed in both groups.

Abstract Radiation-related caries are one the most undesired reactions manifested during or after head and neck radiotherapy. Fluoride application is an important strategy to reduce demineralization and enhance remineralizaton. Objective: To evaluate the effect of the topical application of fluoride during irradiation on dental enamel demineralization. Material and Methods: Thirty molars were randomly divided into three groups: Non-irradiated (NI), Irradiated (I), Irradiated with fluoride (IF). Each group was subdivided according to the presence or absence of pH-cycling (n=5). In the irradiated groups, the teeth received 70 Gy. The enamel's chemical composition was measured using Fourier Transform Infrared Spectrometry (organic matrix/mineral ratio - M/M and relative carbonate content - RCC). Vickers microhardness (VHN) and elastic modulus (E) were evaluated at three depths (surface, middle and deep enamel). Scanning electron microscopy (SEM) was used to assess the enamel's morphology. Results: The FTIR analysis (M/M and RCC) showed significant differences for irradiation, pH-cycling and the interaction between factors (p<0.001). Without pH-cycling, IF had the lowest organic matrix/mineral ratio and relative carbonate content. With pH-cycling, the organic matrix/mineral ratio increased and the relative carbonate content decreased, except for IF. VHN was influenced only by pH-cycling (p<0.001), which generated higher VHN values. ANOVA detected significant differences in E for irradiation (p<0.001), pH-cycling (p<0.001) and for the interaction between irradiation and pH-cycling (p<0.001). Increased E was found for group I without pH-cycling. With pH-cycling, groups I and IF were similar, and showed higher values than NI. The SEM images showed no morphological changes without pH-cycling. With pH-cycling, fluoride helped to maintain the outer enamel's morphology. Conclusions: Fluoride reduced mineral loss and maintained the outer morphology of irradiated and cycled enamel. However, it was not as effective in preserving the mechanical properties of enamel. Radiotherapy altered the enamel's elastic modulus and its chemical composition.

Abstract Tissue bioengineering has been applied to Endodontics to seek a more biological treatment. The presence of blood vessels is crucial for cell nutrition during tissue formation. Objective This study analysed the application of vascular endothelial growth factor (VEGF) in the angiogenesis of mature root canals. Material and methods Upper first molars of twelve 13-week old Wistar male rats were used. The root pulp of the mesiobuccal canal was removed and the root canal instrumented with K-files up to size #25. Periapical bleeding was induced into the root canal by introducing a #15 K-file beyond the apex. The teeth on the right side of the arch were filled up with blood clot (G1), whereas those on the left side were filled up with blood clot plus 50 ng/ml of VEGF (G2). Teeth were sealed with light-curing glass-ionomer cement and the animals were sacrificed after 60 days. The maxilla was dissected and fixed before obtaining serial sections for histological processing with haematoxylin-eosin (HE) and immunohistochemical factor-VIII. Immunohistochemical labelling was evaluated using scores for statistical analysis. Results Immunohistological analysis demonstrated the presence of angiogenesis in both groups, but with higher angiogenic maturation in G2 during the experimental period (p<0.05). HE staining showed connective tissue with absence of odontoblasts in all specimens. Conclusions It can be concluded that it is possible to obtain angiogenesis in mature root canals with or without the use of VEGF, although the latter tends to accelerate blood vessel formation.

Abstract Objective: The purpose of this study was to histologically evaluate pulp and dentin under induced tooth movement (ITM) with different types of forces. Material and Methods: The maxillary right first molars of rats were submitted to movement with continuous (CF), continuous interrupted (CIF) and intermittent (IF) forces during 5, 7 and 9 days with nickel-titanium (NiTi) closed-coil springs exerting 50cN force magnitude. The groups were histologically evaluated as for cellularity pattern, presence of dystrophic, hemodynamic alterations in the pulp as well dentin alterations. The main observed alterations were related to hemodynamic pulp characteristics, such as presence of thrombosis, vascular congestion and hemorrhages. The hemodynamic alterations were statistically evaluated by Shapiro-Wilk normality test and analysis of variance by the Kruskall-Wallis test. Results: There was no significant differences observed between groups in the different types of applied forces and duration of ITM (vascular congestion, p=1.000; hemorrhage, p=0.305; thrombosis, p=1.000). Conclusions: Pulp tissue alterations resulting from ITM were limited to hemodynamic events, without progressing to irreversible degeneration, regardless of the type of force applied.

