Evaluation of the role of CCR2 and CCR5 receptors in macrophage migration and alveolar bone repair outcome in mice

Detalhes bibliográficos
Ano de defesa: 2017
Autor(a) principal: Fonseca, Angélica Cristina
Orientador(a): Não Informado pela instituição
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: eng
Instituição de defesa: Biblioteca Digitais de Teses e Dissertações da USP
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
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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spelling 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
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