Cytotoxicity of substances leached from a root canal sealer based on mineral trioxide aggregate

Objetivo: Com base nas características biológicas e físico-químicas do agregado de trióxido mineral (MTA), este seria o material mais adequado para a obturação do canal radicular. No entanto, esse material apresenta baixo escoamento e, consequentemente, difícil manipulação. O MTA Fillapex (MTA-F) foi criado na tentativa de combinar as propriedades físico-químicas do cimento endodôntico com as propriedades biológicas do MTA. No entanto, os estudos sobre as características biológicas do MTA-F ainda são controversos. Dessa forma, este estudo teve como objetivo analisar in vitro a citotoxicidade do MTA-F. Materiais e Métodos: fi broblastos gengivais foram cultivados em Dulbecco’s modifi ed Eagle Medium (DMEM) e submetidos ao meio de cultura condicionado pelo MTA ou MTA-F. Esse meio condicionado continha substâncias liberadas pelos cimentos endodônticos. Células cultivadas em meio fresco serviram como controle positivo. A viabilidade celular foi avaliada por ensaio do MTT após 1, 3, 5 e 7 dias. Os dados obtidos foram comparados por análise de variância (ANOVA) seguida pelo teste de Tukey (p < 0,05). Resultados: As células submetidas ao meio condicionado pelo MTA apresentaram curva de crescimento celular semelhante à das células do grupo controle. Para o grupo MTA-F, não houve crescimento celular e foi observado um número de células viáveis significativamente menor do que o dos demais grupos durante todo o experimento. Conclusão: Substâncias liberadas a partir de MTA-F não permitiram o crescimento celular, mostrando que esse cimento endodôntico à base de MTA é altamente citotóxico. A característica de biocompatibilidade do MTA pode ser perdida com o MTA-F e comprometer o sucesso do tratamento endodôntico.Aim: Based on its biological and physicochemical characteristics, mineral trioxide aggregate (MTA) could be considered the most appropriate material for root canal obturation; nevertheless, handling of MTA is not easy. The MTA Fillapex (MTA-F) was created in an attempt to combine the physicochemical properties of a root canal sealer with the biological properties of MTA. However, the studies on the biological characteristics of MTA-F are still controversial. Thus, this study aimed to analyze the cytotoxicity of MTA-F. Materials and Methods: Cultured human gingival fi broblasts were grown in Dulbecco’s modifi ed Eagle Medium (DMEM) and submitted to a cell culture medium conditioned by MTA or MTA-F. The conditioned medium contained substances leached from the root canal sealers. Cells grown on a fresh medium served as a positive control. Cell viability was assessed by MTT assay at 1, 3, 5 and 7 days. Data was compared by ANOVA followed by Tukey’s test (p < 0.05). Results: Cells submitted to media conditioned by MTA presented a cell growth curve similar to that of the control cells. For the MTA-F group, cell growth was not observed and cell viability was signifi cantly lower than for the other groups during the entire experiment. Conclusion: Substances leached from MTA-F did not allow cell growth, indicating that this MTA-based root sealer is highly cytotoxic. The biocompatibility characteristic of MTA can be lost with MTA-F, and may compromise the endodontic treatment outcome

Second Mesiobuccal Root Canal of Maxillary First Molars in a Brazilian Population in High-Resolution Cone-Beam Computed Tomography

Introduction: The second canal of the mesiobuccal root (MB2) of the maxillary first molars (MFM) is difficult to detect in conventional radiographs and can be a major cause of failure in endodontic treatments. The aim of this study was to investigate the prevalence and anatomy of the MB2 by using high-resolution cone-beam computed tomography (CBCT). Methods and Materials: Three radiologists examined 414 high-resolution CBCTs. Of these, the CBCTs of 287 patients (mean age 49.43±16.76) who had at least one MFM were selected, making a total of 362 teeth. Prevalence and its relation with gender and age of the patients, side of the tooth, and Vertucci’s classification were analyzed. Data were statistically analyzed (P<0.05). Results: A total of 68.23% of the teeth exhibited the MB2. The presence of the MB2 was equivalent in both genders and significantly higher in younger patients. There was no correlation between the presence of the MB2 in relation to both the sides of the MFM and the FOV size. Smaller FOV recognized higher Vertucci’s grades. Conclusions: It was concluded that the prevalence of the MB2 canal in maxillary first molars in this Brazilian population examined with high-resolution CBTCs is 68.23%, being more prevalent in young patients. Gender and the side examined are no factors for determining the presence of MB2. Although the both FOVs of the high-resolution CBTCs (FOV 8 and 5) detect the MB2 canal, smaller FOV (FOV 5) is more accurate in the analysis of the internal anatomy of such root canals, according to the Vertucci´s classification.Keywords: Cone-beam Computed Tomography; High Resolution; Maxillary First Molar ; Mesiobuccal Root; High Resolutio

