Abstract Objective: To examine the cytotoxicity of calcium hydroxide on human umbilical cord mesenchymal stem cells (HUCMSC) to understand the characteristics for use in regenerative dentistry procedures especially regenerative endodontics. Material and Methods: HUCMSC was isolated, cultured, and confirmed by flow cytometry. The biological characteristics, such as cell morphology, proliferation, and protein expression, were screened. To check the cytotoxicity, HUCMSC was cultured and divided into two groups, the control group (cultured in minimum essential medium (MEM) alpha) and calcium hydroxide group (cultured in MEM alpha and calcium hydroxide). Methyl-thiazole-tetrazolium (MTT) assay was done on different concentrations of calcium hydroxide (0.39 to 25 µg/mL) and the cells were observed and counted. One-way ANOVA test was used with a significance level set at 5%. Results: Flow cytometric analysis confirmed positive of CD73, CD90, CD105, negative of CD45 and CD34. A significant difference was found between the concentration of 6.25 and 3.125 µg/mL (p=0.004). There was no significant difference among 6.25, 12.5 and 25 µg/mL concentrations. There was also no significant difference among 0.39, 0.78, 1.56, and 3.125 µg/mL concentrations. Conclusion: Even though calcium hydroxide is a medicament of choice in clinical endodontics, it decreases the viability of HUCMSC. The lower the concentration of calcium hydroxide, the higher the viability of HUCMSC.

Resumo Os principais materiais para adesão dentinária em restaurações de resina composta são o 2-hidroxietil metacrilato (HEMA). O HEMA possui propriedades físicas e químicas benéficas e estáveis, ainda que tóxicas. A adição de etanol ou água pode reduzir o efeito tóxico do HEMA. O solvente etanol possui uma menor capacidade de ligação H em comparação com o solvente água, de modo que pode ligar menos radicais livres do monômero residual. Este trabalho teve como objetivo analisar a apoptose pela aplicação de adesivos dentinários com solventes etanol e água. Células de cultura de fibroblastos foram obtidas a partir do terceiro molar extraído, por meio do método de tripsinaion. As células foram divididas em 4 grupos como confluentes: cultura celular sem tratamento como controle, cultura celular com arcabouço de quitosana, cultura celular com arcabouço e adesivo dentinário polimerizado com solvente etanol ou água. A observação da apoptose foi realizada utilizando o método imunohistoquímico com coloração com brometo de etídio e acridina laranja, sob microscópio de fluorescência com aumento de 40´. Houve uma diferença significativa entre os grupos (p = 0,0001), mas não houve diferenças entre os solventes. A taxa de apoptose em cultura de células de fibroblastos expostos à adesão baseada em HEMA com solvente etanol foi de 67%, enquanto as células expostas à adesão baseada em HEMA com solvente de água foi de 44%. O efeito da adesão dentinária com solvente etanol e solvente água sobre a taxa de apoptose de células de fibroblastos de polpa não é diferente.

Abstract The most common main materials for dentin bonding for composite resin restoration is 2-hydroxyethyl methacrylate (HEMA). HEMA has beneficial physical and chemical properties, and stable, yet toxic. The addition of ethanol or water, may reduce the toxic effect of HEMA. Ethanol solvent has lower H-bonding capacity compared to water solvent, so it can bind less free radicals from the residual monomer. This study aimed to analyze apoptosis due to dentine bonding application with ethanol and water solvent. Fibroblast culture cells were obtained from extracted third molar, by means of tripsinasion method. The cells were divided into 4 groups as reached confluent: cell culture without treatment as control, cell culture with scaffold chitosan, cell culture with scaffold and polymerized dentin bonding with ethanol or water solvent. Apoptosis observation was conducted using immunohistochemistry method with ethidium bromide acridin orange staining, under fluorescent microscope with 40´ magnification. There was a significant difference among groups (p=0.0001), yet no differences found between different solvent. Apoptosis rate in fibroblast cells culture exposed to HEMA bonding with ethanol solvent was 67%, while the cells exposed to HEMA bonding with water solvent was 44%. The effect of dentin bonding with ethanol solvent and water solvent towards apoptosis rate of pulp fibroblast cells is not different.