Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônico

Detalhes bibliográficos
Autor(a) principal: Carneiro, Bárbara Gressy Duarte Souza
Data de Publicação: 2020
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
Texto Completo: http://www.repositorio.ufc.br/handle/riufc/50323
Resumo: Bone is a dynamic tissue that presents important self-regenerative capacity, mediated by viable bone cells, adequate vascularity and growth factors. Nevertheless, in cases of congenital malformation, defects due trauma or tumor resection, and extensive craniofacial defects, where this bone capacity it is exceeded, biomaterial are often needed in order to support complete bone repair. Among the most used biomaterials in the clinical practice, Bio-Oss has stood out as a gold-standard bone substitute, despite having only an osteoconductor characteristics. Therefore, in situations where bone metabolism is compromised, as in patients under bisphosphonate therapy, becomes interesting the combination of xenogenous bone graft with biological amplifiers, such as growth factors. In this context, highlights platelet-rich fibrin (PRF), an immunological complex rich in platelets, leukocytes and growth factors embedded in a fibrin scaffold, that in spite of being well-known osteoinductor, still needs studies about its clinical efficacy in tissue regeneration, which led us to investigate the effect of the association between PRF and Bio-Oss on bone repair during bisphosphonate therapy. So, the aim of this work was to evaluate the effect of the association between PRF and Bio-Oss (BO) on bone remodeling in critical size-defect in rat calvaria treated with zoledronic acid (ZA). For this 36 Wistar male rats (Rattus norvegicus), weighing 300 g, were used. Twenty-four rats received previously ZA in single dose of 120 μg/kg, via s.c., similar to an osteoporosis treatment in humans. Seven days later, they were subjected to a surgery to create an 8 mm critical size defect in rat calvaria, and then, were divided into the following groups (n=6 animals/group) according to the treatment: ZA – did not receive bone graft; BO – had the defect filled with 0,04 ml of Bio-Oss; PRF - had the defect filled with 0,04 ml of PRF; BO+PRF - had the defect filled with BO and PRF. The animals of control group (C) (n=6) received saline solution and did not have the defect filled with any graft. All defects were recovered with a collagen membrane before suture. In addition, it was used 6 animals for blood donation in order to obtain the allogenic PRF. After 12 weeks, all animals were euthanized and calvaria was removed and submitted to histological and histomorphometric analyses. Data obtained were analyzed by GraphPad Prisma 8.3 software and were presented as mean±S.E.M using ANOVA followed by Tukey. The animals that received only ZA did not keep the calvaria anatomy, moreover they presented reduction of 34% on osteoblast, 79% on osteoclast, 77% on blood vessel counts and 35% reduction of total collagen, marked by reduction on type I collagen when compared to control (p<0,05). The treatment with Bio-Oss despite keeping the dimensional structure of the tissue, was not able to reverse the deleterious effect of ZA over bone cells and vessels. Meanwhile, the treatment with PRF caused increase of 37% on osteoblast, 72% on osteoclast, 74% on blood vessels counts and 39% of total collagen marked by increase of type I collagen (p<0,05). The association of BO+PRF still, showed better repair aspect, maintaining the tissue architecture. In this group (BO+PRF), BO particles were in smaller size with greater bone formation between these particles, or with fissure particles filled with bone tissue. In this way, we can conclude that the use of PRF associated to Bio-Oss can support the repair process by stimulation of bone remodeling, promoting angiogenesis, stimulating collagen production, keeping tissue anatomy and mitigating the negative effects on bone caused by zoledronic acid, suggesting that this association can be an important tool to the treatment of critical bone defects in patients using bisphosphonates.
