Avaliação in vitro de sistemas adesivos de dentina aplicados sobre células odontoblastóides
In vitro evaluation of adhesive systems applied on culture of odontoblas-like cells
Costa, C.A.S.; Hebling, J.
Rev. odontol. UNESP, vol.35, n1, p.97-106, 2006
Resumo
Objetivo: Avaliar a citotoxicidade de dois agentes adesivos dentinários. Material e método: Células imortalizadas de linhagem odontoblástica (MDPC-23) foram semeadas (30.000 células/cm2) sobre lamínulas de vidro posicionadas na base de 4 recipientes com 24 compartimentos. Após 24 horas de incubação, o número de células aderidas ao substrato foi contado. Sobre essas células, foram colocados 995 μL de meio de cultura, no qual era adicionado 5 μL de cada material experimental: Grupo 1- Clearfil Liner Bond 2V (Kuraray Co.); Grupo 2- Single Bond (3M ESPE); e Grupo 3- Hidróxido de cálcio (Pathfinder Associates Inc.). No Grupo 4 (controle), 5 μL de tampão fosfato foi adicionado ao meio de cultura, e o número de células foi novamente contado. O metabolismo celular foi avaliado pela técnica do methyltetrazolium, sendo a morfologia das células caracterizada em MEV. Os dados numéricos obtidos foram submetidos à análise estatística de ANOVA complementada pelos testes de Tukey, t-student, e Dunnett. Resultados: Para os grupos 1, 2 e 3, o número de células reduziu em 65, 73 e 14%, respectivamente. Os testes estatísticos determinaram a significante diferença entre todos os grupos comparados entre si ou ao grupo controle. A redução no metabolismo celular para os grupos 1, 2 e 3 foi de 99, 99 e 37%, respectivamente. Conclusão: Ambos os agentes adesivos experimentais causaram intenso efeito citotóxico sobre as células odontoblastóides, sendo o hidróxido de cálcio considerado o material menos citopático para as células MDPC-23.
Palavras-chave
Citotoxicidade, sistemas adesivos, cultura de células, odontoblastos
Abstract
To evaluate the cytotoxicity of two current dentin bonding agents. Material and methods: The immortalized odontoblast cell line (MDPC-23) was plated (30,000 cell/cm2) on round glass discs placed in the bottom of 24 well dishes. After 72 hours incubation the cell number was counted. The following dental materials (5 μL) were added to the culture medium (995 μL) and applied for 120 minutes to the cultured cells in 24 wells for each group: Group 1- Clearfil Liner Bond 2V (Kuraray Co.); Group 2- Single Bond (3M ESPE); and Group 3- Calcium hydroxide (Pathfinder Associates Inc.). In group 4 (control), phosphate buffer saline solution was added to the fresh medium. The cell number was counted again. The cytotoxic effects were evaluated by using the MTT assay and the cell morphology was analysed by SEM. The scores obtained from the cell count and MTT assays were statistically analyzed by the ANOVA method complemented by Tukey, t-student, and Dunnett tests. Results: In groups 1, 2 and 3 the experimental materials decreased the cell number by 65, 73, and 14%, respectively. The difference among the experimental and control groups was statistically significant. In groups 1, 2, and 3, the materials reduced the cell metabolism by 99, 99, and 37%, respectively. Conclusion: Both experimental adhesive systems caused very high cytophatic effects to the MDPC-23 cells. On the other hand, the calcium hydroxide was the least cytotoxic material evaluated.
Keywords
Cytotoxicity, adhesive systems, culture of cells, odontoblasts
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