Revista de Odontologia da UNESP
http://revodontolunesp.com.br/article/doi/10.1590/1807-2577.1087
Revista de Odontologia da UNESP
Original Article

Biological characterization of implant surfaces - in vitro study

Caracterização biológica de superfície de titânio - estudo in vitro

Soares, Priscilla Barbosa Ferreira; Moura, Camilla Christian Gomes; Rocha Júnior, Huberth Alexandre da; Dechichi, Paula; Zanetta-Barbosa, Darceny

Abstract

Objective: Evaluate the biological performance of titanium alloys grade IV under different surface treatments: sandblasting and double etching (Experimental surface 1; Exp1, NEODENT); surface with wettability increase (Experimental surface 2; Exp2, NEODENT) on response of preliminary differentiation and cell maturation. Material and method: Immortalized osteoblast cells were plated on Exp1 and Exp2 titanium discs. The polystyrene plate surface without disc was used as control group (C). Cell viability was assessed by measuring mitochondrial activity (MTT) at 4 and 24 h (n = 5), cell attachment was performed using trypan blue exclusion within 4 hours (n = 5), serum total protein and alkaline phosphatase normalization was performed at 4, 7 and 14 days (n = 5). Data were analyzed using one-way ANOVA and Tukey test. Result: The values of cell viability were: 4h: C– 0.32±0.01A; Exp1– 0.34±0.08A; Exp2– 0.29±0.03A. 24h: C– 0.43±0.02A; Exp1– 0.39±0.01A; Exp2– 0.37±0.03A. The cell adhesion counting was: C– 85±10A; Exp1- 35±5B; Exp2– 20±2B. The amounts of serum total protein were 4d: C– 40±2B; Exp1– 120±10A; Exp2– 130±20A. 7d: C– 38±2B; Exp1– 75±4A; Exp2– 70±6A. 14 d: C– 100±3A; Exp1– 130±5A; Exp2– 137±9A. The values of alkaline phosphatase normalization were: 4d: C– 2.0±0.1C; Exp1– 5.1±0.8B; Exp2– 9.8±2.0A. 7d: C– 1.0±0.01C; Exp1– 5.3±0.5A; Exp2– 3.0±0.3B. 14 d: C– 4.1±0.3A; Exp1– 4.4±0.8A; Exp2– 2.2±0.2B. Different letters related to statistical differences. Conclusion: The surfaces tested exhibit different behavior at dosage of alkaline phosphatase normalization showing that the Exp2 is more associated with induction of cell differentiation process and that Exp1 is more related to the mineralization process.

Keywords

Titanium surface, mitochondrial colorimetric assay, cell cytotoxicity.

Resumo

Objetivo: Avaliar o desempenho biológico de ligas de titânio grau IV submetidos a diferentes tratamentos de superfície – jateamento e duplo ataque ácido (Superfície experimental 1; Exp1, NEODENT) e superfície com aumento na molhabilidade (Superfície experimental 2; Exp2, NEODENT) em resposta preliminar de diferenciação e maturação celular. Material e método: Foram plaqueados osteoblastos imortalizados sobre discos de titânio de Exp1 e Exp2 e como controle o poço da placa de cultura sem disco (C). Empregou-se ensaios de viabilidade celular (MTT) em 4 e 24 horas (n = 5), adesão celular em 4 horas (n = 5), dosagem de proteínas totais e fosfatase alcalina normalizada em 4, 7 e 14 dias (n = 5). Os dados foram analisados por ANOVA em fator único seguido de teste de Tukey. Resultado: Os valores de viabilidade celular foram: 4h: C– 0,32±0,01A; Exp1 – 0,34±0,08A; Exp2– 0,29±0,03A. 24h: C– 0,43±0,02A; Exp1– 0,39±0,01A; Exp2– 0,37±0,03A. A contagem de adesão celular foi: C– 85±10A; Exp1– 35±5B; Exp2– 20±2B. Os valores de proteínas totais foram: 4d: C– 40±2B; Exp1– 120±10A; Exp2– 130±20A. 7d: C– 38±2B; Exp1– 75±4A; Exp2– 70±6A. 14 d: C– 100±3A; Exp1– 130±5A; Exp2– 137±9A. Os valores de fosfatase alcalina normalizada foram: 4d: C– 2,0±0,1C; Exp1– 5,1±0,8B; Exp2– 9,8±2,0A, 7d: C– 1,0±0,01C; Exp1– 5,3±0,5A; Exp2– 3,0±0,3B, 14 d: C– 4,1±0,3A; Exp1– 4,4±0,8A; Exp2– 2,2±0,2B. Letras diferentes representam diferenças estatísticas. Conclusão: As superfícies testadas apresentaram comportamento diferenciado na dosagem de fosfatase alcalina normalizada traduzindo que Exp2 está relacionado com processo de indução de diferenciação celular e Exp1 relacionado com processo de mineralização.

Palavras-chave

Superfície de titânio, ensaio colorimétrico mitocondrial, citotoxidade celular.

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