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spelling Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônicoFibrina Rica em PlaquetasCrânioOsso e OssosBone is a dynamic tissue that presents important self-regenerative capacity, mediated by viable bone cells, adequate vascularity and growth factors. Nevertheless, in cases of congenital malformation, defects due trauma or tumor resection, and extensive craniofacial defects, where this bone capacity it is exceeded, biomaterial are often needed in order to support complete bone repair. Among the most used biomaterials in the clinical practice, Bio-Oss has stood out as a gold-standard bone substitute, despite having only an osteoconductor characteristics. Therefore, in situations where bone metabolism is compromised, as in patients under bisphosphonate therapy, becomes interesting the combination of xenogenous bone graft with biological amplifiers, such as growth factors. In this context, highlights platelet-rich fibrin (PRF), an immunological complex rich in platelets, leukocytes and growth factors embedded in a fibrin scaffold, that in spite of being well-known osteoinductor, still needs studies about its clinical efficacy in tissue regeneration, which led us to investigate the effect of the association between PRF and Bio-Oss on bone repair during bisphosphonate therapy. So, the aim of this work was to evaluate the effect of the association between PRF and Bio-Oss (BO) on bone remodeling in critical size-defect in rat calvaria treated with zoledronic acid (ZA). For this 36 Wistar male rats (Rattus norvegicus), weighing 300 g, were used. Twenty-four rats received previously ZA in single dose of 120 μg/kg, via s.c., similar to an osteoporosis treatment in humans. Seven days later, they were subjected to a surgery to create an 8 mm critical size defect in rat calvaria, and then, were divided into the following groups (n=6 animals/group) according to the treatment: ZA – did not receive bone graft; BO – had the defect filled with 0,04 ml of Bio-Oss; PRF - had the defect filled with 0,04 ml of PRF; BO+PRF - had the defect filled with BO and PRF. The animals of control group (C) (n=6) received saline solution and did not have the defect filled with any graft. All defects were recovered with a collagen membrane before suture. In addition, it was used 6 animals for blood donation in order to obtain the allogenic PRF. After 12 weeks, all animals were euthanized and calvaria was removed and submitted to histological and histomorphometric analyses. Data obtained were analyzed by GraphPad Prisma 8.3 software and were presented as mean±S.E.M using ANOVA followed by Tukey. The animals that received only ZA did not keep the calvaria anatomy, moreover they presented reduction of 34% on osteoblast, 79% on osteoclast, 77% on blood vessel counts and 35% reduction of total collagen, marked by reduction on type I collagen when compared to control (p<0,05). The treatment with Bio-Oss despite keeping the dimensional structure of the tissue, was not able to reverse the deleterious effect of ZA over bone cells and vessels. Meanwhile, the treatment with PRF caused increase of 37% on osteoblast, 72% on osteoclast, 74% on blood vessels counts and 39% of total collagen marked by increase of type I collagen (p<0,05). The association of BO+PRF still, showed better repair aspect, maintaining the tissue architecture. In this group (BO+PRF), BO particles were in smaller size with greater bone formation between these particles, or with fissure particles filled with bone tissue. In this way, we can conclude that the use of PRF associated to Bio-Oss can support the repair process by stimulation of bone remodeling, promoting angiogenesis, stimulating collagen production, keeping tissue anatomy and mitigating the negative effects on bone caused by zoledronic acid, suggesting that this association can be an important tool to the treatment of critical bone defects in patients using bisphosphonates.O osso é um órgão dinâmico que apresenta uma capacidade auto-regenerativa importante, mediada por células ósseas viáveis, vascularização adequada e fatores de crescimento. No entanto, em casos de má-formação congênita, defeitos devido a trauma ou ressecção de tumores e defeitos craniofaciais extensos, onde essa capacidade óssea é excedida, geralmente faz-se necessário o uso de biomateriais para auxiliar o completo reparo ósseo. Dentre os biomateriais utilizados na prática clínica, o Bio-Oss se destaca como um substituto ósseo padrão-ouro, apesar de apresentar apenas propriedades osteocondutoras. Por isso, em situações onde o metabolismo ósseo está comprometido, como no caso de pacientes usuários de bisfosfonatos, torna-se interessante a combinação deste enxerto xenógeno com amplificadores biológicos, por