Influência da funcionalização do biovidro, preparado pela rota sonoquímica, com o fármaco teriparatida 10 na resposta celular in vitro e in vivo
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Data
2021-04-06
Autores
Souto, Juliani Caroline Ribeiro de Araújo
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Universidade Estadual Paulista (Unesp)
Resumo
O objetivo neste estudo foi avaliar a influência do biovidro 45S5 funcionalizado com teriparatida 10% na diferenciação e atividade de células mesenquimais, e no reparo ósseo em defeitos críticos realizados em ratas ovariectomizadas. O biomaterial foi produzido e caracterizado antes e após a funcionalização. Para o estudo in vitro, foram diferenciados osteoblastos obtidos a partir de células mesenquimais, isoladas de fêmures de ratas ovariectomizadas. Após diferenciação, as células foram submetidas ao MEV, MTT, conteúdo de proteína total, atividade de fosfatase alcalina (ALP), formação de nódulos de mineralização. No estudo in vivo, foram utilizadas 40 ratas Wistar, as quais foram divididas em dois grupos (n=20), grupo submetido à ovariectomia bilateral (OVX) e grupo submetido à cirurgia simulada de ovariectomia (Sham). Após 60 dias destes procedimentos, independente do grupo, todas as ratas foram submetidas à confecção de defeitos ósseos críticos de 5,0 mm, na calvária. No lado direito foram preenchidos com coágulo (controle) ,enquanto do lado esquerdo metade dos animais receberam preenchimento no defeito ósseo de biovidro (BG) e a outra metade recebeu como material de preenchimento o biovidro funcionalizado com teriparatida 10% (BGT). Após 2 e 6 semanas, os animais foram eutanasiados (n=10). A caracterização por MEV, espectroscopia de energia dispersiva (EDS), espectroscopia de infravermelho por transformação de Fourier (FTIR) e o potencial Zeta demonstraram que as amostras após a funcionalização apresentaram características morfológicas topográficas e químicas modificadas, indicativas de que a superfície foi alterada pelo processo de funcionalização. Na análise in vitro, observou que os grupos experimentais não foram citotóxicos, propiciaram um ambiente adequado à atividade e diferenciação celular e permitiram o espraiamento celular sobre as amostras. Na análise histológica descritiva os grupos experimentais, demonstraram neoformação óssea na região do defeito em ambos grupos e períodos. Na análise histomorfométrica, somente o grupo biovidro funcionalizado com o fármaco no grupo OVX no período de 6 semanas diferiu estatisticamente dos demais grupos (p<0,05) no período de 2 semanas. Conclui-se que estudos de longo prazo devem ser realizados para fornecer informações adicionais sobre o desempenho biológico da ação sinérgica entre os biovidros e a liberação do fármaco teriparatida.
The objective of this study was to evaluate the influence of the 45S5 bioglass functionalized with 10% teriparatide on the differentiation and activity of mesenchymal cells, and on bone repair in critical defects performed in ovariectomized rats. First, the biomaterial was produced and characterized before and after functionalization. For the in vitro study, osteoblasts obtained from mesenchymal cells, isolated from femurs of ovariectomized rats, were differentiated. After differentiation, the cells were submitted to SEM, MTT, total protein content, alkaline phosphatase activity (ALP), formation of mineralization nodules. In the in vivo study, 40 Wistar rats were used, which were initially divided into two groups (n = 20), a group undergoing bilateral ovariectomy (OVX) and a group undergoing simulated ovariectomy surgery (Sham). After 60 days of these procedures, regardless of the group, all rats were subjected to the making of 5.0 mm critical bone defects at calvaria, which were filled on the right side with a clot (control) and on the left side half of the animals received filling in the bone defect of bioglass (BG) and the other half received as functional filler the bioglass functionalized with 10% teriparatide (BGT). After 2 and 6 weeks, the animals were euthanized. SEM characterization, dispersive energy spectroscopy (EDS), Fourier transformation infrared spectroscopy (FTIR) and analysis of the Zeta potential demonstrated that the samples after functionalization showed modified topographic and chemical morphological characteristics, indicating that the surface was chemically altered by the functionalization process. The experimental groups were not cytotoxic and provided an adequate environment for cell adhesion and differentiation. Additionally, the analysis performed by SEM showed that all samples allowed cell spreading. In the descriptive histological analysis, the experimental groups demonstrated characteristics of bone neoformation with the presence of bone tissue in both periods. In the histomorphometric analysis, only the bioglass group functionalized with the drug in the OVX group differed statistically from the other groups (p <0.05) in both periods. It is concluded that long-term studies must be carried out to provide additional information on the biological performance of the synergistic action between bio-glasses and the release of the drug teriparatide.Concluded that long-term studies should be carried out to provide additional information on the mechanisms necessary to evaluate the biological performance of the synergistic action between bioglasses and the release of the drug teriparatide.
The objective of this study was to evaluate the influence of the 45S5 bioglass functionalized with 10% teriparatide on the differentiation and activity of mesenchymal cells, and on bone repair in critical defects performed in ovariectomized rats. First, the biomaterial was produced and characterized before and after functionalization. For the in vitro study, osteoblasts obtained from mesenchymal cells, isolated from femurs of ovariectomized rats, were differentiated. After differentiation, the cells were submitted to SEM, MTT, total protein content, alkaline phosphatase activity (ALP), formation of mineralization nodules. In the in vivo study, 40 Wistar rats were used, which were initially divided into two groups (n = 20), a group undergoing bilateral ovariectomy (OVX) and a group undergoing simulated ovariectomy surgery (Sham). After 60 days of these procedures, regardless of the group, all rats were subjected to the making of 5.0 mm critical bone defects at calvaria, which were filled on the right side with a clot (control) and on the left side half of the animals received filling in the bone defect of bioglass (BG) and the other half received as functional filler the bioglass functionalized with 10% teriparatide (BGT). After 2 and 6 weeks, the animals were euthanized. SEM characterization, dispersive energy spectroscopy (EDS), Fourier transformation infrared spectroscopy (FTIR) and analysis of the Zeta potential demonstrated that the samples after functionalization showed modified topographic and chemical morphological characteristics, indicating that the surface was chemically altered by the functionalization process. The experimental groups were not cytotoxic and provided an adequate environment for cell adhesion and differentiation. Additionally, the analysis performed by SEM showed that all samples allowed cell spreading. In the descriptive histological analysis, the experimental groups demonstrated characteristics of bone neoformation with the presence of bone tissue in both periods. In the histomorphometric analysis, only the bioglass group functionalized with the drug in the OVX group differed statistically from the other groups (p <0.05) in both periods. It is concluded that long-term studies must be carried out to provide additional information on the biological performance of the synergistic action between bio-glasses and the release of the drug teriparatide.Concluded that long-term studies should be carried out to provide additional information on the mechanisms necessary to evaluate the biological performance of the synergistic action between bioglasses and the release of the drug teriparatide.
Descrição
Palavras-chave
Diferenciação celular, Materiais biocompatíveis, Regeneração óssea, Osteoporose, Teriparatida, Biocompatible materials, Bone regeneration, Cell differentiation, Osteoporosis, Teriparatide