Biological effect of bioactive cements on stem cells from human exfoliated deciduous teeth

Author(s)

Ana Beatriz Vieira da Silveira

• Document type: Master's Dissertation
• Press: Bauru.
• Institution: Universidade de São Paulo (USP). Faculdade de Odontologia de Bauru (FOB/SDB)
• Defense date: 2023-10-06

Examining board members

Thais Marchini de Oliveira Valarelli; Maria Aparecida de Andrade Moreira Machado; Raquel Assed Bezerra Segato; Juliane Maria Guerreiro Tanomaru

• Advisor: Thais Marchini de Oliveira Valarelli

Abstract

This dissertation had the purpose of presenting three scientific articles. Article 1: The aim of the present study was to evaluate the cytotoxicity of extracts of bioceramic materials in different dilutions. Stem cells from exfoliated human deciduous teeth (SHED) were used. The samples were immersed in MEM 10% FCS according to the groups: BBio Membrane, Bio-C Repair, MTA HP Repair, TheraCal LC and Biodentine. The methods used were MTT and Alamar Blue, at 24, 48 and 72 hours after contact with the bioceramic materials at 1:1 and 1:2 dilutions. Data were analyzed using the three-way ANOVA test, followed by the Tukey test (p<0.05). The results showed that BBio Membrane, Bio-C Repair and Biodentine extracts in the 1:1 dilution were more cytotoxic when compared to the 1:2 dilution in all studied periods. It is concluded that the cytotoxicity of the bioceramic material is related to the dilution of the extract used. MTA HP Repair showed high biocompatibility, as even at a 1:1 dilution it was cytocompatible. The BBio Membrane proved to be a promising new biomaterial. Article 2: The purpose of this study was to analyze the biological effects of bioactive cements on SHED. SHED samples were obtained from a Biorepository. The extracts were prepared at a 1:2 dilution, as follows: Bio-C Repair (G1), MTA HP Repair (G2) and Biodentine (G3). Cell viability, migration, phosphatase activity and mineralization nodule tests were performed. Data were analyzed using the two-way ANOVA test, followed by the Tukey or Kruskal-Wallis test, followed by the Bonferroni test (p<0.05). It was observed that in cell viability Bio-C Repair and Biodentine presented superior results to MTA HP Repair. In cell migration, in 72H, cells treated with MTA HP Repair showed greater cleft closure. In alkaline phosphatase activity, SHED that came into contact with Bio-C Repair extract had higher ALP activity in 10 days. Biodentine had a significant stimulating effect on the formation of mineralized nodules. According to the results obtained, Bio-C Repair, MTA HP Repair and Biodentine highlight bioactive properties suitable for vital pulp therapy procedures in deciduous teeth. Article 3: The aim of this study was to evaluate the viability and gene expression of SHED for odontogenic/osteogenic and angiogenic differentiation after contact with different bioceramic materials. The extracts were divided into groups: Dentine/pulp membrane (BBio) (G1), Bio-C Repair (G2), MTA HP Repair (G3) and Biodentine (G4). Alamar Blue and RT-PCR tests were performed for the targets: COL, FGF-2, VEGF, OCN, OPN, DSPP and DMP-1. Data were analyzed using the two-way ANOVA test, followed by the Tukey test (p<0.05). The results showed that the bioceramic materials maintained the SHED viability, with emphasis on BBio Membrane and Bio-C Repair. Biodentine showed the best VEGF and FGF-2 expression results. The BBio Membrane stood out in the expression of OCN and OPN. Regarding DSPP and DMP-1, the cells showed a progressive increase in expression over time for all groups. It is concluded that all tested materials exhibited acceptable bioactivity. The BBio Membrane showed better capacity for osteo/odontogenic differentiation. (AU)

• FAPESP's process: 21/08730-4 - Biological effects of bioactive cement on stem cells from human exfoliated deciduous teeth
• Grantee: Ana Beatriz Vieira da Silveira
• Support Opportunities: Scholarships in Brazil - Master