exemplo fatores de crescimento. Neste contexto, se destaca a fibrina rica em plaquetas (PRF), um complexo imunológico rico em plaquetas, leucócitos e fatores de crescimento embebidos em arcabouço de fibrina, que apesar de ser conhecidamente osteoindutora, ainda carece de estudos sobre sua eficácia clínica na regeneração tecidual o que nos leva a investigar o efeito da associação entre PRF e Bio-Oss no reparo ósseo durante terapia com bisfosfonatos. Assim, o objetivo desse estudo foi avaliar o efeito da associação de PRF e Bio-Oss (BO) na remodelação óssea de defeitos crítico em calvárias de ratos tratados ácido zoledrônico (AZ). Para isto, 36 ratos Wistar (Rattus norvegicus) machos, pesando 300 g, foram utilizados. Vinte e quatro ratos receberam previamente AZ em dose única de 120 μg/kg, via s.c., equivalente ao tratamento de osteoporose humano. Sete dias depois foram submetidos a cirurgia para criação de um defeito crítico de 8 mm na calvária e foram divididos nos seguintes grupos (n=6) de acordo com o tratamento: AZ – não recebeu enxerto; BO - teve o defeito preenchido com 0,04 ml de Bio-Oss; PRF - teve o defeito preenchido com 0,04 ml de PRF; BO+PRF - teve o defeito preenchido com BO e PRF. Os animais do grupo controle (C) (n=6) receberam solução salina e sem qualquer preenchimento do defeito. Todos os defeitos ósseos foram recobertos com membrana de colágeno bovino reabsorvível antes da sutura. Adicionalmente foi utilizado 6 animais doadores de sangue para obtenção do PRF alogênico. Após 12 semanas todos os animais foram eutanasiados e as calvárias foram removidas e submetidas às análises histológica e histomorfométrica. Os dados obtidos foram analisados através do software GraphPad Prisma 8.3, e foram apresentados com média±E.P.M utilizando os testes ANOVA seguido de Tukey. Os animais que receberam apenas AZ não mantiveram a anatomia da calota, além de apresentarem redução de 34% de osteoblastos, 79% de osteoclastos, 77% vasos sanguíneos e 35% colágeno total, marcada pela redução de colágeno tipo I quando comparado ao controle (p<0,05). O tratamento com Bio-Oss apesar de manter a estrutura dimensional do tecido não foi capaz de reverter os efeitos deletérios do AZ sobre células ósseas e vasos. No entanto, o tratamento com PRF promoveu aumento de 37% de osteoblastos, 72% de osteoclastos, 74% vasos sanguíneos e 39% colágeno total, marcado pelo aumento de colágeno tipo I (p<0,05). A associação BO+PRF ainda mostrou melhor aspecto de reparo pois além da manutenção da altura de tecido, as partículas de BO estavam em menor tamanho com maior formação óssea entre essas partículas, ou com invaginação de tecido ósseo em fissuras formadas nas próprias partículas de Bio-Oss. Desta forma podemos concluir que o uso de PRF associado ao Bio-Oss pode auxiliar o processo de reparo ósseo por estimular a remodelação óssea, promover angiogênese, estimular a produção de colágeno e manter a anatomia do tecido, atenuando os efeitos negativos em tecido ósseo causados pelo ácido zoledrônico, sugerindo que esta associação pode ser uma ferramenta importante no tratamento de defeitos ósseos críticos em pacientes que utilizam bisfosfonatos.Dutra, Paula Goes PinheiroGondim, Delane VianaCarneiro, Bárbara Gressy Duarte Souza2020-02-27T12:22:47Z2020-02-27T12:22:47Z2020-01-31info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfCARNEIRO, B. G. D. S. Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônico. 2020. 49 f. Dissertação (Mestrado em Odontologia) - Faculdade de Farmácia, Odontologia e Enfermagem, Universidade Federal do Ceará, 2020.http://www.repositorio.ufc.br/handle/riufc/50323porreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2020-02-27T12:22:47Zoai:repositorio.ufc.br:riufc/50323Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:55:48.385522Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.none.fl_str_mv Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônico
title Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônico
spellingShingle Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônico
Carneiro, Bárbara Gressy Duarte Souza
Fibrina Rica em Plaquetas
Crânio
Osso e Ossos
title_short Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônico
title_full Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônico
title_fullStr Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônico
title_full_unstemmed Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônico
title_sort Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônico
author Carneiro, Bárbara Gressy Duarte Souza
author_facet Carneiro, Bárbara Gressy Duarte Souza
author_role author
dc.contributor.none.fl_str_mv Dutra, Paula Goes Pinheiro
Gondim, Delane Viana
dc.contributor.author.fl_str_mv Carneiro, Bárbara Gressy Duarte Souza
dc.subject.por.fl_str_mv Fibrina Rica em Plaquetas
