Evaluation of the role of CCR2 and CCR5 receptors in macrophage migration and alveolar bone repair outcome in mice
Ano de defesa: | 2017 |
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Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | |
Tipo de documento: | Dissertação |
Tipo de acesso: | Acesso aberto |
Idioma: | eng |
Instituição de defesa: |
Biblioteca Digitais de Teses e Dissertações da USP
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Programa de Pós-Graduação: |
Não Informado pela instituição
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Departamento: |
Não Informado pela instituição
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País: |
Não Informado pela instituição
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Palavras-chave em Português: | |
Link de acesso: | https://www.teses.usp.br/teses/disponiveis/25/25149/tde-26112021-151719/ |
Resumo: | Macrophages play important roles in bone repair, including the control of immune response and inflammation, and regulating the transition between granulation tissue to osteogenesis and formation of new bone after an injury. In this context, chemokine receptors, such as CCR5 and CCR2, seems to be key players in the chemotaxis of monocytes/macrophages to sites of tissue injury. The objective of this project is investigate simultaneously the role of CCR2 and CCR5 receptors in the cell migration and its subsequent impact on the alveolar repair process in mice. Mice C57BL/6 WT and CCR5KO, eight weeks old, were submitted to extraction of the right upper incisor and distributed in groups (N=5) control and treated with the antagonist for CCR2 (RS504393, 2mg/kg/24h), in order to allow of the blockage analysis individually or simultaneously of the receptors. Samples were collected in the 0h, 7d, 14d and 21days post extraction periods, and analyzed by micro-computed tomography (CT), histological analyzes (histomorphometry, immunohistochemistry and birefringence analysis), as well as molecular analysis by means of PCRArray for quantification of different markers involved in the repair process. Immunohistochemical analysis shows that CCR2 and CCR5 receptor blockade did not significantly influence the migration of monocytes/macrophages to alveolar bone repair. A similar pattern can be observed in MicroCT and in microscopic analyzes that do not demonstrate major changes in the parameters representatative of bone healing. On the other hand, molecular analyzes demonstrated that the simultaneous inhibition of CCR2 and CCR5 (CCR5KOantiCCR2 group) resulted in a a significantly higher mRNA expression of extracellular matrix markers (COL2A1 and MMP9), some bone markers (DMP1 and RANKL), FGF1 as well as IL-6 expression and decreased expression of growth factors (TGFb1 and VEGF), as well as RUNX-2 and cytokines (IL-10 and TNF-) when compared to WT-C. Therefore, we conclude that, although CCR2 and CCR5 receptor blockade result in significant modulation of of growth factors, proinflammatory cytokines and osteoclastogenic factors expression, there are no significant differences in the control of macrophage migration as well as in the subsequent bone repair outcome. |
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Biblioteca Digital de Teses e Dissertações da USP |
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Evaluation of the role of CCR2 and CCR5 receptors in macrophage migration and alveolar bone repair outcome in miceAvaliação do papel dos receptores CCR2 e CCR5 na migração de macrófagos e no processo de reparo ósseo alveolar em camundongosAlvéolo dentalBone repairCCR2CCR2CCR5CCR5Dental socketMacrófagoMacrophageOsteoimunologiaOsteoinmulogyReparo ósseoMacrophages play important roles in bone repair, including the control of immune response and inflammation, and regulating the transition between granulation tissue to osteogenesis and formation of new bone after an injury. In this context, chemokine receptors, such as CCR5 and CCR2, seems to be key players in the chemotaxis of monocytes/macrophages to sites of tissue injury. The objective of this project is investigate simultaneously the role of CCR2 and CCR5 receptors in the cell migration and its subsequent impact on the alveolar repair process in mice. Mice C57BL/6 WT and CCR5KO, eight weeks old, were submitted to extraction of the right upper incisor and distributed in groups (N=5) control and treated with the antagonist for CCR2 (RS504393, 2mg/kg/24h), in order to allow of the blockage analysis individually or simultaneously of the receptors. Samples were collected in the 0h, 7d, 14d and 21days post extraction periods, and analyzed by micro-computed tomography (CT), histological analyzes (histomorphometry, immunohistochemistry and birefringence analysis), as well as molecular analysis by means of