Enxerto ou implante homólogo na correção de defeito ósseo segmentar femoral em cães associado a inoculação da fração de células mononucleares da medula óssea

Detalhes bibliográficos
Autor(a) principal: Salbego, Fabiano Zanini
Data de Publicação: 2010
Tipo de documento: Tese
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
dARK ID: ark:/26339/0013000017mzd
Texto Completo: http://repositorio.ufsm.br/handle/1/4041
Resumo: Bone defects due to trauma, postoperative and mutilating surgery complications, associated to removal of neoplasia, are part of the routine of hospitals and veterinary clinics. Bone grafts, within all their variants, represent the main viable alternative in the structural correction of great bone losses. Currently, the search for alternatives to correct these problems and the great potential of cellular therapy, have led numerous researchers to ingress into this line of study. The present work aimed to evaluate the influence of the hydration of bone implants conserved in glycerin 98% over their biomechanical resistance; the viability of a bone marrow harvesting protocol, for application in the clinical surgical routine; and the effect of intralesional application of bone marrow mononuclear cell fraction, over the consolidation of graft-host interface of dogs submitted to segmental bone grafting, with bone implants conserved in glycerin 98%. This research was developed in two phases. The first, ex-vivo, in which 108 bone fragments conserved in glycerin 98% were separated in six different groups, according to hydration period, and later were submitted to axial load test for biomechanical resistance evaluation. The second phase, in-vivo, in which 20 mongrel dogs, with mean weight of 13kg, were divided in four different groups according to the type of graft employed and with or without application of bone marrow mononuclear cell fraction. All animals were submitted to a bone marrow harvesting protocol, obtaining a final volume of 5ml kg-1 of body weight. The collected bone marrow was analyzed by means of myelogram and later submitted to isolation of the mononuclear cell fraction by a technique already affirmed in the literature. A diaphyseal femoral bone defect was created and repaired with the same removed segment (groups I and III) or with a bone segment conserved in glycerin 98% (groups II and IV). The bone marrow mononuclear cell fraction, after isolation, was injected via intralesional route in dogs of the treated groups (I and II), whereas the control group (III and IV) received an injection of the same volume, however, of 0.9% sodium chloride solution. The bone consolidation progress of the graft-host interfaces was followed by serial radiographic exams up to 90 days after surgical intervention, whereas the presence of mononuclear cells was followed up to seven days after implantation, by detection of Qtracker-665 nanocrystal fluorescence, used as cell marker. According to the results obtained, it can be concluded that: hydration of bone implant conserved in glycerin 98% does not produce statistically significant alterations in the biomechanical resistance of cortical bone for the different evaluated times. However, the bone mineral density has a close relationship with this property; bone marrow harvesting protocol with reduced volume of 5ml kg-1 of body weight, obtained from different long bones, proved to be adequate for isolation of mononuclear cell fraction and for obtaining a cell button with adequate number of cells and high viability, essential for the success of its therapeutic application; the consolidation of proximal and distal graft-host interfaces at 90 days after surgery no showed clinical and radiographic difference in the groups control and treated with bone marrow mononuclear cell fraction. Nevertheless, it cannot be confirmed that the cellular therapy used in this study is not an effective bone healing adjuvant, as it is believed that the restriction of load deposition on the focus of the fracture, achieved by reduced postoperative exercise, has influenced directly on the result of the consolidation.
