Estudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratos
Autor(a) principal: | |
---|---|
Data de Publicação: | 2009 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/1310 |
Resumo: | INTRODUCTION: The incisional hernia (IH) is one of the most frequent complications after laparotomy, causing high rates of re-operations. Advances in suture material, in the guidelines for incisions, in the techniques of closure and use of dentures (screens), have failed in the elimination of this surgical complication. In this context, advances in the understanding of biological and structural changes of collagen may offer a new strategy in the treatment of IH, since the matrix disorders that induces the formation of primary hernia probably also affect the wound healing in laparotomies. OBJECTIVE: To investigate the effect of a recombinant desintegrin (DisBa-01) on the abdominal muscle healing process with incisional hernia (IH) regarding the fibroblasts and vessels proliferation and matrix metalopeptidase activity as well. The role of α2 and anti- αvβ3 integrins were investigated by antibody competition. METHODS: IH was induced by sectioning of the midline in 20 Wistar rats, divided into two groups: control that received the topical application of phosphate buffered saline (PBS) and DisBa-01 protein (0.5mg/Kg) dissolved in PBS. Another 10 rats were divided in two groups: one received anti-αVβ3 antibodies and the second anti-α2 subunit antibodies. At the end of 14 days, the animals were sacrificed and muscle fragments were stored in liquid nitrogen for determination of MMPs by zymography. The remaining fragments of the control and DisBa-01 groups were fixed in 10% formol in PBS (v/v). Blood vessels density, mononuclear cells, fibroblasts and collagen fibers were studied by optical microscopy in blue reticuline and Masson-trichrome. Data was analyzed by parametric tests were applied with critical level of 5%. RESULTS: All animals after surgical induction developed IH. However, after 14 days the persistence of the opened hernial ring in the controls but not in the DisBa-01 group. This protein induced the formation of a membrane that occluded the entire hernial ring. A significant increase in the number of mononuclear cells, fibroblasts, new vessels and MMP-2 activity was observed in DisBa-01 group (p<0.05). DisBa-01 group significant increased the collagen content. Anti-αVβ3 integrin antibodies produced similar results but not anti-α2 integrin blocking antibodies. CONCLUSION: These results indicate that DisBa-01 promoted vascular and fibroblastic cell proliferation and monocytic, and matrix remodeling. In addition, these results strongly indicate that αVβ3 integrin has an important role in the control of would healing and the blocking of this integrin may be an interesting therapeutical strategy in tissue repair conditions, including IH. |
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Oliveira, Claudio Ricardo deAraújo, Heloísa Sobreiro Selistre dehttp://genos.cnpq.br:12010/dwlattes/owa/prc_imp_cv_int?f_cod=K4787059Y2http://lattes.cnpq.br/50917057456137265f4304d8-aa9e-4739-9956-4db5e4d5b70c2016-06-02T19:22:51Z2009-08-272016-06-02T19:22:51Z2009-04-29OLIVEIRA, Claudio Ricardo de. Estudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratos. 2009. 92 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de São Carlos, São Carlos, 2009.https://repositorio.ufscar.br/handle/ufscar/1310INTRODUCTION: The incisional hernia (IH) is one of the most frequent complications after laparotomy, causing high rates of re-operations. Advances in suture material, in the guidelines for incisions, in the techniques of closure and use of dentures (screens), have failed in the elimination of this surgical complication. In this context, advances in the understanding of biological and structural changes of collagen may offer a new strategy in the treatment of IH, since the matrix disorders that induces the formation of primary hernia probably also affect the wound healing in laparotomies. OBJECTIVE: To investigate the effect of a recombinant desintegrin (DisBa-01) on the abdominal muscle healing process with incisional hernia (IH) regarding the fibroblasts and vessels proliferation and matrix metalopeptidase activity as well. The role of α2 and anti- αvβ3 integrins were investigated by antibody competition. METHODS: IH was induced by sectioning of the midline in 20 Wistar rats, divided into two groups: control that received the topical application of phosphate buffered saline (PBS) and DisBa-01 protein (0.5mg/Kg) dissolved in PBS. Another 10 rats were divided in two groups: one received anti-αVβ3 antibodies and the second anti-α2 subunit antibodies. At the end of 14 days, the animals were sacrificed and muscle fragments were stored in liquid nitrogen for determination of MMPs by zymography. The remaining fragments of the control and DisBa-01 groups were fixed in 10% formol in PBS (v/v). Blood vessels density, mononuclear cells, fibroblasts and collagen fibers were studied by optical microscopy in blue reticuline and Masson-trichrome. Data was analyzed by parametric tests