PCRArray for quantification of different markers involved in the repair process. Immunohistochemical analysis shows that CCR2 and CCR5 receptor blockade did not significantly influence the migration of monocytes/macrophages to alveolar bone repair. A similar pattern can be observed in MicroCT and in microscopic analyzes that do not demonstrate major changes in the parameters representatative of bone healing. On the other hand, molecular analyzes demonstrated that the simultaneous inhibition of CCR2 and CCR5 (CCR5KOantiCCR2 group) resulted in a a significantly higher mRNA expression of extracellular matrix markers (COL2A1 and MMP9), some bone markers (DMP1 and RANKL), FGF1 as well as IL-6 expression and decreased expression of growth factors (TGFb1 and VEGF), as well as RUNX-2 and cytokines (IL-10 and TNF-) when compared to WT-C. Therefore, we conclude that, although CCR2 and CCR5 receptor blockade result in significant modulation of of growth factors, proinflammatory cytokines and osteoclastogenic factors expression, there are no significant differences in the control of macrophage migration as well as in the subsequent bone repair outcome.Sabe-se que macrófagos exercem um importante papel no processo de reparo tecidual, e estão presentes inclusive no processo de reparo ósseo, possivelmente atuando no controle e ativação da resposta imune/inflamatória, bem como na transição do tecido de granulação para um processo de osteogênese e a neo-formação de um tecido ósseo após a lesão. Neste sentido, os receptores de quimiocinas CCR2 e CCR5 parecem ser os principais envolvidos na quimiotaxia de monócitos/macrófagos para locais de injúria tecidual. Dessa forma, o objetivo deste projeto é investigar simultaneamente o papel dos receptores CCR2 e CCR5 na migração celular e seu impacto subsequente no processo de reparo alveolar. Camundongos C57Bl/6 WT e CCR5KO, com oito semanas de idade, foram submetidos à extração do incisivo superior direito e distribuídos em grupos (N=5) controle e tratados com o antagonista para CCR2 (RS504393, 2mg/Kg/24h), de modo a possibilitar a análise do bloqueio individual ou simultâneo dos receptores. As amostras foram coletadas nos períodos de 0h, 7d, 14d e 21dias pós-exodontia, e analisadas através de microtomografia computadorizada (MicroCT), análises histológicas (histomorfometria, imunoistoquímica e análise de birrefringência), bem como análise molecular por meio do PCRArray para quantificação de diferentes marcadores envolvidos no processo de reparo. A análise imuno-histoquímica nos mostra que o bloqueio dos receptores CCR2 e CCR5 não influenciou significativamente à migração de monócitos/macrófagos em direção ao reparo ósseo alveolar. Um padrão semelhante pode ser observado nas análises MicroCT e microscópicas que não demonstram mudanças importantes nos parâmetros representativos da cicatrização óssea . Já a análise molecular, demonstrou que essa mesma dupla inibição de CCR2 e CCR5 (grupo CCR5KOantiCCR2) resultou em uma expressão de mRNA significativamente maior de marcadores de matriz extracelular (COL2A1 e MMP9), alguns marcadores ósseos (DMP1 e RANKL ), FGF1 e também na expressão de IL-6 e uma menor expressão dos fatores de crescimento (TGFb1 e VEGF), bem como RUNX-2 e e as citocinas (IL-10 e TNF-) quando comparado ao WT-C. Assim concluímos que, embora, que o bloqueio dos receptores CCR2 e CCR5 resulte em uma modulação significativa de fatores de crescimento, citocinas pró-inflamátorias e expressão de fatores osteogênicos, não há diferença significativas no controle de migração de macrófagos, bem como no subsequente resultado do reparo ósseo alveolar.Biblioteca Digitais de Teses e Dissertações da USPGarlet, Gustavo PompermaierFonseca, Angélica Cristina2017-12-13info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/25/25149/tde-26112021-151719/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPLiberar o conteúdo para acesso público.info:eu-repo/semantics/openAccesseng2024-08-16T22:53:02Zoai:teses.usp.br:tde-26112021-151719Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212024-08-16T22:53:02Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false |
dc.title.none.fl_str_mv |
Evaluation of the role of CCR2 and CCR5 receptors in macrophage migration and alveolar bone repair outcome in mice Avaliação do papel dos receptores CCR2 e CCR5 na migração de macrófagos e no processo de reparo ósseo alveolar em camundongos |
title |
Evaluation of the role of CCR2 and CCR5 receptors in macrophage migration and alveolar bone repair outcome in mice |
spellingShingle |
Evaluation of the role of CCR2 and CCR5 receptors in macrophage migration and alveolar bone repair outcome in mice Fonseca, Angélica Cristina Alvéolo dental Bone repair CCR2 CCR2 CCR5 CCR5 Dental socket Macrófago Macrophage Osteoimunologia Osteoinmulogy Reparo ósseo |