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spelling Enxerto ou implante homólogo na correção de defeito ósseo segmentar femoral em cães associado a inoculação da fração de células mononucleares da medula ósseaGraft or homologous implant in the correction of femoral segmental bone defect in dogs associated to inoculation of bone marrow mononuclear cell fractionCélulas mononuclearesMedula ósseaEnxerto ósseoFêmurCãoMononuclear cellsBone marrowBone graftFemurDogCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIABone defects due to trauma, postoperative and mutilating surgery complications, associated to removal of neoplasia, are part of the routine of hospitals and veterinary clinics. Bone grafts, within all their variants, represent the main viable alternative in the structural correction of great bone losses. Currently, the search for alternatives to correct these problems and the great potential of cellular therapy, have led numerous researchers to ingress into this line of study. The present work aimed to evaluate the influence of the hydration of bone implants conserved in glycerin 98% over their biomechanical resistance; the viability of a bone marrow harvesting protocol, for application in the clinical surgical routine; and the effect of intralesional application of bone marrow mononuclear cell fraction, over the consolidation of graft-host interface of dogs submitted to segmental bone grafting, with bone implants conserved in glycerin 98%. This research was developed in two phases. The first, ex-vivo, in which 108 bone fragments conserved in glycerin 98% were separated in six different groups, according to hydration period, and later were submitted to axial load test for biomechanical resistance evaluation. The second phase, in-vivo, in which 20 mongrel dogs, with mean weight of 13kg, were divided in four different groups according to the type of graft employed and with or without application of bone marrow mononuclear cell fraction. All animals were submitted to a bone marrow harvesting protocol, obtaining a final volume of 5ml kg-1 of body weight. The collected bone marrow was analyzed by means of myelogram and later submitted to isolation of the mononuclear cell fraction by a technique already affirmed in the literature. A diaphyseal femoral bone defect was created and repaired with the same removed segment (groups I and III) or with a bone segment conserved in glycerin 98% (groups II and IV). The bone marrow mononuclear cell fraction, after isolation, was injected via intralesional route in dogs of the treated groups (I and II), whereas the control group (III and IV) received an injection of the same volume, however, of 0.9% sodium chloride solution. The bone consolidation progress of the graft-host interfaces was followed by serial radiographic exams up to 90 days after surgical intervention, whereas the presence of mononuclear cells was followed up to seven days after implantation, by detection of Qtracker-665 nanocrystal fluorescence, used as cell marker. According to the results obtained, it can be concluded that: hydration of bone implant conserved in glycerin 98% does not produce statistically significant alterations in the biomechanical resistance of cortical bone for the different evaluated times. However, the bone mineral density has a close relationship with this property; bone marrow harvesting protocol with reduced volume of 5ml kg-1 of body weight, obtained from different long bones, proved to be adequate for isolation of mononuclear cell fraction and for obtaining a cell button with adequate number of cells and high viability, essential for the success of its therapeutic application; the consolidation of proximal and distal graft-host interfaces at 90 days after surgery no showed clinical and radiographic difference in the groups control and treated with bone marrow mononuclear cell fraction. Nevertheless, it cannot be confirmed that the cellular therapy used in this study is not an effective bone healing adjuvant, as it is believed that the restriction of load deposition on the focus of the fracture, achieved by reduced postoperative exercise, has influenced directly on the result of the consolidation.Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorAs falhas ósseas decorrentes de trauma, complicações pós-operatórias e cirurgias mutilantes, associadas à remoção de neoplasias, fazem parte da rotina dos hospitais e clínicas veterinárias. Os enxertos ósseos, dentre todas as suas variantes, representam a principal alternativa viável na correção estrutural de grandes perdas ósseas. Na atualidade, a busca por alternativas para corrigir estes problemas e o grande potencial da terapia celular, tem levado inúmeros pesquisadores a ingressar nesta linha de estudo. No presente trabalho, buscou-se avaliar a influência da hidratação de implantes ósseos conservados em glicerina 98% sobre a resistência biomecânica dos mesmos; a viabilidade de um protocolo de colheita de medula óssea, para aplicação na rotina clínico-cirúrgica; e o efeito da aplicação intralesional da fração de células mononucleares da medula óssea, sobre a consolidação da interface enxerto-hospedeiro de cães submetidos à enxertia óssea segmentar, com implantes ósseos conservados em glicerina 98%. Esta pesquisa foi desenvolvida em duas fases. A primeira, ex-vivo, onde 108 fragmentos ósseos conservados em glicerina 98% foram separados em seis diferentes grupos, de acordo com o período de hidratação, sendo posteriormente submetidos a teste de compressão axial para avaliação de sua resistência biomecânica. A segunda fase, in-vivo, onde 20 cães sem raça definida e peso médio de 13kg, foram separados em quatro diferentes grupos de acordo com o tipo de enxerto empregado e