were applied with critical level of 5%. RESULTS: All animals after surgical induction developed IH. However, after 14 days the persistence of the opened hernial ring in the controls but not in the DisBa-01 group. This protein induced the formation of a membrane that occluded the entire hernial ring. A significant increase in the number of mononuclear cells, fibroblasts, new vessels and MMP-2 activity was observed in DisBa-01 group (p<0.05). DisBa-01 group significant increased the collagen content. Anti-αVβ3 integrin antibodies produced similar results but not anti-α2 integrin blocking antibodies. CONCLUSION: These results indicate that DisBa-01 promoted vascular and fibroblastic cell proliferation and monocytic, and matrix remodeling. In addition, these results strongly indicate that αVβ3 integrin has an important role in the control of would healing and the blocking of this integrin may be an interesting therapeutical strategy in tissue repair conditions, including IH.INTRODUÇÃO: a hérnia incisional (HI) é uma das complicações mais freqüentes após laparotomia, causando altos índices de re-operações. Avanços nos materiais de suturas, nas orientações de incisões, nas técnicas de fechamento e do uso de próteses (telas), têm falhado para a eliminação desta complicação cirúrgica. Neste contexto, avanços no entendimento das alterações biológicas e estruturais do colágeno podem oferecer uma nova estratégia no tratamento da HI, uma vez que as desordens da matriz que induz a formação da hérnia primária provavelmente também prejudicam a cicatrização da ferida operatória nas laparotomias. OBJETIVO: Investigar os efeitos da desintegrina recombinante DisBa-01, sobre o processo de cicatrização da musculatura da parede abdominal com hérnia incisional (HI) com relação a proliferação de fibroblastos, vasos e a atividade das metalopeptidases de matriz. A função das integrinas anti-subunidade α2 e anti- αVβ3 foram investigadas pelo bloqueio de anticorpos. MÉTODOS: Foi induzida a HI por secção da linha alba em 20 ratos Wistar, distribuídos em dois grupos: controle que recebeu a aplicação tópica de solução salina tamponada com fosfato (PBS), e DisBa-01 (0,5mg/Kg) dissolvida em PBS. Outros 10 ratos foram divididos em dois grupos: um recebeu anti-αVβ3 e outro anti-subunidade α2. Ao final, de 14 dias, os animais foram sacrificados e fragmentos do músculo dos 04 grupos foram armazenados em nitrogênio liquido para determinação de MMPs por zimografia. O restante dos fragmentos dos grupos controle e DisBa-01, foram fixados em formol a 10% em PBS (v/v). A densidade de vasos sangüíneos, células mononucleares, fibroblastos e de fibras colágenas foram estudadas por microscopia óptica em reticulina e Tricromo de Masson. Para analise dos dados foram aplicados testes paramétricos com nível critico de 5%. RESULTADOS: Todos os animais após indução cirúrgica desenvolveram HI. Entretanto, após 14 dias nos animais do grupo Controle houve persistência do anel herniário aberto e nos animais do grupo DisBa-01 foi observado a formação de membrana que ocluía todo o anel herniário. Foi observado aumento significativo das células mononucleares, fibroblastos, neovascularização e da atividade da MMP-2 no grupo DisBa-01 (p<0,05). Houve aumento significativo da colagenização no grupo DisBa-01 em comparação ao controle. O grupo anti-αVβ3 produziu resultados similares a proteína DisBa-01 mas não no grupo bloqueado anti-subunidade α2. CONCLUSÃO: Estes resultados indicam que a DisBa-01 promoveu proliferação vascular, celular fibroblástica e monocítica, e remodelamento da MEC. Adicionalmente, estes resultados fortemente indicam que a integrina αVβ3 tem importante função no controle da ferida cirúrgica e o bloqueio desta integrina pode ser uma interessante estratégia terapêutica nas condições de reparo tecidual, incluindo a HI.application/pdfporUniversidade Federal de São CarlosPrograma Interinstitucional de Pós-Graduação em Ciências Fisiológicas - PIPGCFUFSCarBRHérnia ventralAbdomenDisBa-01Cicatrização de feridasRatosVentral hernialAbdomenDisBa-01Wound healingRatsCIENCIAS BIOLOGICAS::FISIOLOGIAEstudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratosinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis-1-1b61211db-d955-45b7-886e-a0cefb89980finfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINAL2443.pdfapplication/pdf1570096https://repositorio.ufscar.br/bitstream/ufscar/1310/1/2443.pdf27db268f8c997a54559bd609fc4ac9ceMD51THUMBNAIL2443.pdf.jpg2443.pdf.jpgIM Thumbnailimage/jpeg11360https://repositorio.ufscar.br/bitstream/ufscar/1310/2/2443.pdf.jpgeb4f72efc81fac0565e47105a4b1257eMD52ufscar/13102023-09-18 18:31:18.339oai:repositorio.ufscar.br:ufscar/1310Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:18Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Estudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratos |
title |
Estudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratos |
spellingShingle |
Estudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratos Oliveira, Claudio Ricardo de Hérnia ventral Abdomen DisBa-01 Cicatrização de feridas Ratos Ventral hernial Abdomen DisBa-01 Wound healing Rats CIENCIAS BIOLOGICAS::FISIOLOGIA |
