Efeito do substituto ósseo particulado associado ou não ao MTA na citotoxicidade, resposta tecidual e reparo ósseo em defeitos críticos em calvária de ratos
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Data
2019-08-29
Autores
Machado, Thiago [UNESP]
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
A utilização de biomateriais que visam devolver o volume ósseo perdido após a perda dentária tem se expandido. O osso é um tecido conjuntivo altamente especializado, possuindo dinâmica aposicional onde o equilíbrio entre neoformação e reabsorção envolve a interação de fatores endócrinos, parácrinos e autócrinos. Diversas associações de materiais diversos e substitutos ósseos têm sido estudadas, porém, o MTA (Agregado Trióxido Mineral) ainda carece de informações acerca da sua utilização como substituto ósseo ou em associação com demais substitutos. Assim, o objetivo deste estudo foi avaliar a influência da presença do MTA Angelus Branco® nas proprorções de 5%, 10% e 15% em associação com substituto ósseo de Hidroxiapatita e β- Tricálcio Fosfato na citotoxicidade, resposta tecidual e reparo ósseo, em defeito crítico em calvária de ratos. Para tanto, utilizou-se cultura celular da SAOS-2 para avaliação citológica e ensaio MTT do contato direto e eluentes. Também foram utilizados 112 ratos machos Wistar distribuídos em 7 grupos e avaliados em 2 tempos (7 e 28 dias). Após eutanasiados foram submetidos à microtomografia e por coloração de hematoxilina e eosina para análise histológica e histomorfométrica. A análise da homocedasticidade foi realizada pelo teste Shapiro-Wilk, para distinção dos dados paramétricos e não paramétricos. Para análise dos dados paramétricos, foi realizada análise de variância One-Way (ANOVA One-Way) e realizado o pós-teste de Tukey para os parâmetros microtomográficos e para a citotoxicidade foi aplicado o teste de correção de Bonferroni. Para os dados não paramétricos utilizou-se o Kruskal-Wallis e Mann Whitney. Adotou-se o nível de significância de p <0,05. Os parâmetros volume ósseo (BV), porcentagem de volume ósseo (BV/TV), espessura do trabeculado ósseo (Tb.Th) e número de trabéculas (Tb.N) não apresentaram diferenças estatísticas. Sendo o grupo Osteosynt® +15% MTA o grupo com maiores valores médios para os parâmetros BV, BV/TV, Tb.Th e Tb.N. A análise histomorfométrica para neoformação óssea e reabsorção dos biomateriais não apresentaram diferenças estatísticas. De acordo com os resultados, pode-se concluir que, estatisticamente, a adição de MTA nas 3 proporções testadas não demonstrou efetividade quando comparada ao uso isolado dos substitutos ósseos Bio-Oss® e Osteosynt® e coágulo. No entanto, foi evidenciada tendências à superioridade numérica quanto ao volume ósseo neoformado da associação Osteosynt® + 15% MTA aos 7 dias e Osteosynt® + 5% MTA e Osteosynt® + 10% MTA nas análises aos 28 dias.
The use of biomaterials that aim to return lost bone volume after tooth loss has expanded. Bone is a highly specialized connective tissue with appositional dynamics where the balance between neoformation and resorption involves the interaction of endocrine, paracrine and autocrine factors. Several associations of different materials and bone substitutes have been studied. However, the MTA (Mineral Trioxide Aggregate) still lacks information about its use as bone substitute or in association with other substitutes. Thus, the aim of this study was to evaluate the influence of presence of MTA Angelus Branco® in the 5%, 10% and 15% proportions in association with Hydroxyapatite and β-Tricalcium Phosphate bone substitute in cytotoxicity, tissue response and bone repair in critical defect in rat calvaria. For this, SAOS-2 cell culture was used for cytological evaluation and MTT assay of direct contact and eluents. We also used 112 male Wistar rats distributed into 7 groups and evaluated at 2 distinct times (7 and 28 days). After euthanasia, the specimens were submitted to microtomography and hematoxylin and eosin staining for histological and histomorphometric analysis. The analysis of homoscedasticity was performed by the Shapiro-Wilk test for distinction of parametric and non-parametric data. For analysis of parametric data, one-way analysis of variance (One-Way ANOVA) was performed and Tukey's post-test was performed for the microtomographic parameters and for the cytotoxicity the Bonferroni correction test was applied. For non-parametric data we used the Kruskal-Wallis and Mann Whitney. The significance level of p <0.05 was adopted. The parameters bone volume (BV), bone volume percentage (BV / TV), bone trabecular thickness (Tb.Th) and number of trabeculae (Tb.N) showed no statistical differences. The Osteosynt® + 15% MTA group was the group with the highest mean values for the parameters BV, BV / TV, Tb.Th and Tb.N. Histomorphometric analysis for bone neoformation and biomaterial resorption showed no statistical differences. According to the results, it can be concluded that, statistically, the addition of MTA in the 3 tested proportions did not show effectiveness when compared to the isolated use of the Bio-Oss® and Osteosynt® bone substitutes and clot. However, trends in numerical superiority regarding the newly formed bone volume of the association Osteosynt® + 15% MTA at 7 days and Osteosynt® + 5% MTA and Osteosynt® + 10% MTA in the analyzes at 28 days were evidenced.
The use of biomaterials that aim to return lost bone volume after tooth loss has expanded. Bone is a highly specialized connective tissue with appositional dynamics where the balance between neoformation and resorption involves the interaction of endocrine, paracrine and autocrine factors. Several associations of different materials and bone substitutes have been studied. However, the MTA (Mineral Trioxide Aggregate) still lacks information about its use as bone substitute or in association with other substitutes. Thus, the aim of this study was to evaluate the influence of presence of MTA Angelus Branco® in the 5%, 10% and 15% proportions in association with Hydroxyapatite and β-Tricalcium Phosphate bone substitute in cytotoxicity, tissue response and bone repair in critical defect in rat calvaria. For this, SAOS-2 cell culture was used for cytological evaluation and MTT assay of direct contact and eluents. We also used 112 male Wistar rats distributed into 7 groups and evaluated at 2 distinct times (7 and 28 days). After euthanasia, the specimens were submitted to microtomography and hematoxylin and eosin staining for histological and histomorphometric analysis. The analysis of homoscedasticity was performed by the Shapiro-Wilk test for distinction of parametric and non-parametric data. For analysis of parametric data, one-way analysis of variance (One-Way ANOVA) was performed and Tukey's post-test was performed for the microtomographic parameters and for the cytotoxicity the Bonferroni correction test was applied. For non-parametric data we used the Kruskal-Wallis and Mann Whitney. The significance level of p <0.05 was adopted. The parameters bone volume (BV), bone volume percentage (BV / TV), bone trabecular thickness (Tb.Th) and number of trabeculae (Tb.N) showed no statistical differences. The Osteosynt® + 15% MTA group was the group with the highest mean values for the parameters BV, BV / TV, Tb.Th and Tb.N. Histomorphometric analysis for bone neoformation and biomaterial resorption showed no statistical differences. According to the results, it can be concluded that, statistically, the addition of MTA in the 3 tested proportions did not show effectiveness when compared to the isolated use of the Bio-Oss® and Osteosynt® bone substitutes and clot. However, trends in numerical superiority regarding the newly formed bone volume of the association Osteosynt® + 15% MTA at 7 days and Osteosynt® + 5% MTA and Osteosynt® + 10% MTA in the analyzes at 28 days were evidenced.
Descrição
Palavras-chave
Substitutos ósseos, Regeneração óssea, Sobrevivência celular, Ratos, Bone graft substitutes, Bone regeneration, Cell survival, Rats