com a aplicação ou não da fração de células mononucleares da medula óssea. Todos os animais foram submetidos a um protocolo de colheita de medula óssea, obtendo-se um volume final de 5ml kg-1 de peso corporal. A medula óssea colhida foi analisada por meio de mielograma e posteriormente submetida ao isolamento da fração de células mononucleares por técnica já consagrada na literatura. Um defeito ósseo femoral diafisário foi criado e reparado com o próprio segmento removido (grupos I e III) ou com um segmento ósseo conservado em glicerina 98% (grupos II e IV). A fração de células mononucleares da medula óssea, após isolamento, foi injetada por via intralesional nos cães dos grupos tratados (I e II), enquanto os grupos controle (III e IV) receberam a injeção do mesmo volume, porém, de solução de cloreto de sódio a 0,9%. A progressão da consolidação óssea das interfaces enxerto-hospedeiro foi acompanhada por exames radiográficos seriados até os 90 dias após a intervenção cirúrgica, enquanto a presença das células mononucleares, até os primeiros sete dias após a implantação, foi acompanhada pela detecção da fluorescência do nanocristal Qtracker-665, utilizado como marcador celular. De acordo com os resultado obtidos, pode-se concluir que: a hidratação do implante ósseo conservado em glicerina 98%, não produz alteração estatisticamente significativa na resistência biomecânica do osso cortical para os diferentes tempos avaliados. Contudo, a densidade mineral óssea possui estreita relação com esta propriedade; O protocolo de colheita de medula óssea em volume reduzido a 5ml kg-1 de peso corporal, obtidos de diferentes ossos longos, demonstrou-se adequado para isolamento da fração de células mononucleares e obtenção de um botão celular com adequado número de células e alta viabilidade, essenciais ao sucesso de sua aplicação terapêutica; a consolidação das interfaces enxerto-hospedeiro proximal e distal aos 90 dias após a cirurgia não apresentou diferença clínica e radiográfica entre os grupos controle e tratados com a fração de células mononucleares da medula óssea. Porém, não se pode afirmar que a terapia celular empregada neste estudo, não seja bom adjuvante da cicatrização óssea, pois se acredita, que a restrição na deposição de carga no foco de fratura, proveniente do reduzido exercício pós-operatório, tenha influenciado diretamente no resultado da consolidação.Universidade Federal de Santa MariaBRMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaRaiser, Alceu Gasparhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727899U2Alievi, Marcelo Mellerhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4767491D6Contesini, Emerson Antoniohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4784012D5Pippi, Ney Luishttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783382P7Faria, Renato Xavierhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763636Z7Salbego, Fabiano Zanini2017-06-022017-06-022010-01-12info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfapplication/pdfSALBEGO, Fabiano Zanini. Graft or homologous implant in the correction of femoral segmental bone defect in dogs associated to inoculation of bone marrow mononuclear cell fraction. 2010. 211 f. Tese (Doutorado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2010.http://repositorio.ufsm.br/handle/1/4041ark:/26339/0013000017mzdporinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2017-07-25T14:03:23Zoai:repositorio.ufsm.br:1/4041Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2017-07-25T14:03:23Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Enxerto ou implante homólogo na correção de defeito ósseo segmentar femoral em cães associado a inoculação da fração de células mononucleares da medula óssea
Graft or homologous implant in the correction of femoral segmental bone defect in dogs associated to inoculation of bone marrow mononuclear cell fraction
title Enxerto ou implante homólogo na correção de defeito ósseo segmentar femoral em cães associado a inoculação da fração de células mononucleares da medula óssea
spellingShingle Enxerto ou implante homólogo na correção de defeito ósseo segmentar femoral em cães associado a inoculação da fração de células mononucleares da medula óssea
Salbego, Fabiano Zanini
Células mononucleares
Medula óssea
Enxerto ósseo
Fêmur
Cão
Mononuclear cells
Bone marrow
Bone graft
Femur
Dog
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Enxerto ou implante homólogo na correção de defeito ósseo segmentar femoral em cães associado a inoculação da fração de células mononucleares da medula óssea
title_full Enxerto ou implante homólogo na correção de defeito ósseo segmentar femoral em cães associado a inoculação da fração de células mononucleares da medula óssea
title_fullStr Enxerto ou implante homólogo na correção de defeito ósseo segmentar femoral em cães associado a inoculação da fração de células mononucleares da medula óssea
title_full_unstemmed Enxerto ou implante homólogo na correção de defeito ósseo segmentar femoral em cães associado a inoculação da fração de células mononucleares da medula óssea
title_sort Enxerto ou implante homólogo na correção de defeito ósseo segmentar femoral em cães associado a inoculação da fração de células mononucleares da medula óssea
author Salbego, Fabiano Zanini
author_facet Salbego, Fabiano Zanini
author_role author
dc.contributor.none.fl_str_mv Raiser, Alceu Gaspar
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727899U2
Alievi, Marcelo Meller
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4767491D6
Contesini, Emerson Antonio
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4784012D5
Pippi, Ney Luis
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783382P7
Faria, Renato Xavier