Dental Pulp Vascular Permeability Changes Induced by Dental Bleaching

Aiming to compare the effect of different light sources for dental bleaching on vascular permeability of dental pulps, forty-eight incisors were used. The bleaching agent (35% hydrogen peroxide) was activated by halogen light; LED (Light Emitting Diode) or LED, followed by laser phototherapy (LPT) (lambda = 780 nm; 3 J/cm(2)). After the bleaching procedures, the animals received an intra-arterial dye injection and one hour later were sacrificed. The teeth were diaphanized and photographed. The amount of blue stain content of each dental pulp was quantified using a computer imaging program. The data was statistically compared (p <= 0.05). The results showed a significant higher (p <= 0.01) dye content in the groups bleached with halogen light, compared with the control, LED and LED plus LPT groups. Thus, tooth bleaching activated by LED or LED plus LPT induces lesser resulted in increased vascular permeability than halogen light

Analysis of ECM proteins and MMPs expression in human dental pulp stem cells

Células-tronco adultas podem ser isoladas de vários tecidos, dentre eles a polpa dentária humana, tecido originado na papila dentária do dente em desenvolvimento. Estas linhagens multipotentes podem ser estudadas sob vários aspectos, como na elucidação da histogênese de tumores. O objetivo deste estudo foi inferir a histogênese do mixoma odontogênico, neoplasia odontogênica benigna, analisando a expressão de proteínas da matriz extracelular (MEC) e de metaloproteinases da matriz (MMPs) em células-tronco adultas de polpa dentária humana. Três linhagens diferentes de células-tronco originadas de polpas dentárias humanas IDPSCs (DL-1, DL-2 e DL4) foram utilizadas. As proteínas analisadas foram as mesmas expressas na neoplasia: vimentina, colágeno tipo I, fibronectina, tenascina, ácido hialurônico e MMPs (MMP-1, MMP-2 e MMP-9). Imunofluorescência e ensaios enzimáticos foram utilizados para analisar a presença de proteínas nas células cultivadas e no meio de cultura condicionado por estas células, respectivamente. Todas as linhagens celulares expressaram a vimentina e nenhuma expressou o ácido hialurônico. A linhagem celular DL-1 expressou todas as outras proteínas da matriz extracelular estudadas, enquanto que na linhagem DL-2 apenas não foi observada a expressão do colágeno tipo I. Fibronectina e tenascina não foram observados na linhagem DL-4. Todas as linhagens expressaram todas as MMPs, sendo que a produção de MMP-2 nas três linhagens foi significantemente maior que a de todas outras MMPs. Baseado nas condições deste estudo, é possível concluir que a expressão de proteínas da MEC e de MMPs em células-tronco de polpa dentária humana apresentaram perfil similar àquela apresentada no mixoma odontogênico, exceto pela ausência de marcação do ácido hialurônico em todas as linhagens. A ausência de secreção de ácido hialurônico pelas IDPSCs poderia indicar que o mixoma odontogênico deriva de uma célula mais diferenciada que as células-tronco.Adult stem cells can be isolated from different tissues including the human dental pulp, a structure originated from the dental papillae. These cell lineages are of importance in a series of studies, as the analysis of tumors histogenesis. The aim of this study was to infer the histogenesis of odontogenic myxoma, a benign odontogenic neoplasia by analyzing the ECM and MMPs molecules expressed in human dental pulp stem cells. Three different lineages of immature dental pulp stem cells (IDPSCs) (DL-1, DL-2 and DL-4) were used. The proteins searched were those expressed by the tumoral cells: vimentin, type I collagen, fibronectin, tenascin and hialuronic acid (HA) and the matrix metalloproteinases (MMP-1, MMP-2 and MMP-9). Immunofluorecence and enzymatic assays were used for analyzing the presence of the proteins in the cells and in the culture media conditioned by the cells, respectively. All the lineages expressed vimentin; however none expressed HA. DL-1 lineage expressed all the other ECM proteins, and the expression of type I collagen was not observed in the DL-2 lineage. Fibronectin and tenascin were not observed in the DL-4 lineage. All the lineages expressed all the MMPs. The release of MMP-2 from all cell lineages was significantly higher than those of all other MMPs. Based on the conditions of this study its possible to conclude that the overall expression of MEC proteins and MMPs in the lineages of human dental pulp stem cells were similar to those found in the odontogenic myxoma, except for the absence of hyaluronic acid. The absence of HA secretion by the IDPSCs could indicate that the odontogenic myxoma tumoural cells derive from a cell more differentiated than the stem cells

