Biocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivo

Detalhes bibliográficos
Autor(a) principal: Kido, Hueliton Wilian
Data de Publicação: 2011
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFSCAR
Texto Completo: https://repositorio.ufscar.br/handle/ufscar/6995
Resumo: Due to limited availability of autogenous bone and of the risks associated with the use of bone allografts, new synthetic materials have been developed in order to replace the bone tissue lost due to trauma or pathological process. The bioactive materials in the form of scaffolds are synthetic materials promising for bone grafting. Several studies suggest that these biomaterials are able to stimulate the proliferation of osteoblasts and osteogenesis at the site of fracture. However, the feasibility of these biomaterials to a clinical application requires the investigation of their biocompatibility. In this context, this study aimed to evaluate the biocompatibility of a scaffold synthesized from a fully crystallized glass-ceramic bioactive quaternary system P2O5-Na2O-CaO-SiO2 (Biosilicate®), through histopathological analysis of the biomaterial implanted in the subcutaneous tissue of rats and the cytotoxicity and genotoxicity analysis of the biomaterial in cell cultures (OSTEO-1 and L929 cells). Histopathologic analysis of the biomaterial was performed using 65 Wistar rats male (210- 260 g), randomly divided into two groups, Control group (n = 3 animals per period) and Biosilicate group (n = 10 animals per period), evaluated at 7, 15, 30, 45 and 60 days after surgery. The animals of Biosilicate group underwent surgery and received a subcutaneous implant of Biosilicate® scaffolds. The animals of Control group underwent surgery but did not receive any biomaterial implant. The cytotoxicity analysis was performed to assess the effect of products leaching from Biosilicate® scaffolds (extracts) on cellular proliferation (MTT). The extracts were evaluated in various concentrations (100, 50, 25 and 12.5%) in experimental periods of 24, 72 and 120 hours in two cell lines (OSTEO-1 and L929). The genotoxicity analysis (comet assay) was performed to assess DNA damage in cells OSTEO-1 and L929 grown in contact with the Biosilicate® scaffolds in different periods of 24, 72 and 96 horas. The statistical analysis of parametrics data was performed by analysis of variance (ANOVA) followed by Tukey post-hoc and the analysis of nonparametrics data was performed by Mann-Whitney test. Both statistical tests were performed with a significance level of 5%. The results of histopathological analysis showed that the animals of the Control group did not present inflammation process, necrotic tissue and fibrous tissue. The animals of Biosilicato group showed a granulation tissue after 7 days of implantation. In the other periods (15, 30, 45 and 60 days) a chronic inflammation process of foreign body, marked by the presence of fibrous tissue and giant cells was observed. No infection or necrotic tissue was observed in any animal. In the analysis of cytotoxicity, it was observed that extracts of Biosilicato® scaffolds did not have any significant effect in reducing cell proliferation OSTEO-1 and L929, and that lower concentrations of the extracts (12.5 and 25%) stimulated the proliferation of both cells in periods of 72 and 120 hours. The analysis of genotoxicity showed that the Biosilicate® scaffolds did not induce DNA damage in the cell lines tested in all experimental periods. The results of this study showed that the Biosilicate® scaffolds presented biocompatibility in vivo and in vitro.
