Efeitos de scaffolds desenvolvidos a partir de compósitos bioativos extraídos de esponjas marinhas no processo de reparo ósseo de ratas osteoporóticas
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Data
2023-12-20
Autores
Cruz, Matheus de Almeida [UNIFESP]
Orientadores
Renno, Ana Claudia Muniz [UNIFESP]
Tipo
Tese de doutorado
Título da Revista
ISSN da Revista
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Resumo
Durante o processo de reparo ósseo, cerca de 5 a 20% dos casos podem culminar em um processo de consolidação anormal e resultar em um atraso na consolidação ou não união óssea. Uma vez este quadro instalado, pode gerar prejuízos na qualidade de vida destes indivíduos, altos índices de morbimortalidade e elevados custos ao sistema de saúde. Desta forma, se faz necessário a investigação de tratamentos que apresentem potencial osteogênico e que tenham a capacidade de acelerar o processo de reparo ósseo. Dentre estes recursos, pode-se evidenciar a aplicação de biomateriais, por exemplo, os materiais bioativos, como o biovidro (BG) e dos provenientes de esponjas marinhas como a biosilica (BS) e a espongina (SPG). Assim, o objetivo do presente estudo foi avaliar a biocompatibilidade dos compósitos de BS e BG associados ainda à SPG, por meio de estudo in vitro e in vivo, utilizando um modelo de defeito ósseo em tíbias de ratas osteoporóticas. Para o estudo in vitro, células MC3T3-E1. L929 e CHOK-1 foram analisadas através da viabilidade celular, ensaio de cometa, teste de micronúcleo e vermelho de alizarina. No estudo in vivo foram realizadas as seguintes análises: Histopatológica, histomorfométrica, Análise de colágeno e Imunohistoquímica. O estudo in vitro demostrou que, após 7 dias de cultura celular, houve um aumento significativo de viabilidade celular do grupo controle em comparação com os grupos BS, BS/SPG, BG e BG/SPG na concentração de 100%, nenhum dos materiais apresentou sinais de Genotoxicidade e os materiais apresentaram marcação por vermelho de alizarina. Para análise histológica, in vivo, 15 dias após a cirurgia, os grupos apresentaram tecido de granulação e algumas áreas de tecido ósseo neoformado. Trinta dias após a cirurgia, os grupos revelaram um tecido ósseo neoformado mais maduro e progressiva degradação do material. Todos os grupos tratados apresentaram deposição de colágeno organizado em rede após 30 dias. A análise de imuno-histoquímica demonstrou uma maior imunomarcação para Runx-2 em todos os grupos que receberam implantes de biomaterial, 15 e 30 dias após a cirurgia. Para a análise de OPG, observou-se um aumento da Imunomarcação para BS/SPG e BG/SPG em comparação com CG e BS e BG, 30 dias após a cirurgia. Desta forma, pode-se concluir que os estudos in vitro e in vivo apontaram que as amostras de BS e BS/SPG promoveram maior viabilidade celular e melhores propriedades biológicas nos defeitos ósseos na tíbia de ratas osteoporóticas, destacando o potencial dos compósitos associados a SPG para serem usados como enxertos ósseos em aplicações na medicina regenerativa.