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763636Z7
dc.contributor.author.fl_str_mv Salbego, Fabiano Zanini
dc.subject.por.fl_str_mv Células mononucleares
Medula óssea
Enxerto ósseo
Fêmur
Cão
Mononuclear cells
Bone marrow
Bone graft
Femur
Dog
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
topic Células mononucleares
Medula óssea
Enxerto ósseo
Fêmur
Cão
Mononuclear cells
Bone marrow
Bone graft
Femur
Dog
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Bone defects due to trauma, postoperative and mutilating surgery complications, associated to removal of neoplasia, are part of the routine of hospitals and veterinary clinics. Bone grafts, within all their variants, represent the main viable alternative in the structural correction of great bone losses. Currently, the search for alternatives to correct these problems and the great potential of cellular therapy, have led numerous researchers to ingress into this line of study. The present work aimed to evaluate the influence of the hydration of bone implants conserved in glycerin 98% over their biomechanical resistance; the viability of a bone marrow harvesting protocol, for application in the clinical surgical routine; and the effect of intralesional application of bone marrow mononuclear cell fraction, over the consolidation of graft-host interface of dogs submitted to segmental bone grafting, with bone implants conserved in glycerin 98%. This research was developed in two phases. The first, ex-vivo, in which 108 bone fragments conserved in glycerin 98% were separated in six different groups, according to hydration period, and later were submitted to axial load test for biomechanical resistance evaluation. The second phase, in-vivo, in which 20 mongrel dogs, with mean weight of 13kg, were divided in four different groups according to the type of graft employed and with or without application of bone marrow mononuclear cell fraction. All animals were submitted to a bone marrow harvesting protocol, obtaining a final volume of 5ml kg-1 of body weight. The collected bone marrow was analyzed by means of myelogram and later submitted to isolation of the mononuclear cell fraction by a technique already affirmed in the literature. A diaphyseal femoral bone defect was created and repaired with the same removed segment (groups I and III) or with a bone segment conserved in glycerin 98% (groups II and IV). The bone marrow mononuclear cell fraction, after isolation, was injected via intralesional route in dogs of the treated groups (I and II), whereas the control group (III and IV) received an injection of the same volume, however, of 0.9% sodium chloride solution. The bone consolidation progress of the graft-host interfaces was followed by serial radiographic exams up to 90 days after surgical intervention, whereas the presence of mononuclear cells was followed up to seven days after implantation, by detection of Qtracker-665 nanocrystal fluorescence, used as cell marker. According to the results obtained, it can be concluded that: hydration of bone implant conserved in glycerin 98% does not produce statistically significant alterations in the biomechanical resistance of cortical bone for the different evaluated times. However, the bone mineral density has a close relationship with this property; bone marrow harvesting protocol with reduced volume of 5ml kg-1 of body weight, obtained from different long bones, proved to be adequate for isolation of mononuclear cell fraction and for obtaining a cell button with adequate number of cells and high viability, essential for the success of its therapeutic application; the consolidation of proximal and distal graft-host interfaces at 90 days after surgery no showed clinical and radiographic difference in the groups control and treated with bone marrow mononuclear cell fraction. Nevertheless, it cannot be confirmed that the cellular therapy used in this study is not an effective bone healing adjuvant, as it is believed that the restriction of load deposition on the focus of the fracture, achieved by reduced postoperative exercise, has influenced directly on the result of the consolidation.
publishDate 2010
dc.date.none.fl_str_mv 2010-01-12
2017-06-02
2017-06-02
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv SALBEGO, Fabiano Zanini. Graft or homologous implant in the correction of femoral segmental bone defect in dogs associated to inoculation of bone marrow mononuclear cell fraction. 2010. 211 f. Tese (Doutorado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2010.
http://repositorio.ufsm.br/handle/1/4041
dc.identifier.dark.fl_str_mv ark:/26339/0013000017mzd
identifier_str_mv SALBEGO, Fabiano Zanini. Graft or homologous implant in the correction of femoral segmental bone defect in dogs associated to inoculation of bone marrow mononuclear cell fraction. 2010. 211 f. Tese (Doutorado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2010.
ark:/26339/0013000017mzd
url http://repositorio.ufsm.br/handle/1/4041
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
BR
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
publisher.none.fl_str_mv Universidade Federal de Santa Maria
BR
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
dc.source.none.fl_str_mv reponame:Manancial - Repositório Digital da UFSM
instname:Universidade Federal de Santa Maria (UFSM)
instacron:UFSM
instname_str Universidade Federal de Santa Maria (UFSM)
instacron_str UFSM
institution UFSM
reponame_str Manancial - Repositório Digital da UFSM
collection Manancial - Repositório Digital da UFSM
repository.name.fl_str_mv Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)
repository.mail.fl_str_mv atendimento.sib@ufsm.br||tedebc@gmail.com
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