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spelling Kido, Hueliton WilianPeitl Filho, Oscarhttp://lattes.cnpq.br/3007441705652783http://lattes.cnpq.br/29135519444093231d4b7715-ce19-4b39-880b-7ab09a22472b2016-08-17T18:39:42Z2012-04-032016-08-17T18:39:42Z2011-08-29KIDO, Hueliton Wilian. Biocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivo. 2011. 75 f. Dissertação (Mestrado em Multidisciplinar) - Universidade Federal de São Carlos, São Carlos, 2011.https://repositorio.ufscar.br/handle/ufscar/6995Due to limited availability of autogenous bone and of the risks associated with the use of bone allografts, new synthetic materials have been developed in order to replace the bone tissue lost due to trauma or pathological process. The bioactive materials in the form of scaffolds are synthetic materials promising for bone grafting. Several studies suggest that these biomaterials are able to stimulate the proliferation of osteoblasts and osteogenesis at the site of fracture. However, the feasibility of these biomaterials to a clinical application requires the investigation of their biocompatibility. In this context, this study aimed to evaluate the biocompatibility of a scaffold synthesized from a fully crystallized glass-ceramic bioactive quaternary system P2O5-Na2O-CaO-SiO2 (Biosilicate®), through histopathological analysis of the biomaterial implanted in the subcutaneous tissue of rats and the cytotoxicity and genotoxicity analysis of the biomaterial in cell cultures (OSTEO-1 and L929 cells). Histopathologic analysis of the biomaterial was performed using 65 Wistar rats male (210- 260 g), randomly divided into two groups, Control group (n = 3 animals per period) and Biosilicate group (n = 10 animals per period), evaluated at 7, 15, 30, 45 and 60 days after surgery. The animals of Biosilicate group underwent surgery and received a subcutaneous implant of Biosilicate® scaffolds. The animals of Control group underwent surgery but did not receive any biomaterial implant. The cytotoxicity analysis was performed to assess the effect of products leaching from Biosilicate® scaffolds (extracts) on cellular proliferation (MTT). The extracts were evaluated in various concentrations (100, 50, 25 and 12.5%) in experimental periods of 24, 72 and 120 hours in two cell lines (OSTEO-1 and L929). The genotoxicity analysis (comet assay) was performed to assess DNA damage in cells OSTEO-1 and L929 grown in contact with the Biosilicate® scaffolds in different periods of 24, 72 and 96 horas. The statistical analysis of parametrics data was performed by analysis of variance (ANOVA) followed by Tukey post-hoc and the analysis of nonparametrics data was performed by Mann-Whitney test. Both statistical tests were performed with a significance level of 5%. The results of histopathological analysis showed that the animals of the Control group did not present inflammation process, necrotic tissue and fibrous tissue. The animals of Biosilicato group showed a granulation tissue after 7 days of implantation. In the other periods (15, 30, 45 and 60 days) a chronic inflammation process of foreign body, marked by the presence of fibrous tissue and giant cells was observed. No infection or necrotic tissue was observed in any animal. In the analysis of cytotoxicity, it was observed that extracts of Biosilicato® scaffolds did not have any significant effect in reducing cell proliferation OSTEO-1 and L929, and that lower concentrations of the extracts (12.5 and 25%) stimulated the proliferation of both cells in periods of 72 and 120 hours. The analysis of genotoxicity showed that the Biosilicate® scaffolds did not induce DNA damage in the cell lines tested in all experimental periods. The results of this study showed that the Biosilicate® scaffolds presented biocompatibility in vivo and in vitro.Devido a limitada disponibilidade de osso autógeno e dos riscos associados ao uso de osso alógeno, novos materiais sintéticos vêm sendo desenvolvidos com o objetivo de substituir o tecido ósseo perdido em decorrência de traumatismos ou processos patológicos. Os materiais bioativos na forma de scaffolds são materiais sintéticos promissores para enxertia óssea. Vários estudos sugerem que estes biomateriais são capazes de estimular a proliferação de osteoblastos e a osteogênese no local da fratura. No entanto, a viabilização destes biomateriais a uma aplicação clínica requer o emprego de testes que avaliem a sua biocompatibilidade. Dentro deste contexto, o presente estudo teve como objetivo avaliar a biocompatibilidade do scaffold sintetizado a partir de uma vitrocerâmica bioativa totalmente cristalizada do sistema quaternário P2O5-Na2O-CaO-SiO2 (Biosilicato®), por meio da análise histopatológica do biomaterial implantado no tecido subcutâneo de ratos, e pelas análises de citotoxicidade