title_short |
Evaluation of the role of CCR2 and CCR5 receptors in macrophage migration and alveolar bone repair outcome in mice |
title_full |
Evaluation of the role of CCR2 and CCR5 receptors in macrophage migration and alveolar bone repair outcome in mice |
title_fullStr |
Evaluation of the role of CCR2 and CCR5 receptors in macrophage migration and alveolar bone repair outcome in mice |
title_full_unstemmed |
Evaluation of the role of CCR2 and CCR5 receptors in macrophage migration and alveolar bone repair outcome in mice |
title_sort |
Evaluation of the role of CCR2 and CCR5 receptors in macrophage migration and alveolar bone repair outcome in mice |
author |
Fonseca, Angélica Cristina |
author_facet |
Fonseca, Angélica Cristina |
author_role |
author |
dc.contributor.none.fl_str_mv |
Garlet, Gustavo Pompermaier |
dc.contributor.author.fl_str_mv |
Fonseca, Angélica Cristina |
dc.subject.por.fl_str_mv |
Alvéolo dental Bone repair CCR2 CCR2 CCR5 CCR5 Dental socket Macrófago Macrophage Osteoimunologia Osteoinmulogy Reparo ósseo |
topic |
Alvéolo dental Bone repair CCR2 CCR2 CCR5 CCR5 Dental socket Macrófago Macrophage Osteoimunologia Osteoinmulogy Reparo ósseo |
description |
Macrophages play important roles in bone repair, including the control of immune response and inflammation, and regulating the transition between granulation tissue to osteogenesis and formation of new bone after an injury. In this context, chemokine receptors, such as CCR5 and CCR2, seems to be key players in the chemotaxis of monocytes/macrophages to sites of tissue injury. The objective of this project is investigate simultaneously the role of CCR2 and CCR5 receptors in the cell migration and its subsequent impact on the alveolar repair process in mice. Mice C57BL/6 WT and CCR5KO, eight weeks old, were submitted to extraction of the right upper incisor and distributed in groups (N=5) control and treated with the antagonist for CCR2 (RS504393, 2mg/kg/24h), in order to allow of the blockage analysis individually or simultaneously of the receptors. Samples were collected in the 0h, 7d, 14d and 21days post extraction periods, and analyzed by micro-computed tomography (CT), histological analyzes (histomorphometry, immunohistochemistry and birefringence analysis), as well as molecular analysis by means of PCRArray for quantification of different markers involved in the repair process. Immunohistochemical analysis shows that CCR2 and CCR5 receptor blockade did not significantly influence the migration of monocytes/macrophages to alveolar bone repair. A similar pattern can be observed in MicroCT and in microscopic analyzes that do not demonstrate major changes in the parameters representatative of bone healing. On the other hand, molecular analyzes demonstrated that the simultaneous inhibition of CCR2 and CCR5 (CCR5KOantiCCR2 group) resulted in a a significantly higher mRNA expression of extracellular matrix markers (COL2A1 and MMP9), some bone markers (DMP1 and RANKL), FGF1 as well as IL-6 expression and decreased expression of growth factors (TGFb1 and VEGF), as well as RUNX-2 and cytokines (IL-10 and TNF-) when compared to WT-C. Therefore, we conclude that, although CCR2 and CCR5 receptor blockade result in significant modulation of of growth factors, proinflammatory cytokines and osteoclastogenic factors expression, there are no significant differences in the control of macrophage migration as well as in the subsequent bone repair outcome. |
publishDate |
2017 |
dc.date.none.fl_str_mv |
2017-12-13 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://www.teses.usp.br/teses/disponiveis/25/25149/tde-26112021-151719/ |
url |
https://www.teses.usp.br/teses/disponiveis/25/25149/tde-26112021-151719/ |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
|
dc.rights.driver.fl_str_mv |
Liberar o conteúdo para acesso público. info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Liberar o conteúdo para acesso público. |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.coverage.none.fl_str_mv |
|
dc.publisher.none.fl_str_mv |
Biblioteca Digitais de Teses e Dissertações da USP |
publisher.none.fl_str_mv |
Biblioteca Digitais de Teses e Dissertações da USP |
dc.source.none.fl_str_mv |
reponame:Biblioteca Digital de Teses e Dissertações da USP instname:Universidade de São Paulo (USP) instacron:USP |
instname_str |
Universidade de São Paulo (USP) |
instacron_str |
USP |
institution |
USP |
reponame_str |
Biblioteca Digital de Teses e Dissertações da USP |
collection |
Biblioteca Digital de Teses e Dissertações da USP |
repository.name.fl_str_mv |
Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP) |
repository.mail.fl_str_mv |
virginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.br |
_version_ |
1815258489820282880 |