Methylmercury-induced cytotoxicity and oxidative biochemistry impairment in dental pulp stem cells: the first toxicological findings.

BACKGROUND: Methylmercury (MeHg) is a potent toxicant able to harm human health, and its main route of contamination is associated with the consumption of contaminated fish and other seafood. Moreover, dental amalgams are also associated with mercury release on human saliva and may contribute to the accumulation of systemic mercury. In this way, the oral cavity seems to be the primary location of exposure during MeHg contaminated food ingestion and dental procedures but there is a lack of literature about its effects on dental tissues and the impact of this toxicity on human health. In this way, this study aimed to analyze the effects of different doses of MeHg on human dental pulp stem cells after short-term exposure. METHODS: Dental pulp stem cells from human exfoliated deciduous teeth (SHED) were treated with 0.1, 2.5 and 5 µM of MeHg during 24 h. The MeHg effects were assessed by evaluating cell viability with Trypan blue exclusion assay. The metabolic viability was indirectly assessed by MTT reduction assay. In order to evaluate an indicative of antioxidant defense impairment, cells exposed to 0.1 and 5 µM MeHg were tested by measuring glutathione (GSH) level. RESULTS: It was observed that cell viability decreased significantly after exposure to 2.5 and 5 µM of MeHg, but the metabolic viability only decreased significantly at 5 µM MeHg exposure, accompanied by a significant decrease in GSH levels. These results suggest that an acute exposure of MeHg in concentrations higher than 2.5 µM has cytotoxic effects and reduction of antioxidant capacity on dental pulp stem cells.

Association between Tooth Loss and Stroke: A Systematic Review.

This systematic review aims to evaluate the association between tooth loss and stroke. The Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) guidelines were followed. The PECO strategy was used to limit the eligibility criteria. The following databases were used on searches: PubMed, Scopus, Web of Science, The Cochrane Library, LILACS and OpenGrey. We included observational studies performed in adults (Population), in which patients with tooth loss (Exposition) and patients without tooth loss (Comparison) were observed to investigate the association between tooth loss and stroke (Outcome). After searches, the results were submitted to a selection process, followed by data extraction, quality assessment and risk of bias evaluation. The certainty of the evidence was evaluated through GRADE approach. A total of 925 potential studies were retrieved by the searches and 9 were included in this review. Seven of the included articles described an association between tooth loss and stroke. Low risk of bias and a low certainty of the evidence were identified to all studies. The certainty of the evidence may be associated with the observational nature of the included studies. Even though an association between tooth loss and stroke was suggested, the low strength of the current evidence indicated the need for further investigations with a better methodological design to conclude this question.

Evaluation of Resin-Based Material Containing Copaiba Oleoresin (Copaifera Reticulata Ducke): Biological Effects on the Human Dental Pulp Stem Cells.

The purpose of this study was to analyze in vitro the biological effects on human dental pulp stem cells triggered in response to substances leached or dissolved from two experimental cements for dental pulp capping. The experimental materials, based on extracts from Copaifera reticulata Ducke (COP), were compared to calcium hydroxide [Ca(OH)2] and mineral trioxide aggregate (MTA), materials commonly used for direct dental pulp capping in restorative dentistry. For this, human dental pulp stem cells were exposed to COP associated or not with Ca(OH)2 or MTA. Cell cytocompatibility, migration, and differentiation (mineralized nodule formation (Alizarin red assay) and gene expression (RT-qPCR) of OCN, DSPP, and HSP-27 (genes regulated in biomineralization events)) were evaluated. The results showed that the association of COP reduced the cytotoxicity of Ca(OH)2. Upregulations of the OCN, DSPP, and HSP-27 genes were observed in response to the association of COP to MTA, and the DSPP and HSP-27 genes were upregulated in the Ca(OH)2 + COP group. In up to 24 h, cell migration was significantly enhanced in the MTA + COP and Ca(OH)2 + COP groups. In conclusion, the combination of COP with the currently used materials for dental pulp capping [Ca(OH)2 and MTA] improved the cell activities related to pulp repair (i.e., cytocompatibility, differentiation, mineralization, and migration) including a protective effect against the cytotoxicity of Ca(OH)2.

The effect of three different pulp capping cements on mineralization of dental pulp stem cells.

This study evaluated the osteogenic differentiation of human dental pulp stem cells in response to substances released by the pulp capping agents, Biodentine (BD), mineral trioxide aggregate (MTA) and two-paste calcium hydroxide cement (CHC), along with their physicochemical characteristics. The dimensional stability test showed that of the materials studied, only BD met the standards recommended by the International Organization for Standardization (ISO) for pulp capping materials and thus can be used safely. In the chemical tests, BD was the most stable material. In the Alizarin red S test, BD formed the higher amount of mineralized nodules in the mineralizing medium and also formed mineralized nodules in a non-mineralizing medium. BD releases substances that can significantly induce formation of the human dental pulp stem cell-mineralized extracellular matrix, with physicochemical characteristics that are more conducive to pulp repair than those of MTA and CHC.

Short-pulse Er:YAG laser increases bond strength of composite resin to sound and eroded dentin.

