Resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular

Detalhes bibliográficos
Autor(a) principal: Renner, Adriana Frias
Data de Publicação: 2010
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFSCAR
Texto Completo: https://repositorio.ufscar.br/handle/ufscar/5116
Resumo: The function of articular cartilage depends on the chondrocytes and on the components of the extracellular matrix, which in turn may be regulated by mechanical stimuli. Thus, changes in load support may affect its composition or its structure and interfere with their functional ability to sustain and distribute loads and minimize the stresses of contact. Thus, investigations of articular cartilage components, such as chondrocyte and or matrix components are essential for prevention and treatment of arthritic disease. A greater understanding of the relationship of use / disuse and degeneration as well as the consequences of situations such as shear stress, static load or unloading can generate in this tissue. The aim of this study was to evaluate the response of chondrocytes, proteoglycan, collagen and fibronectin in articular cartilage after application of a protocol of immobilization, stretching and joint remobilization. Material and Methods: We used 36 animals divided into six groups (n = 6): immobilized (I), immobilized and stretched seven days per week (IS7), immobilized and stretched three days per week (IS3), stretched seven days per week (S7), stretched three days per week (S3) and control (C). Groups I, IS3 and AS7 underwent four weeks of immobilization of the left hind limbs. Groups IS7 and IS3, after immobilization, were subjected to three weeks of the posterior muscle stretching of the left hind leg daily or three times per week, respectively. The S3 and S7 groups remained free in the cage for 4 weeks and subsequently underwent three weeks of posterior muscle stretching of the left hind limb daily or three times per week, respectively. After these procedures, the left ankle were collected, decalcified, processed in paraffin and stained with H&E, Safranin-O, Picrossiruius Red and immunostained with fibronectin and chondroitin sulfate 4 for further analysis. Two observers evaluated parameters such as chondrocyte cloning, loss of proteoglycan content, thin and thick fibrils collagen content, intensity of staining for fibronectin and chondroitin sulfate 4. For statistical analysis we used the following tests: Kruskal Wallis and post hoc Newman Keuls: cloning and the proteoglycan content of the different groups); Duncan multiple comparison: morphometric evaluation of cellularity; ANOVA and post hoc Tukey: proportion of thin and thick fibrils of collagen. For analysis of the immunohistochemistry reactions of fibronectin and chondroitin sulfate 4 it was used nonparametric test Kruscal Wallis and post hoc Newman Keuls. In all tests the significance level was p ≤ 0.05. Results: With respect to the cellularity IS7 group showed significant increase in cellularity compared to groups I and C. The IS3 group also showed significant celullar change with the formation of chondrocyte cloning compared to groups S7, S3 and C. The most significant loss of proteoglycan was in IS7 group compared to all other groups. The I group also lost significantly more proteoglycan than the others, except for IS7 group. With respect to collagen fibrils was observed that immobilization (I) significantly reduced the thin fibrils in relation to groups IS3, S7, S3 and C. The quantity of thick fibrils was influenced by mechanical overload, as there was a significant decrease of it in all groups compared to control. With respect to the findings of the fibronectin, the groups immobilized and stretched (IS3 and IS7) had significantly higher intensity staining of fibronectin than other groups. There was no statistical difference of chondroitin sulfate 4 immunostaining among the different groups. Conclusion: The protocols of muscle stretching after immobilization, applied on alternate days and daily provoked distinct adaptive responses in articular cartilage. The immobilization stimulated tissue atrophy that when stimulated by muscle stretching on alternate days, kept some matrix components, such as fine fibrils of collagen and proteoglycan, unlike the protocol used daily. Thus we can conclude that muscle stretching applied in previously immobilized joints should be applied with caution, on alternate days of mechanical stimulation.