e genotoxicidade do biomaterial em cultura de células da linhagem OSTEO-1 e L929. A análise histopatológica do biomaterial foi realizada utilizando 65 ratos machos da linhagem Wistar (210-260 g), distribuídos aleatoriamente em dois grupos, Controle (n = 3 animais por período) e Biosilicato (n = 10 animais por período), avaliados em períodos distintos de 7, 15, 30, 45 e 60 dias. Os animais do grupo Biosilicato foram submetidos a uma cirurgia no tecido subcutâneo e receberam um implante de scaffold de Biosilicato®. Os animais do grupo Controle foram submetidos à mesma cirurgia, mas não receberam o implante do biomaterial. A análise de citotoxicidade foi realizada para avaliar os efeitos dos produtos da lixiviação dos scaffolds de Biosilicato® (extratos) na proliferação celular pelo ensaio MTT. Os extratos foram avaliados em várias concentrações (100, 50, 25 e 12,5%) em períodos experimentais de 24, 72 e 120 horas, utilizando duas linhagens celulares (OSTEO-1 e L929). A análise de genotoxicidade (ensaio cometa) foi realizada para avaliar os danos no DNA de células OSTEO-1 e L929 cultivadas em contato com scaffolds de Biosilicato® em períodos distintos de 24, 72 e 96 horas. A análise estatística dos dados paramétricos foi realizada pelo teste de variância (ANOVA), seguido do post-hoc de Tukey, e a análise dos dados não paramétricos foi realizada pelo teste de Mann-Whitney. Ambos os testes estatísticos foram realizados com nível de significância de 5%. Os resultados da análise histopatológica demonstraram que os animais do grupo Controle não apresentaram processo inflamatório, tecido necrótico ou tecido fibroso. Já os animais do grupo Biosilicato apresentaram um tecido de granulação após 7 dias de implantação e nos demais períodos (15, 30, 45 e 60 dias) apresentaram um processo inflamatório crônico de corpo estranho, marcado pela presença de tecido fibroso e células gigantes multinucleadas. Em todos os animais avaliados não foi evidenciado foco de infecção ou tecido necrótico. Na análise de citotoxicidade foi observado que os extratos dos scaffolds de Biosilicato® não possuem efeito significativo na redução da proliferação de células OSTEO-1 e L929, e que as menores concentrações dos extratos (12,5 e 25%) estimularam a proliferação de ambas às células nos períodos de 72 e 120 horas. Na análise de genotoxicidade foi evidenciado que os scaffolds de Biosilicato® não induzem danos do DNA de células de ambas às linhagens testadas em todos os períodos experimentais. Os resultados obtidos neste estudo demonstraram que os scaffolds de Biosilicato® apresentaram biocompatibilidade em experimentos in vivo e in vitro.Universidade Federal de Minas Geraisapplication/pdfporUniversidade Federal de São CarlosPrograma de Pós-Graduação em Biotecnologia - PPGBiotecUFSCarBRBiotecnologiaScaffoldBiomateriaisbiocompatibilidadereposta tecidualmaterial bioativobiocompatibilitytissue responsebioactive materialscaffoldbiomaterialCIENCIAS BIOLOGICAS::MORFOLOGIABiocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivoinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis-1-18c905e40-5692-43a9-926c-3ae9c7bdedfcinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINAL4169.pdfapplication/pdf1866763https://repositorio.ufscar.br/bitstream/ufscar/6995/1/4169.pdf38d468ca7232537d6952ee24e68d0593MD51TEXT4169.pdf.txt4169.pdf.txtExtracted texttext/plain106467https://repositorio.ufscar.br/bitstream/ufscar/6995/2/4169.pdf.txta682a2b74e2daf2cab0d18cb5e434530MD52THUMBNAIL4169.pdf.jpg4169.pdf.jpgIM Thumbnailimage/jpeg6533https://repositorio.ufscar.br/bitstream/ufscar/6995/3/4169.pdf.jpgd0bc5f0115f6980d58a7018535668f2cMD53ufscar/69952023-09-18 18:30:33.832oai:repositorio.ufscar.br:ufscar/6995Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:30:33Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false
dc.title.por.fl_str_mv Biocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivo
title Biocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivo
spellingShingle Biocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivo
Kido, Hueliton Wilian
Biotecnologia
Scaffold
Biomateriais
biocompatibilidade
reposta tecidual
material bioativo
biocompatibility
tissue response
bioactive material
scaffold
biomaterial
CIENCIAS BIOLOGICAS::MORFOLOGIA
title_short Biocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivo
title_full Biocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivo
title_fullStr Biocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivo
title_full_unstemmed Biocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivo
title_sort Biocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivo
author Kido, Hueliton Wilian
author_facet Kido, Hueliton Wilian
author_role author
dc.contributor.authorlattes.por.fl_str_mv http://lattes.cnpq.br/2913551944409323