This study evaluated the influence of the irradiation with a short-pulse Er:YAG laser on the adhesion of composite resin to sound and eroded dentin (SD and ED). Forty-six samples of occlusal dentine, obtained from human molars, had half of their surface protected, while the other half was submitted to erosive cycles. Afterward, 23 samples were irradiated with Er:YAG laser, resulting in four experimental groups: SD, sound irradiated dentine (SID­Er:YAG, 50 µs 50 µs , 2 Hz, 80 mJ, and 12.6 J/cm 2 12.6 J/cm2 ), ED, and eroded irradiated dentin (EID­erosion + Er:YAG laser). A self-etching adhesive system was used, and then cylinders of composite resin were prepared. A microshear bond strength test was performed after 24 h storage (n=20 n=20 ). The morphology of SD and ED, with or without Er:YAG laser irradiation, was evaluated under scanning electron microscopy (n=3 n=3 ). Bond strength values (MPa) were subjected to analysis of variance followed by Tukey's test. Statistically significant differences were found among the experimental groups: SD (9.76±3.39 B 9.76±3.39 B ), SID (12.77±5.09 A 12.77±5.09 A ), ED (5.12±1.72 D 5.12±1.72 D ), and EID (7.62±3.39 C 7.62±3.39 C ). Even though erosion reduces the adhesion to dentin, the surface irradiation with a short-pulse Er:YAG laser increases adhesion to both ED and SD.

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 signifi cativamente 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.

Análise in vitro da proliferação, diferenciação e migração de células-tronco de polpa dentária humana em resposta a materiais com potencial para serem empregados como capeadores pulpares diretos (óleo-resina de copaíba isolado ou associado a hidróxido de cálcio ou a agregado de trióxido mineral) / In vitro analysis of proliferation, differentiation and migration of human dental pulp stem cells in response to potential materials for direct pulp capping (oil-resin copaiba alone or associated to calcium hydroxyde or mineral trioxide aggregate)

O hidróxido de cálcio (HCa), o agregado de trióxido mineral (MTA) e o óleo-resina de copaíba (COP) isoladamente apresentam características biológicas do material de capeamento pulpar direto mais apropriado. Com o pressuposto que associados poderiam originar materiais mais apropriados para serem aplicados em capeamento pulpar direto, este estudo objetivou analisar in vitro proliferação, diferenciação e migração de células-tronco de polpa de dente decíduo humano esfoliado (SHEDs) em resposta a substâncias liberadas pelo COP isolado ou associado ao HCa ou ao MTA. Proliferação, diferenciação e migração de SHEDs (Linhagem PDH3) foram analisadas através do ensaio de redução do MTT; da atividade de fosfatase alcalina (ALP), formação de nódulos mineralizados pelo ensaio de Vermelho de Alizarina e expressão dos genes (BGLAP, DSPP, DMP1 e HSP-27) pelo qRT-PCR; e, do ensaio do Scratch, respectivamente. As células foram submetidas à ação de meios condicionados pelos biomateriais, de acordo com os seguintes grupos experimentais: COP isolado (COP); HCa isolado (HCa); HCa associado ao COP (HCa+COP); MTA isolado (MTA) e MTA associado ao COP (MTA+COP). Células crescidas em meio de cultura fresco serviram de controle. Os dados foram comparados utilizando ANOVA complementado pelo teste de Tukey (p 0,05). O grupo HCa apresentou número de células viáveis significativamente menor que o dos demais grupos, inclusive o do grupo HCa+COP (p<0,01) em todos os tempos experimentais...

The calcium hydroxide (CaH), the mineral trioxide aggregate (MTA) and the oil-resin copaiba (COP) by themselves have biological characteristics of the ideal direct pulp capping material. With the assumption that when associated they could originate materials more appropriated for direct pulp capping, this study aimed to analyze the in vitro proliferation, differentiation and migration of stem cells from human exfoliated deciduous teeth (SHEDs) in response to substances leached from COP alone or associated with CaH or MTA. Proliferation, differentiation and migration of SHEDs (PDH3 lineage) were analyzed through the MTT reduction assay; alkaline phosphatase (ALP) activity, mineralized nodules formation using the Alizarin Red assay and gene expression (BGLAP, DSPP, DMP1 e HSP-27) using qRT-PCR; and the Scratch assay, respectively. The cells were submitted to the culture medium conditioned by the biomaterials according to the following experimental groups: COP alone (COP); CaH alone (CaH); CaH associated to COP (CaH+COP); MTA alone (MTA) and MTA associated to COP (MTA+COP). Cells grown in fresh culture medium served as control. Data were compared by ANOVA complemented by the Tukey´s test (p 0.05). The CaH group presented number of viable cells significantly smaller (p<0.01) than those of all other experimental groups, including the CaH+COP, during whole experimental time. The ALP activity in 14 days was similar in all experimental groups. In 21 days, the COP group presented the amount of mineralization higher (p<0.01) than those of all other groups. The gene DMP1 was not expressed by the cells in all experimental groups. The COP group presented the smallest expression (p<0.01) of BGLAP, DSPP and HSP-27 genes. The SHEDs of the MTA+COP group presented superexpression of the BGLAP, DSPP and HSP-27 genes, and in the CaH+COP group the cells superexpressed the BGLAP and HSP-27 genes...