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spelling Renner, Adriana FriasRosa, Stela Márcia Mattiello Gonçalveshttp://lattes.cnpq.br/1406279816228350http://lattes.cnpq.br/7079740194427120e8166c11-4f74-4291-a02d-49b8465752702016-06-02T20:18:12Z2010-06-082016-06-02T20:18:12Z2010-03-29RENNER, Adriana Frias. Resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular. 2010. 113 f. Tese (Doutorado em Ciências Biológicas) - Universidade Federal de São Carlos, São Carlos, 2010.https://repositorio.ufscar.br/handle/ufscar/5116The function of articular cartilage depends on the chondrocytes and on the components of the extracellular matrix, which in turn may be regulated by mechanical stimuli. Thus, changes in load support may affect its composition or its structure and interfere with their functional ability to sustain and distribute loads and minimize the stresses of contact. Thus, investigations of articular cartilage components, such as chondrocyte and or matrix components are essential for prevention and treatment of arthritic disease. A greater understanding of the relationship of use / disuse and degeneration as well as the consequences of situations such as shear stress, static load or unloading can generate in this tissue. The aim of this study was to evaluate the response of chondrocytes, proteoglycan, collagen and fibronectin in articular cartilage after application of a protocol of immobilization, stretching and joint remobilization. Material and Methods: We used 36 animals divided into six groups (n = 6): immobilized (I), immobilized and stretched seven days per week (IS7), immobilized and stretched three days per week (IS3), stretched seven days per week (S7), stretched three days per week (S3) and control (C). Groups I, IS3 and AS7 underwent four weeks of immobilization of the left hind limbs. Groups IS7 and IS3, after immobilization, were subjected to three weeks of the posterior muscle stretching of the left hind leg daily or three times per week, respectively. The S3 and S7 groups remained free in the cage for 4 weeks and subsequently underwent three weeks of posterior muscle stretching of the left hind limb daily or three times per week, respectively. After these procedures, the left ankle were collected, decalcified, processed in paraffin and stained with H&E, Safranin-O, Picrossiruius Red and immunostained with fibronectin and chondroitin sulfate 4 for further analysis. Two observers evaluated parameters such as chondrocyte cloning, loss of proteoglycan content, thin and thick fibrils collagen content, intensity of staining for fibronectin and chondroitin sulfate 4. For statistical analysis we used the following tests: Kruskal Wallis and post hoc Newman Keuls: cloning and the proteoglycan content of the different groups); Duncan multiple comparison: morphometric evaluation of cellularity; ANOVA and post hoc Tukey: proportion of thin and thick fibrils of collagen. For analysis of the immunohistochemistry reactions of fibronectin and chondroitin sulfate 4 it was used nonparametric test Kruscal Wallis and post hoc Newman Keuls. In all tests the significance level was p ≤ 0.05. Results: With respect to the cellularity IS7 group showed significant increase in cellularity compared to groups I and C. The IS3 group also showed significant celullar change with the formation of chondrocyte cloning compared to groups S7, S3 and C. The most significant loss of proteoglycan was in IS7 group compared to all other groups. The I group also lost significantly more proteoglycan than the others, except for IS7 group. With respect to collagen fibrils was observed that immobilization (I) significantly reduced the thin fibrils in relation to groups IS3, S7, S3 and C. The quantity of thick fibrils was influenced by mechanical overload, as there was a significant decrease of it in all groups compared to control. With respect to the findings of the fibronectin, the groups immobilized and stretched (IS3 and IS7) had significantly higher intensity staining of fibronectin than other groups. There was no statistical difference of chondroitin sulfate 4 immunostaining among the different groups. Conclusion: The protocols of muscle stretching after immobilization, applied on alternate days and daily provoked distinct adaptive responses in