Resumo Este estudo foi conduzido para avaliar os efeitos do tratamento com ranelato de estrôncio (RE) na reparação de defeitos ósseos e componentes moleculares de ossos nos fêmures. Ratas adultas (n = 27) foram submetidas a ovariectomia (OVX) ou cirurgia Sham. Trinta dias após a cirurgia, um defeito foi feito no fêmur e os animais foram então divididos em três grupos: OVX, SHAM e OVX+RE. A eutanásia foi realizada quatro semanas após a cirurgia de preparo do defeito ósseo. A reparação do defeito ósseo foi avaliada por microtomografia computorizada (μCT) e a composição química do osso cortical foi analisada por espectroscopia de infravermelho de transformada de Fourier (FTIR) e espectroscopia por energia dispersiva de raios X (EDS). A espessura do osso trabecular (Tb.Th) recém formado no grupo OVX+SR foi significativamente maior que a do grupo OVX. A maturidade do colágeno no grupo OVX+SR foi menor do que nos outros dois grupos. Neste grupo, observou-se também um aumento significativo na quantidade de estrôncio (Sr) e uma diminuição na quantidade de cálcio (Ca) no tecido ósseo. O tratamento sistêmico com RE melhorou a microarquitetura do osso recém formado dentro do defeito, mas diminuiu a reticulação do colágeno maduro no osso cortical.

Abstract This study was conducted to evaluate the effects of treatment with strontium ranelate (SR) on the repair of bone defects and molecular components of bones in femurs. Adult female rats (n=27) were subjected to ovariectomy (OVX) or Sham surgery. Thirty days after surgery, a defect was made in the femur and the animals were then divided into three groups: OVX, SHAM and OVX+SR. Euthanasia was performed four weeks after the bone defect surgery. Repair in bone defect was assessed by computed microtomography (μCT) and chemical composition of cortical bone was analyzed by Fourier transform infrared (FTIR) spectroscopy and energy dispersive X-ray spectroscopy (EDS). The trabecular thickness (Tb.Th) of the newly formed bone in the OVX+SR group was significantly higher than that for the OVX group. The collagen maturity in the OVX+SR group was smaller than in the other two groups. In this group, a significant increase in the amount of strontium (Sr) and a decrease in the amount of calcium (Ca) embedded to bone tissue were also observed. Systemic treatment with SR improved microarchitecture of the newly formed bone inside the defect, but decreased cross-linking of mature collagen in cortical bone.

OBJECTIVES: To assess the bond strength and the morphology of enamel after application of self-etching adhesive systems with different acidities. The tested hypothesis was that the performance of the self-etching adhesive systems does not vary for the studied parameters. MATERIAL AND METHODS: Composite resin (Filtek Z250) buildups were bonded to untreated (prophylaxis) and treated (bur-cut or SiC-paper) enamel surfaces of third molars after application of four self-etching and two etch-and-rinse adhesive systems (n=6/condition): Clearfil SE Bond (CSE); OptiBond Solo Plus Self-Etch (OP); AdheSe (AD); Tyrian Self Priming Etching (TY), Adper Scotchbond Multi-Purpose Plus (SBMP) and Adper Single Bond (SB). After storage in water (24 h/37°C), the bonded specimens were sectioned into sticks with 0.8 mm² cross-sectional area and the microtensile bond strength was tested at a crosshead speed of 0.5 mm/min. The mean bond strength values (MPa) were subjected to two-way ANOVA and Tukey's test (α=0.05). The etching patterns of the adhesive systems were also observed with a scanning electron microscope. RESULTS: The main factor adhesive system was statistically significant (p<0.05). The mean bond strength values (MPa) and standard deviations were: CSE (20.5±3.5), OP (11.3±2.3), AD (11.2±2.8), TY (11.1±3.0), SBMP (21.9±4.0) and SB (24.9±3.0). Different etching patterns were observed for the self-etching primers depending on the enamel treatment and the pH of the adhesive system. CONCLUSION: Although there is a tendency towards using adhesive systems with simplified application procedures, this may compromise the bonding performance of some systems to enamel, even when the prismless enamel is removed.