During the bone repair process, approximately 5 to 20% of cases may culminate in an abnormal consolidation process and result in a delay in bone union or non-union. Once this situation is established, it can lead to losses in the quality of life of these individuals, high rates of morbidity and mortality and high costs to the health system. Therefore, it is necessary to investigate treatments that have osteogenic potential and that have the ability to accelerate the bone repair process. Among these resources, the application of biomaterials can be highlighted, for example, bioactive materials, such as bioglass (BG) and those from marine sponges such as biosilica (BS) and spongin (SPG). Thus, the objective of the present study was to evaluate the biocompatibility of BS and BG composites associated with SPG, through in vitro and in vivo studies, using a bone defect model in the tibias of osteoporotic rats. For the in vitro study, MC3T3-E1 cells. L929 and CHOK-1 were analyzed using cell viability, comet assay, micronucleus test and alizarin red. In the in vivo study, the following analyzes were carried out: Histopathological, histomorphometric, collagen analysis and immunohistochemistry. The in vitro study demonstrated that, after 7 days of cell culture, there was a significant increase in cell viability in the control group compared to the BS, BS/SPG, BG and BG/SPG groups at a concentration of 100%, none of the materials showed signs of Genotoxicity and the materials were marked by alizarin red. For histological analysis, in vivo, 15 days after surgery, the groups presented granulation tissue and some areas of newly formed bone tissue. Thirty days after surgery, the groups revealed more mature newly formed bone tissue and progressive degradation of the material. All treated groups showed deposition of collagen organized in a network after 30 days. Immunohistochemical analysis demonstrated greater immunostaining for Runx-2 in all groups that received biomaterial implants, 15 and 30 days after surgery. For OPG analysis, an increase in immunostaining was observed for BS/SPG and BG/SPG compared to CG and BS and BG, 30 days after surgery. Therefore, it can be concluded that in vitro and in vivo studies indicated that BS and BS/SPG samples promoted greater cell viability and better biological properties in bone defects in the tibia of osteoporotic rats, highlighting the potential of composites associated with SPG to be used as bone grafts in regenerative medicine applications.
During the bone repair process, approximately 5 to 20% of cases may culminate in an abnormal consolidation process and result in a delay in bone union or non-union. Once this situation is established, it can lead to losses in the quality of life of these individuals, high rates of morbidity and mortality and high costs to the health system. Therefore, it is necessary to investigate treatments that have osteogenic potential and that have the ability to accelerate the bone repair process. Among these resources, the application of biomaterials can be highlighted, for example, bioactive materials, such as bioglass (BG) and those from marine sponges such as biosilica (BS) and spongin (SPG). Thus, the objective of the present study was to evaluate the biocompatibility of BS and BG composites associated with SPG, through in vitro and in vivo studies, using a bone defect model in the tibias of osteoporotic rats. For the in vitro study, MC3T3-E1 cells. L929 and CHOK-1 were analyzed using cell viability, comet assay, micronucleus test and alizarin red. In the in vivo study, the following analyzes were carried out: Histopathological, histomorphometric, collagen analysis and immunohistochemistry. The in vitro study demonstrated that, after 7 days of cell culture, there was a significant increase in cell viability in the control group compared to the BS, BS/SPG, BG and BG/SPG groups at a concentration of 100%, none of the materials showed signs of Genotoxicity and the materials were marked by alizarin red. For histological analysis, in vivo, 15 days after surgery, the groups presented granulation tissue and some areas of newly formed bone tissue. Thirty days after surgery, the groups revealed more mature newly formed bone tissue and progressive degradation of the material. All treated groups showed deposition of collagen organized in a network after 30 days. Immunohistochemical analysis demonstrated greater immunostaining for Runx-2 in all groups that received biomaterial implants, 15 and 30 days after surgery. For OPG analysis, an increase in immunostaining was observed for BS/SPG and BG/SPG compared to CG and BS and BG, 30 days after surgery. Therefore, it can be concluded that in vitro and in vivo studies indicated that BS and BS/SPG samples promoted greater cell viability and better biological properties in bone defects in the tibia of osteoporotic rats, highlighting the potential of composites associated with SPG to be used as bone grafts in regenerative medicine applications.
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Citação
CRUZ, Matheus de Almeida. Efeitos de scaffolds desenvolvidos a partir de compósitos bioativos extraídos de esponjas marinhas no processo de reparo ósseo de ratas osteoporóticas. 2023. 167 f. Tese (Doutorado em Bioprodutos e Bioprocessos) - Universidade Federal de São Paulo, Instituto de Saúde e Sociedade, Santos, 2023.