articular cartilage. The immobilization stimulated tissue atrophy that when stimulated by muscle stretching on alternate days, kept some matrix components, such as fine fibrils of collagen and proteoglycan, unlike the protocol used daily. Thus we can conclude that muscle stretching applied in previously immobilized joints should be applied with caution, on alternate days of mechanical stimulation.A função da cartilagem articular é dependente do condrócito e dos componentes de sua matriz extracelular, que por sua vez, podem ser regulados por estímulos mecânicos. Assim, alterações no suporte de carga podem afetar sua composição ou sua estrutura e interferir na sua capacidade funcional de sustentar e distribuir cargas e minimizar os estresses de contato. Desta forma, investigações dos componentes da cartilagem articular, como o condrócito e ou componentes da matriz são essenciais para prevenção e tratamento de doenças articulares. É necessário um maior entendimento das relações de uso/desuso e degeneração, assim como das conseqüências que situações como estresse de cisalhamento, carga estática prolongada ou ausência de carga possam gerar neste tecido. O objetivo do presente estudo foi avaliar a resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular. Material e Métodos: foram utilizados 36 animais divididos em 6 grupos (n=6): imobilizado (I), imobilizado e alongado 7 dias por semana (IA7), imobilizado e alongado 3 dias por semana (IA3), alongado 7 dias por semana (A7), alongado 3 dias por semana (A3), e controle (C). Os grupos I, IA7 e IA3 foram submetidos a 4 semanas de imobilização da pata traseira esquerda. Os grupos IA7 e IA3, após a imobilização, foram submetidos a 3 semanas de alongamento da musculatura posterior da pata traseira esquerda diariamente ou 3 vezes por semana, respectivamente. Os grupos A7 e A3 permaneceram livres na gaiola por 4 semanas e posteriormente foram submetidos a 3 semanas de alongamento da musculatura posterior da pata traseira esquerda diariamente ou em dias alternados, respectivamente. Após esses procedimentos, os tornozelos esquerdos foram coletados, descalcificados, processados em parafina e corados com H&E, Safranina, Picrossiruius Red e imunomarcados para fibronectina e sulfato de condroitina 4 para posterior análise. Foram avaliados por dois observadores parâmetros como: celularidade, contagem de clones, perda de proteoglicanos, conteúdo de fibrilas finas e grossas de colágeno e expressão de fibronectina e sulfato de condroitina 4. Para comparação destes parâmetros entre os diferentes grupos foram utilizados os seguintes testes estatísticos: Kruskal Wallis com post hoc Newman Keuls: formação de clones e conteúdo de proteoglicanas; Comparações múltiplas de Duncan: avaliação morfométrica de celularidade e Anova com post hoc de Tukey: proporção das fibrilas finas e grossas de colágeno. Para análise das reações de imunohistoquímica para fibronectina e sulfato de condroitina 4 foi utilizado o teste não paramétrico de Kruscal Wallis e post hoc Newman Keuls. Em todos os testes o nível de significância foi de p≤0,05. Resultados: com relação a celularidade o grupo IA7 apresentou aumento significativo da celularidade em relação aos grupos I e C. O grupo IA3 também apresentou alteração celular significativa com formação de clones em relação aos grupos A7, A3 e C. A maior perda significativa de proteoglicanas foi do grupo IA7 em relação a todos os outros grupos. O grupo I também perdeu significativamente mais proteoglicanas que os demais, somente não com relação ao grupo IA7. Com relação às fibrilas colágenas foi observado que a imobilização (I) reduziu significativamente as fibrilas finas em relação aos grupos IA3, A7, A3 e C. Já a quantidade de fibrilas grossas sofreu influência da sobrecarga mecânica, pois que houve diminuição significativa das mesmas em todos os grupos em relação ao controle. Com relação aos achados de fibronectina, os grupos imobilizados e alongados (IA7 e IA3) apresentaram significativamente maior intensidade de marcação desta que os outros grupos. Não houve diferença estatística das imunomarcações para sulfato de condroitina 4 entre os diferentes grupos. Conclusão: Os protocolos de alongamento muscular, após imobilização, realizados em dias alternados e diariamente, provocaram