A limpeza das paredes cavitárias é um passo importante na clínica odontológica e visa otimizar os procedimentos adesivos e a longevidade das restaurações. O presente estudo comparou o efeito de três agentes de limpeza cavitária - jato abrasivo de bicarbonato de sódio/ar/água (Profi II, Dabi Atlante, São Paulo, Brasil); pasta de pedra-pomes e água, somada a um detergente biológico (Tergestesim, Probem, São Paulo, Brasil); e spray de ar/água - na resistência adesiva entre dentina e dois tipos de adesivos dentais: Clearfil SE Bond ("self-etching") (Kuraray, Kioto, Japão) e Scotchbond Multi-Purpose Plus ("all-etching") (3M-ESPE, São Paulo, Brasil). Seis grupos de espécimes (n = 10) obtidos a partir de elementos dentais humanos extraídos por indicação foram preparados e cada um recebeu um dos tratamentos de superfície. Após aplicação dos adesivos, uma porção tronco-cônica de resina composta (AP-X, Kuraray, Kioto, Japão/Z-100; 3M-ESPE, São Paulo, Brasil) foi construída sobre os espécimes, com o auxílio de uma matriz bipartida de teflon e uma mesa metálica adaptadora para adaptação na máquina de ensaio de tração. Após armazenamento em água destilada a 37°C por 7 dias, os mesmos foram submetidos às provas de tração. Dois pares de cada grupo foram escolhidos aleatoriamente e processados para observação ao microscópio eletrônico de varredura (MEV). A análise estatística dos valores obtidos demonstrou que não houve diferenças significantes entre as técnicas de limpeza empregadas e nem entre a interação destas com os adesivos dentais, e as observações ao MEV revelaram uma predominância de fraturas mistas ocorridas na interface dentina/resina. Baseados nas análises estatísticas e nas observações ao MEV, concluiu-se que as técnicas de limpeza empregadas não interferem na resistência adesiva entre a dentina e os sistemas adesivos estudados, nas condições experimentais adotadas.

The cleaning of cavity walls aims to improve adhesive restorative procedures and longevity of restorations. This study has compared the effect of three cleaning agents - sodium bicarbonate jet (Profi II, Dabi Atlante, São Paulo, Brazil); pumice paste plus a biologic detergent (Tergestesim, Probem, São Paulo, Brazil); air water spray - on the bond strength between dentin and two different adhesive systems: Clearfil SE Bond (Kuraray, Kioto, Japan) and Scotchbond Multi-Purpose Plus (3M-ESPE, São Paulo, Brazil). Six groups (n:10) of dental fragments obtained from young adult extracted teeth were prepared, and each one received one of the listed surface cleaning techniques. After the adhesive application, a cone-shaped test body was built with AP-X (Kuraray, Kioto, Japan) or Z100 (3M-ESPE, São Paulo, Brazil) composite resins, using a Teflon matrix. The specimens were tested for tensile bond strength after one-week storage in distilled water at 37°C. Two pairs of fractured specimens of each group were randomly chosen and processed for scanning electron microscopy (SEM) analysis. ANOVA test of the bond strength values showed no statistical differences among the cleaning agents and neither between their interactions with the bonding systems. Upon SEM analysis, most surfaces showed mixed fractures of adhesive and cohesive failures in bonding resin to dentin. Based on statistical and SEM analysis, it was concluded that the cleaning agents studied did not interfere with the bond strength of the adhesive systems used to dentin.