dc.contributor.author.fl_str_mv Kido, Hueliton Wilian
dc.contributor.advisor1.fl_str_mv Peitl Filho, Oscar
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/3007441705652783
dc.contributor.authorID.fl_str_mv 1d4b7715-ce19-4b39-880b-7ab09a22472b
contributor_str_mv Peitl Filho, Oscar
dc.subject.por.fl_str_mv Biotecnologia
Scaffold
Biomateriais
biocompatibilidade
reposta tecidual
material bioativo
topic Biotecnologia
Scaffold
Biomateriais
biocompatibilidade
reposta tecidual
material bioativo
biocompatibility
tissue response
bioactive material
scaffold
biomaterial
CIENCIAS BIOLOGICAS::MORFOLOGIA
dc.subject.eng.fl_str_mv biocompatibility
tissue response
bioactive material
scaffold
biomaterial
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS::MORFOLOGIA
description Due to limited availability of autogenous bone and of the risks associated with the use of bone allografts, new synthetic materials have been developed in order to replace the bone tissue lost due to trauma or pathological process. The bioactive materials in the form of scaffolds are synthetic materials promising for bone grafting. Several studies suggest that these biomaterials are able to stimulate the proliferation of osteoblasts and osteogenesis at the site of fracture. However, the feasibility of these biomaterials to a clinical application requires the investigation of their biocompatibility. In this context, this study aimed to evaluate the biocompatibility of a scaffold synthesized from a fully crystallized glass-ceramic bioactive quaternary system P2O5-Na2O-CaO-SiO2 (Biosilicate®), through histopathological analysis of the biomaterial implanted in the subcutaneous tissue of rats and the cytotoxicity and genotoxicity analysis of the biomaterial in cell cultures (OSTEO-1 and L929 cells). Histopathologic analysis of the biomaterial was performed using 65 Wistar rats male (210- 260 g), randomly divided into two groups, Control group (n = 3 animals per period) and Biosilicate group (n = 10 animals per period), evaluated at 7, 15, 30, 45 and 60 days after surgery. The animals of Biosilicate group underwent surgery and received a subcutaneous implant of Biosilicate® scaffolds. The animals of Control group underwent surgery but did not receive any biomaterial implant. The cytotoxicity analysis was performed to assess the effect of products leaching from Biosilicate® scaffolds (extracts) on cellular proliferation (MTT). The extracts were evaluated in various concentrations (100, 50, 25 and 12.5%) in experimental periods of 24, 72 and 120 hours in two cell lines (OSTEO-1 and L929). The genotoxicity analysis (comet assay) was performed to assess DNA damage in cells OSTEO-1 and L929 grown in contact with the Biosilicate® scaffolds in different periods of 24, 72 and 96 horas. The statistical analysis of parametrics data was performed by analysis of variance (ANOVA) followed by Tukey post-hoc and the analysis of nonparametrics data was performed by Mann-Whitney test. Both statistical tests were performed with a significance level of 5%. The results of histopathological analysis showed that the animals of the Control group did not present inflammation process, necrotic tissue and fibrous tissue. The animals of Biosilicato group showed a granulation tissue after 7 days of implantation. In the other periods (15, 30, 45 and 60 days) a chronic inflammation process of foreign body, marked by the presence of fibrous tissue and giant cells was observed. No infection or necrotic tissue was observed in any animal. In the analysis of cytotoxicity, it was observed that extracts of Biosilicato® scaffolds did not have any significant effect in reducing cell proliferation OSTEO-1 and L929, and that lower concentrations of the extracts (12.5 and 25%) stimulated the proliferation of both cells in periods of 72 and 120 hours. The analysis of genotoxicity showed that the Biosilicate® scaffolds did not induce DNA damage in the cell lines tested in all experimental periods. The results of this study showed that the Biosilicate® scaffolds presented biocompatibility in vivo and in vitro.
publishDate 2011
dc.date.issued.fl_str_mv 2011-08-29
dc.date.available.fl_str_mv 2012-04-03
2016-08-17T18:39:42Z
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dc.identifier.citation.fl_str_mv KIDO, Hueliton Wilian. Biocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivo. 2011. 75 f. Dissertação (Mestrado em Multidisciplinar) - Universidade Federal de São Carlos, São Carlos, 2011.
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identifier_str_mv KIDO, Hueliton Wilian. Biocompatibilidade da vitrocêramica bioativa (Biosilicato®): análises in vitro e in vivo. 2011. 75 f. Dissertação (Mestrado em Multidisciplinar) - Universidade Federal de São Carlos, São Carlos, 2011.
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