Dental pulp stem cells express proteins involved in the local invasiveness of odontogenic myxoma

Little is known about the histogenesis of the odontogenic myxoma (OM). Dental pulp stem cells could be candidate precursors of OM because both OM and the dental pulp share the same embryological origin: the dental papilla. For the purpose of comparing OM and stem cells, this study analyzed the expression of two proteins related to OM invasiveness (MMP-2 and hyaluronic acid) in human immature dental pulp stern cells (hIDPSCs). Three lineages of hIDPSCs from deciduous and permanent teeth were used in this study. Immunofluorescence revealed positive reactions for MMP-2 and hyaluronic acid (HA) in all hIDPSCs. MMP-2 appeared as dots throughout the cytoplasm, whereas HA appeared either as diffuse and irregular dots or as short fibrils throughout the cytoplasm and outside the cell bodies. The gene expression profile of each cell lineage was evaluated using RT-PCR analysis, and HA was expressed more intensively than MMP-2. HA expression was similar among the three hIDPSCs lineages, whereas MMP-2 expression was higher in DL-1 than in the other cell lines. The expression of proteins related to OM invasiveness in hIDPSCs could indicate that OM originates from dental pulp stem cells

Effect of laser phototherapy on the release of fibroblast growth factors by human gingival fibroblasts

The effects of laser phototherapy on the release of growth factors by human gingival fibroblasts were studied in vitro. Cells from a primary culture were irradiated twice (6 h interval), with continuous diode laser [gallium-aluminum-arsenium (GaAlAs), 780 nm, or indium-gallium-aluminum-phosphide (InGaAlP),_660 nm] in punctual and contact mode, 40 mW, spot size 0.042 cm(2), 3 J/cm(2) and 5 J/cm(2) (3 s and 5 s, respectively). Positive [10% fetal bovine serum (FBS)] and negative (1%FBS) controls were not irradiated. Production of keratinocyte growth factor (KGF) and basic fibroblast growth factor (bFGF) was quantified by enzyme-linked immunosorbent assay (ELISA). The data were statistically compared by analysis of variance (ANOVA) followed by Tukey`s test (P a parts per thousand currency signaEuro parts per thousand 0.05). The characterization of the cell line indicated a mesenchymal nature. KGF release was similar in all groups, while that of bFGF was significantly greater (1.49-times) in groups treated with infra-red laser. It was concluded that increased production of bFGF could be one of the mechanisms by which infra-red laser stimulates wound healing

Expression of Extracellular Matrix Proteins in Human Dental Pulp Stem Cells Depends on the Donor Tooth Conditions

Introduction: Stem cells are characterized by the ability to renew themselves through mitotic cell division and differentiating into a diverse range of specialized cell types. An important source of adult stem cells is the dental pulp. In dentistry, regenerative strategies are of importance because of hard dental tissue damage especially as result of caries lesions, trauma, or iatrogenic procedures. The regeneration of dental tissues relies on the ability of stem cells to produce extracellular (ECM) proteins encountered in the dental pulp tissue. Thus, the aim of this study was to analyze the expression and distribution of proteins encountered in dental pulp ECM (type I collagen, fibronectin, and tenascin) in stem cells. Methods: Human immature dental pulp stem cells (hIDPSCs) from deciduous (DL-1 and DL-4 cell lines) and permanent (DL-2) teeth were used. The distribution of ECM proteins was observed using the immunofluorescence technique. The gene expression profile was evaluated using reverse transcription polymerase chain reaction (RT-PCR) analysis. Results: Positive reactions for all ECM proteins were observed independently of the hIDPSCs analyzed. Type I collagen appeared less evident in DL-2 than in other hIDPSCs. Fibronectin and tenascin were less clear in DL-4. The RT-PCR reactions showed that type I collagen was lesser expressed in the DL-2 cells, whereas fibronectin and tenascin were similarly expressed in all hIDPSCs. Conclusions: The distribution and expression of ECM proteins differ among the hIDPSCs. These differences seemed to be related to the donor tooth conditions (deciduous or permanent, retained or erupted, and degree of root reabsorption). (J Endod 2010;36:826-831)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Desenmvolvimento Cientifico e Tecnologico (CNPq)Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (Capes