Crânio
Osso e Ossos
topic Fibrina Rica em Plaquetas
Crânio
Osso e Ossos
description Bone is a dynamic tissue that presents important self-regenerative capacity, mediated by viable bone cells, adequate vascularity and growth factors. Nevertheless, in cases of congenital malformation, defects due trauma or tumor resection, and extensive craniofacial defects, where this bone capacity it is exceeded, biomaterial are often needed in order to support complete bone repair. Among the most used biomaterials in the clinical practice, Bio-Oss has stood out as a gold-standard bone substitute, despite having only an osteoconductor characteristics. Therefore, in situations where bone metabolism is compromised, as in patients under bisphosphonate therapy, becomes interesting the combination of xenogenous bone graft with biological amplifiers, such as growth factors. In this context, highlights platelet-rich fibrin (PRF), an immunological complex rich in platelets, leukocytes and growth factors embedded in a fibrin scaffold, that in spite of being well-known osteoinductor, still needs studies about its clinical efficacy in tissue regeneration, which led us to investigate the effect of the association between PRF and Bio-Oss on bone repair during bisphosphonate therapy. So, the aim of this work was to evaluate the effect of the association between PRF and Bio-Oss (BO) on bone remodeling in critical size-defect in rat calvaria treated with zoledronic acid (ZA). For this 36 Wistar male rats (Rattus norvegicus), weighing 300 g, were used. Twenty-four rats received previously ZA in single dose of 120 μg/kg, via s.c., similar to an osteoporosis treatment in humans. Seven days later, they were subjected to a surgery to create an 8 mm critical size defect in rat calvaria, and then, were divided into the following groups (n=6 animals/group) according to the treatment: ZA – did not receive bone graft; BO – had the defect filled with 0,04 ml of Bio-Oss; PRF - had the defect filled with 0,04 ml of PRF; BO+PRF - had the defect filled with BO and PRF. The animals of control group (C) (n=6) received saline solution and did not have the defect filled with any graft. All defects were recovered with a collagen membrane before suture. In addition, it was used 6 animals for blood donation in order to obtain the allogenic PRF. After 12 weeks, all animals were euthanized and calvaria was removed and submitted to histological and histomorphometric analyses. Data obtained were analyzed by GraphPad Prisma 8.3 software and were presented as mean±S.E.M using ANOVA followed by Tukey. The animals that received only ZA did not keep the calvaria anatomy, moreover they presented reduction of 34% on osteoblast, 79% on osteoclast, 77% on blood vessel counts and 35% reduction of total collagen, marked by reduction on type I collagen when compared to control (p<0,05). The treatment with Bio-Oss despite keeping the dimensional structure of the tissue, was not able to reverse the deleterious effect of ZA over bone cells and vessels. Meanwhile, the treatment with PRF caused increase of 37% on osteoblast, 72% on osteoclast, 74% on blood vessels counts and 39% of total collagen marked by increase of type I collagen (p<0,05). The association of BO+PRF still, showed better repair aspect, maintaining the tissue architecture. In this group (BO+PRF), BO particles were in smaller size with greater bone formation between these particles, or with fissure particles filled with bone tissue. In this way, we can conclude that the use of PRF associated to Bio-Oss can support the repair process by stimulation of bone remodeling, promoting angiogenesis, stimulating collagen production, keeping tissue anatomy and mitigating the negative effects on bone caused by zoledronic acid, suggesting that this association can be an important tool to the treatment of critical bone defects in patients using bisphosphonates.
publishDate 2020
dc.date.none.fl_str_mv 2020-02-27T12:22:47Z
2020-02-27T12:22:47Z
2020-01-31
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.uri.fl_str_mv CARNEIRO, B. G. D. S. Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônico. 2020. 49 f. Dissertação (Mestrado em Odontologia) - Faculdade de Farmácia, Odontologia e Enfermagem, Universidade Federal do Ceará, 2020.
http://www.repositorio.ufc.br/handle/riufc/50323
identifier_str_mv CARNEIRO, B. G. D. S. Avaliação da associação de fibrina rica em plaquetas e enxerto xenógeno na reparo ósseo de defeitos criticos em calvárias de ratos tratados com ácido zoledrônico. 2020. 49 f. Dissertação (Mestrado em Odontologia) - Faculdade de Farmácia, Odontologia e Enfermagem, Universidade Federal do Ceará, 2020.
url http://www.repositorio.ufc.br/handle/riufc/50323
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