O objetivo deste estudo in vitro foi de avaliar ao microscópio eletrônico de varredura os aspectos morfológicos do esmalte de dentes decíduos após condicionamento com ácido fosfórico a 36% ou com um agente condicionador não lavável. Foram selecionados 10 dentes decíduos anteriores esfoliados naturalmente. As amostras sofreram limpeza coronária com pasta de pedra-pomes e água, em baixa-velocidade. O condicionamento foi realizado no esmalte da face vestibular. Os espécimes foram divididos em dois grupos: G1 (n=10): condicionamento com ácido fosfórico a 36% na forma de gel - Conditioner 36 (Dentsply) durante 20 segundos, seguidos de lavagem com água durante 15 segundos; G2 (n=10): condicionamento com NRC - Non Rinse Conditioner (Dentsply) durante 20 segundos, seguidos de secagem com ar durante 15 segundos. As amostras foram desidratadas, montadas em bases metálicas e cobertas com ouro para análise ao microscópio eletrônico de varredura (Jeol JSM 6.100). A análise da eletromicrografias revelou que ambos os agentes condicionadores foram efetivos para condicionar o esmalte de dentes decíduos, causando microporosidades mas com melhor resultado quando utilizou-se o ácido fosfórico a 36% na forma de gel.

The aim of this in vitro study was to evaluate by scanning electron microscopy the morphological aspects of the enamel of primary teeth after etching with 36% phosphoric acid or a non-rinse conditioner. Ten naturally exfoliated anterior primary teeth were selected. The samples were subjected to prophylaxis with pumice paste and water using a low-speed hand piece. Etching was done on the buccal surface. Specimens were divided into 2 groups: G1 (n=10): etching with 36% phosphoric acid gel - Conditioner 36 (Dentsply) for 20 s, followed by water rinse for 15 s; G2 (n=10): etching with NRC - Non Rinse Conditioner (Dentsply) for 20 s, followed by air drying for 15 s. The samples were dehydrated, mounted on metal stubs, coated with gold and observed with Jeol JSM-6100 scanning electron microscope. Electron-micrographic analysis showed that both etching agents were effective for etching the enamel of primary teeth causing the formation of microporosities on the enamel surface, although the etching pattern was more effective with the use of 36% phosphoric acid gel.

Actinobacillus actinomycetemcomitans é considerado um importante patógeno na doença periodontal, particularmente na periodontite juvenil localizada. O mecanismo de adesão bacteriana às células epiteliais bucais (CEB), aos dentes e a outras bactérias, constitui-se o passo inicial na colonização e patogênese nos quadros de gengivite e periodontite. Neste estudo, avaliou-se a aderência às CEB, a sua variabilidade e os aspectos ultra-estruturais de 21 isolados e de uma cepa de referência de A. actinomycetemcomitans, quando submetidos a repiques sucessivos. Todos os isolados testados aderiram às CEB e os repiques sucessivos determinaram variações nas taxas de aderência de cada isolado. Os isolados que apresentaram altos índices de aderência também produziram quantidades elevadas de componentes extracelulares, tais como fímbrias, vesículas e/ou material amorfo extracelular.

Actinobacillus actinomycetemcomitans is considered an important pathogen in periodontal disease, especially in localized juvenile periodontitis. The bacterial mechanism of adherence to oral epithelial cells (OEC), teeth or other bacteria is the initial step in colonization and in the pathogenesis of oral infectious processes, such as gingivitis and periodontitis. The goal of this study was to evaluate the adherence and its variability, as well as ultrastructural aspects of A. actinomycetemcomitans isolates on OEC, when they were submitted to repeated subcultures. Twenty-one fresh isolates from 21 patients with periodontal disease and one reference strain were tested. All tested isolates adhered to OEC and repeated subcultures produced variations in the adhesion rates of each isolate. Furthermore, a direct correlation between high levels of adherence and the presence of large amounts of extracellular components, such as fimbriae, vesicles and extracellular amorfous material, was observed.