title_short |
Estudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratos |
title_full |
Estudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratos |
title_fullStr |
Estudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratos |
title_full_unstemmed |
Estudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratos |
title_sort |
Estudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratos |
author |
Oliveira, Claudio Ricardo de |
author_facet |
Oliveira, Claudio Ricardo de |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/5091705745613726 |
dc.contributor.author.fl_str_mv |
Oliveira, Claudio Ricardo de |
dc.contributor.advisor1.fl_str_mv |
Araújo, Heloísa Sobreiro Selistre de |
dc.contributor.advisor1Lattes.fl_str_mv |
http://genos.cnpq.br:12010/dwlattes/owa/prc_imp_cv_int?f_cod=K4787059Y2 |
dc.contributor.authorID.fl_str_mv |
5f4304d8-aa9e-4739-9956-4db5e4d5b70c |
contributor_str_mv |
Araújo, Heloísa Sobreiro Selistre de |
dc.subject.por.fl_str_mv |
Hérnia ventral Abdomen DisBa-01 Cicatrização de feridas Ratos |
topic |
Hérnia ventral Abdomen DisBa-01 Cicatrização de feridas Ratos Ventral hernial Abdomen DisBa-01 Wound healing Rats CIENCIAS BIOLOGICAS::FISIOLOGIA |
dc.subject.eng.fl_str_mv |
Ventral hernial Abdomen DisBa-01 Wound healing Rats |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::FISIOLOGIA |
description |
INTRODUCTION: The incisional hernia (IH) is one of the most frequent complications after laparotomy, causing high rates of re-operations. Advances in suture material, in the guidelines for incisions, in the techniques of closure and use of dentures (screens), have failed in the elimination of this surgical complication. In this context, advances in the understanding of biological and structural changes of collagen may offer a new strategy in the treatment of IH, since the matrix disorders that induces the formation of primary hernia probably also affect the wound healing in laparotomies. OBJECTIVE: To investigate the effect of a recombinant desintegrin (DisBa-01) on the abdominal muscle healing process with incisional hernia (IH) regarding the fibroblasts and vessels proliferation and matrix metalopeptidase activity as well. The role of α2 and anti- αvβ3 integrins were investigated by antibody competition. METHODS: IH was induced by sectioning of the midline in 20 Wistar rats, divided into two groups: control that received the topical application of phosphate buffered saline (PBS) and DisBa-01 protein (0.5mg/Kg) dissolved in PBS. Another 10 rats were divided in two groups: one received anti-αVβ3 antibodies and the second anti-α2 subunit antibodies. At the end of 14 days, the animals were sacrificed and muscle fragments were stored in liquid nitrogen for determination of MMPs by zymography. The remaining fragments of the control and DisBa-01 groups were fixed in 10% formol in PBS (v/v). Blood vessels density, mononuclear cells, fibroblasts and collagen fibers were studied by optical microscopy in blue reticuline and Masson-trichrome. Data was analyzed by parametric tests were applied with critical level of 5%. RESULTS: All animals after surgical induction developed IH. However, after 14 days the persistence of the opened hernial ring in the controls but not in the DisBa-01 group. This protein induced the formation of a membrane that occluded the entire hernial ring. A significant increase in the number of mononuclear cells, fibroblasts, new vessels and MMP-2 activity was observed in DisBa-01 group (p<0.05). DisBa-01 group significant increased the collagen content. Anti-αVβ3 integrin antibodies produced similar results but not anti-α2 integrin blocking antibodies. CONCLUSION: These results indicate that DisBa-01 promoted vascular and fibroblastic cell proliferation and monocytic, and matrix remodeling. In addition, these results strongly indicate that αVβ3 integrin has an important role in the control of would healing and the blocking of this integrin may be an interesting therapeutical strategy in tissue repair conditions, including IH. |
publishDate |
2009 |
dc.date.available.fl_str_mv |
2009-08-27 2016-06-02T19:22:51Z |
dc.date.issued.fl_str_mv |
2009-04-29 |
dc.date.accessioned.fl_str_mv |
2016-06-02T19:22:51Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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OLIVEIRA, Claudio Ricardo de. Estudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratos. 2009. 92 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de São Carlos, São Carlos, 2009. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/1310 |
identifier_str_mv |
OLIVEIRA, Claudio Ricardo de. Estudo dos efeitos biológicos de uma desintegrina recombinante do veneno da serpente Bothrops alternatus, DisBa-01, sobre a musculatura da parede abdominal após indução de hérnia incisional em ratos. 2009. 92 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de São Carlos, São Carlos, 2009. |
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Universidade Federal de São Carlos |
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