respostas adaptativas distintas na cartilagem articular. A imobilização desencadeou um quadro de atrofia tecidual que quando estimulada por alongamentos musculares em dias alternados, manteve alguns componentes da matriz, como fibrilas finas de colágeno e proteoglicana. Esta resposta foi agravada quando o mesmo protocolo foi aplicado diariamente. Desta forma, podemos concluir que o alongamento muscular aplicado em articulações previamente imobilizadas deve ser aplicado com cautela, respeitando períodos intercalados de estímulo mecânico.Universidade Federal de Sao Carlosapplication/pdfporUniversidade Federal de São CarlosPrograma de Pós-Graduação em Fisioterapia - PPGFtUFSCarBRFisioterapiaCartilagem articularImobilização articularAlongamento muscularColágenoProteoglicanaFibronectinaArticular cartilageCollagenProteoglycanFibronectinPhysical therapyCIENCIAS DA SAUDE::FISIOTERAPIA E TERAPIA OCUPACIONALResposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articularinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis-1-14f936219-f7d7-49fc-9dcc-ce3f39472005info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINAL3010.pdfapplication/pdf1781268https://repositorio.ufscar.br/bitstream/ufscar/5116/1/3010.pdf812b3af521e82fd881dcbddfcb1454b8MD51THUMBNAIL3010.pdf.jpg3010.pdf.jpgIM Thumbnailimage/jpeg7334https://repositorio.ufscar.br/bitstream/ufscar/5116/2/3010.pdf.jpg721af2efe70723c93a1bf9608067cc33MD52ufscar/51162023-09-18 18:31:05.592oai:repositorio.ufscar.br:ufscar/5116Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:05Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false
dc.title.por.fl_str_mv Resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular
title Resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular
spellingShingle Resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular
Renner, Adriana Frias
Fisioterapia
Cartilagem articular
Imobilização articular
Alongamento muscular
Colágeno
Proteoglicana
Fibronectina
Articular cartilage
Collagen
Proteoglycan
Fibronectin
Physical therapy
CIENCIAS DA SAUDE::FISIOTERAPIA E TERAPIA OCUPACIONAL
title_short Resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular
title_full Resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular
title_fullStr Resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular
title_full_unstemmed Resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular
title_sort Resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular
author Renner, Adriana Frias
author_facet Renner, Adriana Frias
author_role author
dc.contributor.authorlattes.por.fl_str_mv http://lattes.cnpq.br/7079740194427120
dc.contributor.author.fl_str_mv Renner, Adriana Frias
dc.contributor.advisor1.fl_str_mv Rosa, Stela Márcia Mattiello Gonçalves
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/1406279816228350
dc.contributor.authorID.fl_str_mv e8166c11-4f74-4291-a02d-49b846575270
contributor_str_mv Rosa, Stela Márcia Mattiello Gonçalves
dc.subject.por.fl_str_mv Fisioterapia
Cartilagem articular
Imobilização articular
Alongamento muscular
Colágeno
Proteoglicana
Fibronectina
topic Fisioterapia
Cartilagem articular
Imobilização articular
Alongamento muscular
Colágeno
Proteoglicana
Fibronectina
Articular cartilage
Collagen
Proteoglycan
Fibronectin
Physical therapy
CIENCIAS DA SAUDE::FISIOTERAPIA E TERAPIA OCUPACIONAL
dc.subject.eng.fl_str_mv Articular cartilage
Collagen
Proteoglycan
Fibronectin
Physical therapy
dc.subject.cnpq.fl_str_mv CIENCIAS DA SAUDE::FISIOTERAPIA E TERAPIA OCUPACIONAL
description The function of articular cartilage depends on the chondrocytes and on the components of the extracellular matrix, which in turn may be regulated by mechanical stimuli. Thus, changes in load support may affect its composition or its structure and interfere with their functional ability to sustain and distribute loads and minimize the stresses of contact. Thus, investigations of articular cartilage components, such as chondrocyte and or matrix components are essential for prevention and treatment of arthritic disease. A greater understanding of the relationship of use / disuse and degeneration as well as the consequences of situations such as shear stress, static load or unloading can generate in this tissue. The aim of this study was to evaluate the response