Temporo-spatial distribution of stem cell markers CD146 and p75NTR during odontogenesis in mice

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

Phototherapy With LED as an Effective Treatment for Chemotherapy-Induced Oral Mucositis in Hamsters

Introduction: Oral mucositis (OM) has been considered one of the most feared collateral effects of oncological treatments. Some therapies have been used, such as light-emitting diode (LED), with promising results, but with no sufficient evidence in the literature.Objective: Our study aimed to evaluate, by clinical and histological analysis, the effect of LED on the treatment of chemotherapy-induced OM (CIOM) in an animal model.Methods: Twenty male hamsters were equally distributed to two groups: control (C), which received anesthesia and CIOM induction; and LED (L), which received anesthesia, CIOM induction, and LED treatment (635 nm, 120 mW, 0.48 J). The clinical analysis was performed through two specific scales for OM analysis on days 5, 7 and 10 of the experiment. In addition, the injured area of all hamsters check pouch mucosa was removed and processed for histological analysis on the last experimental day.Results: After statistical analysis, group L showed less severity of OM when compared with the C group (P < 0.05); beyond that, both healed completely on day 10.Conclusion: Our results suggested that the phototherapy with LED had a positive effect on accelerating repair, reducing the severity of CIOM

Aderência de Actinobacillus actinomycetemcomitans às células epiteliais bucais: estabilidade e aspectos ultra-estruturais

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.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

Immunohistological study of the effect of vascular Endothelial Growth Factor on the angiogenesis of mature root canals in rat molars

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

Bond strength and morphology of enamel using self-etching adhesive systems with different acidities

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 (

Effect of fluoride application during radiotherapy on enamel demineralization

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

Pulp analysis of teeth submitted to different types of forces: a histological study in rats

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. &nbsp

In vitro evaluation of hydroxyapatite, chitosan, and carbon nanotube composite biomaterial to support bone healing

O compósito à base de hidroxiapatita, quitosana e nanotubo de carbono foi desenvolvido com o intuito de auxiliar na consolidação óssea. Estudos anteriores sugerem que a combinação de substitutos ósseos e células-tronco mesenquimais (CTM) podem auxiliar a potencializar e promover a regeneração óssea. No presente estudo, o biomaterial foi desenvolvido e a viabilidade e a citotoxicidade de células Vero (ATCC CCL-81) e CTM obtidas de medula óssea provenientes de ovinos utilizando ensaios metil-tiazol-tetrazólio, MTT e iodeto de propídeo (PI) foram avaliadas em diferentes concentrações de extrato desse compósito. O compósito demonstrou ausência de citotoxicidade com comportamento semelhante ao grupo controle. Amostras com 50% e 10% de concentração de extrato do compósito mostraram resultados maiores comparados ao grupo controle (ensaio MTT). Esses resultados também sugerem que a presença do biomaterial pode ser utilizada em associação a CTM. Assim, esse estudo conclui que o compósito apresentado de hidroxiapatita, quitosana e nanotubo de cabono não foi considerado citotóxico e pode ser utilizado em teste in vivo.Hydroxyapatite, chitosan, and carbon nanotube composite biomaterial were developed to improve bone healing. Previous studies suggested that a combination of biomaterials and mesenchymal stem cells (MSCs) can potentially help promote bone regeneration. In the present study, we first developed hydroxyapatite, chitosan, and carbon nanotube composite biomaterial. Then, the effect of different concentrations of the extract on the viability of Vero cells (ATCC CCL-81) and MSCs obtained from sheep bone marrow using methylthiazol tetrazolium (MTT) and propidium iodide (PI) assays were evaluated. The biomaterial group demonstrated an absence of cytotoxicity, similar to the control group. Samples with 50% and 10% biomaterial extract concentrations showed higher cell viability compared to samples from the control group (MTT assay). These results suggest that the presence of this composite biomaterial can be used with MSCs. This study also concluded that hydroxyapatite, chitosan, and carbon nanotube composite biomaterial were not cytotoxic. Therefore, these could be used for performing in vivo tests

Effects of a non-rinse conditioner on the enamel of primary teeth

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