Efeito da aplicação local do curcumin nanoparticulado sobre o reparo ósseo in vivo e potencial osteogênico in vitro
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Data
2020-03-20
Autores
Silva, Amanda Favoreto
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Curcumin é um composto ativo derivado da planta Curcuma longa que exibe uma variedade de atividades biológicas, potencial anti-inflamatório e imunomodulatório, que pode ser associado ao tratamento de uma série de condições, como doenças inflamatórias intestinais, artrite reumatoide, câncer, diabetes e periodontite. Recentemente, a literatura também tem demonstrado o potencial do composto sobre o reparo ósseo, entretanto, os dados ainda são contraditórios. O objetivo deste estudo é avaliar o efeito da administração tópica do curcumin veiculado em nanopartículas de PLGA sobre a formação/reparo ósseo de defeitos críticos in vivo, e investigar seu potencial biológico in vivo e in vitro. Defeitos ósseos confeccionados na calvária de ratos machos foram preenchidos com solução salina (controle), curcumin nanoparticulado 0,05mg/ml ou veículo utilizado na diluição do composto (nanopartícula vazia) por 7, 14 e 28 dias. A fração de volume ósseo no interior do defeito nos diferentes grupos foram avaliados por microtomografia óssea (μCT); expressão e imunolocalização dos marcadores do turnover tecidual (RANKL, OPG, PCNA e RUNX2) foram investigados por imuno-histoquímica, enquanto as características teciduais do tecido neoformado foram observadas através da análise histológica descritiva e estereometria. In vitro, o potencial do curcumin nanoparticulado em estimular a diferenciação osteogênica de células estromais derivadas da medula óssea (BMSCs) de ratos foi avaliado determinando-se a atividade da fosfatase alcalina (ALP Assay) e produção de matriz mineralizada. uCT e análise histológica mostraram maior formação óssea nos defeitos de animais tratados curcumin no período de 14 dias. Curcumin reduziu o infiltrado celular e aumentou o conteúdo colágeno no período mais tardio (28 dias), e aumentou a expressão de PCNA e RUNX2 aos 14 dias. In vitro, a menor concentração de composto(0,025 µg /ml) aumentou significativamente a atividade de ALP em meio osteogênico, entretanto, não estimulou a produção de matriz mineralizada. Os resultados obtidos permitem concluir que o curcumin pode favorecer o reparo ósseo em defeitos críticos e estimular a osteoblastogênese de maneira dose-dependente in vitro.
Curcumin is an active compound derived from the Curcuma longa plant that exhibits a variety of biological activities, anti-inflammatory and immunomodulatory potential, which can be associated with the treatment of a number of conditions, such as inflammatory bowel diseases, rheumatoid arthritis, cancer, diabetes and periodontitis. Recently, the literature has also demonstrated the potential of the compound on bone repair, however, the data are still contradictory. The aim of this study is to evaluate the effect of topical administration of curcumin-loaded PLGA nanoparticles on bone formation / repair of critical defects in vivo, and to investigate its biological potential in vivo and in vitro. Bone defects made in the calvaria of rats were filled with saline solution (control), nanoparticulate curcumin 0,05mg/ml, or vehicle used in the dilution of the compound (empty nanoparticle) for 7, 14 and 28 days. The fraction of bone volume within the defect in the different groups was assessed by microcomputed tomography (μCT); expression and immunolocation of tissue turnover markers (RANKL, OPG, PCNA and RUNX2) were investigated by immunohistochemistry, while the tissue characteristics of the newly formed tissue were observed through descriptive histological analysis and stereometry. In vitro, the potential of nanoparticulate curcumin in stimulating osteogenic differentiation of rat bone marrow-derived stromal cells (BMSCs) was assessed by determining the activity of alkaline phosphatase (ALP Assay) and mineralized matrix production. uCT and histological analysis showed greater bone formation in the defects of animals treated curcumin in the period of 14 days. Curcumin reduced cell infiltrate and increased collagen content in the later period (28 days), and increased expression of PCNA and RUNX2 at 14 days. In vitro, the lower concentration of compound (0,025 µg /ml) significantly increased the activity of ALP in osteogenic medium, however, it did not stimulate the production of mineralized matrix. The results obtained allow us to conclude that curcumin can favor bone repair in critical defects and stimulate osteoblastogenesis in a dose-dependent manner in vitro.
Curcumin is an active compound derived from the Curcuma longa plant that exhibits a variety of biological activities, anti-inflammatory and immunomodulatory potential, which can be associated with the treatment of a number of conditions, such as inflammatory bowel diseases, rheumatoid arthritis, cancer, diabetes and periodontitis. Recently, the literature has also demonstrated the potential of the compound on bone repair, however, the data are still contradictory. The aim of this study is to evaluate the effect of topical administration of curcumin-loaded PLGA nanoparticles on bone formation / repair of critical defects in vivo, and to investigate its biological potential in vivo and in vitro. Bone defects made in the calvaria of rats were filled with saline solution (control), nanoparticulate curcumin 0,05mg/ml, or vehicle used in the dilution of the compound (empty nanoparticle) for 7, 14 and 28 days. The fraction of bone volume within the defect in the different groups was assessed by microcomputed tomography (μCT); expression and immunolocation of tissue turnover markers (RANKL, OPG, PCNA and RUNX2) were investigated by immunohistochemistry, while the tissue characteristics of the newly formed tissue were observed through descriptive histological analysis and stereometry. In vitro, the potential of nanoparticulate curcumin in stimulating osteogenic differentiation of rat bone marrow-derived stromal cells (BMSCs) was assessed by determining the activity of alkaline phosphatase (ALP Assay) and mineralized matrix production. uCT and histological analysis showed greater bone formation in the defects of animals treated curcumin in the period of 14 days. Curcumin reduced cell infiltrate and increased collagen content in the later period (28 days), and increased expression of PCNA and RUNX2 at 14 days. In vitro, the lower concentration of compound (0,025 µg /ml) significantly increased the activity of ALP in osteogenic medium, however, it did not stimulate the production of mineralized matrix. The results obtained allow us to conclude that curcumin can favor bone repair in critical defects and stimulate osteoblastogenesis in a dose-dependent manner in vitro.
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Palavras-chave
Curcumin, Células-Tronco Mesenquimais, Nanopartículas, Ratos, Regeneração óssea, Bone marrow stromal cells, Nanoparticles, Rats, Bone Regeneration