of chondrocytes, proteoglycan, collagen and fibronectin in articular cartilage after application of a protocol of immobilization, stretching and joint remobilization. Material and Methods: We used 36 animals divided into six groups (n = 6): immobilized (I), immobilized and stretched seven days per week (IS7), immobilized and stretched three days per week (IS3), stretched seven days per week (S7), stretched three days per week (S3) and control (C). Groups I, IS3 and AS7 underwent four weeks of immobilization of the left hind limbs. Groups IS7 and IS3, after immobilization, were subjected to three weeks of the posterior muscle stretching of the left hind leg daily or three times per week, respectively. The S3 and S7 groups remained free in the cage for 4 weeks and subsequently underwent three weeks of posterior muscle stretching of the left hind limb daily or three times per week, respectively. After these procedures, the left ankle were collected, decalcified, processed in paraffin and stained with H&E, Safranin-O, Picrossiruius Red and immunostained with fibronectin and chondroitin sulfate 4 for further analysis. Two observers evaluated parameters such as chondrocyte cloning, loss of proteoglycan content, thin and thick fibrils collagen content, intensity of staining for fibronectin and chondroitin sulfate 4. For statistical analysis we used the following tests: Kruskal Wallis and post hoc Newman Keuls: cloning and the proteoglycan content of the different groups); Duncan multiple comparison: morphometric evaluation of cellularity; ANOVA and post hoc Tukey: proportion of thin and thick fibrils of collagen. For analysis of the immunohistochemistry reactions of fibronectin and chondroitin sulfate 4 it was used nonparametric test Kruscal Wallis and post hoc Newman Keuls. In all tests the significance level was p ≤ 0.05. Results: With respect to the cellularity IS7 group showed significant increase in cellularity compared to groups I and C. The IS3 group also showed significant celullar change with the formation of chondrocyte cloning compared to groups S7, S3 and C. The most significant loss of proteoglycan was in IS7 group compared to all other groups. The I group also lost significantly more proteoglycan than the others, except for IS7 group. With respect to collagen fibrils was observed that immobilization (I) significantly reduced the thin fibrils in relation to groups IS3, S7, S3 and C. The quantity of thick fibrils was influenced by mechanical overload, as there was a significant decrease of it in all groups compared to control. With respect to the findings of the fibronectin, the groups immobilized and stretched (IS3 and IS7) had significantly higher intensity staining of fibronectin than other groups. There was no statistical difference of chondroitin sulfate 4 immunostaining among the different groups. Conclusion: The protocols of muscle stretching after immobilization, applied on alternate days and daily provoked distinct adaptive responses in articular cartilage. The immobilization stimulated tissue atrophy that when stimulated by muscle stretching on alternate days, kept some matrix components, such as fine fibrils of collagen and proteoglycan, unlike the protocol used daily. Thus we can conclude that muscle stretching applied in previously immobilized joints should be applied with caution, on alternate days of mechanical stimulation.
publishDate 2010
dc.date.available.fl_str_mv 2010-06-08
2016-06-02T20:18:12Z
dc.date.issued.fl_str_mv 2010-03-29
dc.date.accessioned.fl_str_mv 2016-06-02T20:18:12Z
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dc.identifier.citation.fl_str_mv RENNER, Adriana Frias. Resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular. 2010. 113 f. Tese (Doutorado em Ciências Biológicas) - Universidade Federal de São Carlos, São Carlos, 2010.
dc.identifier.uri.fl_str_mv https://repositorio.ufscar.br/handle/ufscar/5116
identifier_str_mv RENNER, Adriana Frias. Resposta do condrócito, proteoglicana, colágeno e fibronectina da cartilagem articular, após aplicação de um protocolo de imobilização, alongamento e remobilização articular. 2010. 113 f. Tese (Doutorado em Ciências Biológicas) - Universidade Federal de São Carlos, São Carlos, 2010.
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