Avaliação do reparo ósseo de defeitos críticos tratados com biomateriais nanocompósitos de poliamida 6 e nanopartículas detrimetafosfato decoradas com nanopartículas de prata: estudo histomorfométrico, microtomográfico e imunoistoquímico em calvárias de ratos
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Data
2021-12-17
Autores
Morabito, Maria Juliana Sismeiro Dias [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O objetivo deste estudo foi avaliar a influência de materiais nanocompósitos, obtidos a partir da incorporação de nanopartículas de trimetafosfato de sódio (TMP) decoradas com nanopartículas de prata a 2,5% (AgNP) em matrizes poliméricas de poliamida-6 (Pa6), no reparo ósseo de defeitos de tamanho crítico (DTC) criados cirurgicamente em calvárias de ratos. Trinta ratos Wistar machos foram divididos em três grupos, de acordo com o tratamento: Grupo Controle (C) - coágulo sanguíneo; Pa6-AgNP; e Pa6-TMP-AgNP. Um DTC de 5 mm de diâmetro foi criado na calvária de cada animal. No Grupo C, o defeito foi preenchido com coágulo sanguíneo somente, e nos outros grupos com scaffolds com Pa6-AgNP e Pa6-TMP-AgNP. Os animais foram eutanasiados aos 30 dias pós- operatórios. As Análises microtomográfica, histomorfométrica eimunoistoquímica foram realizadas. Assim, a área de osso neoformado (AON) foi calculada como uma porcentagem da área total do defeito original, e imunomarcações para TGFβ-1, BMP-2/4 e OCN foram determinadas. A partir da análise intergrupos, observou-se que o Grupo Pa6-TMP-AgNP apresentou maior área óssea neoformada e, o Grupo Pa6-AgNP apresentou AON superior ao grupo controle (P <0,05). A imunohistoquímica revelou que os grupos Pa6-TMP-AgNP e Pa6-AgNP apresentaramalta e moderada imunomarcação para TGFβ-1 e OCN, respectivamente. O Pa6-TMP-AgNP apresentou um maior número de células BMP-2/4 positivas em comparação aos demais grupos (p <0,001). A partir desses resultados, podemos concluirque Pa6-TMP-AgNP proporciona resposta biológica de neoformação óssea, assimcomo comportamento osteoindutor e osteocondutor quando utilizado para preencher defeitos críticos criados em calvária de ratos.
The aim of this study was to evaluate the influence of nanocomposite materials obtained from the incorporation of sodium trimetaphosphate (TMP) nanoparticles decorated with 2.5% silver nanoparticles (AgNP) in polymeric matrices of polyamide-6 (Pa6), in bone repair of surgically created critical size defects (CDA) in rat calvaria. Thirty male Wistar rats were divided into three groups, according to treatment: Control Group (C) - blood clot; Pa6-AgNP; and Pa6-TMP-AgNP. A 5mm diameter DTC was created on the calvaria of each animal. In Group C, the defect was filled with blood clot only, and in the other groups with scaffolds with Pa6-AgNP and Pa6-TMP-AgNP. The animals were euthanized 30 days after surgery. Microtomographic, histomorphometric and immunohistochemical analyzes were performed. Thus, the area of neoformed bone (AON) was calculated as a percentage of the total area of the original defect, and immunostaining for TGFβ-1, BMP- 2/4 and OCN was determined. From the intergroup analysis, it was observed that the Pa6- TMP-AgNP Group had a larger neoformed bone area, and the Pa6-AgNP Group had higher AON than the control group (P < 0.05). Immunohistochemistry revealed that the Pa6-TMP-AgNP and Pa6-AgNP groups showed high and moderate immunostaining for TGFβ-1 and OCN, respectively. Pa6-TMP-AgNP had a higher number of BMP-2/4 positive cells compared to the other groups (p <0.001). From these results, we can conclude that Pa6-TMP-AgNP provides biological response to bone neoformation, as well as osteoinductive and osteoconductive behavior when used to fill critical defects created in rat calvaria.
The aim of this study was to evaluate the influence of nanocomposite materials obtained from the incorporation of sodium trimetaphosphate (TMP) nanoparticles decorated with 2.5% silver nanoparticles (AgNP) in polymeric matrices of polyamide-6 (Pa6), in bone repair of surgically created critical size defects (CDA) in rat calvaria. Thirty male Wistar rats were divided into three groups, according to treatment: Control Group (C) - blood clot; Pa6-AgNP; and Pa6-TMP-AgNP. A 5mm diameter DTC was created on the calvaria of each animal. In Group C, the defect was filled with blood clot only, and in the other groups with scaffolds with Pa6-AgNP and Pa6-TMP-AgNP. The animals were euthanized 30 days after surgery. Microtomographic, histomorphometric and immunohistochemical analyzes were performed. Thus, the area of neoformed bone (AON) was calculated as a percentage of the total area of the original defect, and immunostaining for TGFβ-1, BMP- 2/4 and OCN was determined. From the intergroup analysis, it was observed that the Pa6- TMP-AgNP Group had a larger neoformed bone area, and the Pa6-AgNP Group had higher AON than the control group (P < 0.05). Immunohistochemistry revealed that the Pa6-TMP-AgNP and Pa6-AgNP groups showed high and moderate immunostaining for TGFβ-1 and OCN, respectively. Pa6-TMP-AgNP had a higher number of BMP-2/4 positive cells compared to the other groups (p <0.001). From these results, we can conclude that Pa6-TMP-AgNP provides biological response to bone neoformation, as well as osteoinductive and osteoconductive behavior when used to fill critical defects created in rat calvaria.
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Palavras-chave
Nanocompostos, Fosfatos, Prata, Nanopartículas, Imuno-histoquímica, Phosphates