Condrotoxicidade e efeitos inflamatórios in vitro e in vivo da ropivacaína em equinos
Autor(a) principal: | |
---|---|
Data de Publicação: | 2019 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional Manancial UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/16509 |
Resumo: | Local anesthetics are routinely used in lameness diagnosis and joint pain control in horses, but because of their potential toxicity, they should be used with caution in order to avoid the development of joint disease. Previous studies in humans demonstrate ropivacaine as a less toxic alternative, motivating interest in the use of this anesthetic by the intra-articular (IA) route in horses. The objective of this study was to evaluate the IA effects of ropivacaine in vitro and in vivo. For viability evaluation, monolayer chondrocytes were divided into six groups and exposed to 30 minutes of treatment. The groups were: chondrogenic medium (DMEM), ropivacaine 7.5mg/ml, ropivacaine 10mg/ml, saline 0.9%, mepivacaine 20mg/ml and mepivacaine 30mg/ml. Chondrocyte viability was assessed by trypan blue, MTT and flow cytometry methods. To evaluate gene expression in vitro, pellets cultures of chondrocytes were used and submitted to 30 minutes of the following treatments: mepivacaine 20mg/ml, ropivacaine 10mg/ml and saline 0.9%. The cells were evaluated by Real-Time Quantitative Polymerase Chain Reaction (qPCR) to investigate the expression of Ilb1, Il6, Mmp9, Mmp13, Acan and Col2a1. The in vivo study was performed with the administration of 10 mg/ml ropivacaine and saline in the tibiotarsal joints of eight ponies. Synovial fluid samples were collected before and 24 hours after treatment. Then, samples were evaluated for total nucleated cells count, total protein and mucin precipitate evaluation. After 24 hours, the ponies were anesthetized and samples of synovia and cartilage were collected through arthroscopy to evaluate by qPCR. In the synovia and cartilage samples Ilb1, Il6, Mmp9 and Mmp13 were evaluated; genes Acan and Col2a1 were also evaluated in the cartilage samples. The evaluation of cell viability produced different results in the different assays: in the trypan blue method there was no difference; the MTT method demonstrated less viability of cells treated with ropivacaine than DMEM. Differently, flow cytometry did not show differences between ropivacaine 7.5mg ml and DMEM. In chondrocytes cultured in pellets, the expression of the evaluated genes was similar in the mepivacaine, ropivacaine and saline treatments. In pellets treated with mepivacaine and ropivacaine there was downregulation of the expression of Il6 and Mmp13 before and after treatment. After exposure to ropivacaine, there was downregulation of Mmp9 levels. In turn, the short exposure of the pellets to mepivacaine caused an increase in Col2a1 expression. In vivo results demonstrated that ropivacaine and saline cause inflammation in the synovial fluid when applied IA. However, no differences were observed between anabolic and cartilage catabolic markers between treatments. Only Mmp9 levels in synovia were increased following ropivacaine treatment. The in vitro results of this study suggest that short exposure to ropivacaine may result in a lower cell death rate than exposure to mepivacaine; in addition, mepivacaine and ropivacaine did not cause unbalance of chondrocyte homeostasis and appear to have some anti-inflammatory effect. Finally, the in vivo study revealed similarity between the effects of the IA treatment of ropivacaine and saline, suggesting that this anesthetic is a safe option for IA use. |
id |
UFSM-20_56ae5e45322c1d4d0f8a4f0f3adc6e82 |
---|---|
oai_identifier_str |
oai:repositorio.ufsm.br:1/16509 |
network_acronym_str |
UFSM-20 |
network_name_str |
Repositório Institucional Manancial UFSM |
repository_id_str |
3913 |
spelling |
2019-05-09T19:26:52Z2019-05-09T19:26:52Z2019-03-07http://repositorio.ufsm.br/handle/1/16509Local anesthetics are routinely used in lameness diagnosis and joint pain control in horses, but because of their potential toxicity, they should be used with caution in order to avoid the development of joint disease. Previous studies in humans demonstrate ropivacaine as a less toxic alternative, motivating interest in the use of this anesthetic by the intra-articular (IA) route in horses. The objective of this study was to evaluate the IA effects of ropivacaine in vitro and in vivo. For viability evaluation, monolayer chondrocytes were divided into six groups and exposed to 30 minutes of treatment. The groups were: chondrogenic medium (DMEM), ropivacaine 7.5mg/ml, ropivacaine 10mg/ml, saline 0.9%, mepivacaine 20mg/ml and mepivacaine 30mg/ml. Chondrocyte viability was assessed by trypan blue, MTT and flow cytometry methods. To evaluate gene expression in vitro, pellets cultures of chondrocytes were used and submitted to 30 minutes of the following treatments: mepivacaine 20mg/ml, ropivacaine 10mg/ml and saline 0.9%. The cells were evaluated by Real-Time Quantitative Polymerase Chain Reaction (qPCR) to investigate the expression of Ilb1, Il6, Mmp9, Mmp13, Acan and Col2a1. The in vivo study was performed with the administration of 10 mg/ml ropivacaine and saline in the tibiotarsal joints of eight ponies. Synovial fluid samples were collected before and 24 hours after treatment. Then, samples were evaluated for total nucleated cells count, total protein and mucin precipitate evaluation. After 24 hours, the ponies were anesthetized and samples of synovia and cartilage were collected through arthroscopy to evaluate by qPCR. In the synovia and cartilage samples Ilb1, Il6, Mmp9 and Mmp13 were evaluated; genes Acan and Col2a1 were also evaluated in the cartilage samples. The evaluation of cell viability produced different results in the different assays: in the trypan blue method there was no difference; the MTT method demonstrated less viability of cells treated with ropivacaine than DMEM. Differently, flow cytometry did not show differences between ropivacaine 7.5mg ml and DMEM. In chondrocytes cultured in pellets, the expression of the evaluated genes was similar in the mepivacaine, ropivacaine and saline treatments. In pellets treated with mepivacaine and ropivacaine there was downregulation of the expression of Il6 and Mmp13 before and after treatment. After exposure to ropivacaine, there was downregulation of Mmp9 levels. In turn, the short exposure of the pellets to mepivacaine caused an increase in Col2a1 expression. In vivo results demonstrated that ropivacaine and saline cause inflammation in the synovial fluid when applied IA. However, no differences were observed between anabolic and cartilage catabolic markers between treatments. Only Mmp9 levels in synovia were increased following ropivacaine treatment. The in vitro results of this study suggest that short exposure to ropivacaine may result in a lower cell death rate than exposure to mepivacaine; in addition, mepivacaine and ropivacaine did not cause unbalance of chondrocyte homeostasis and appear to have some anti-inflammatory effect. Finally, the in vivo study revealed similarity between the effects of the IA treatment of ropivacaine and saline, suggesting that this anesthetic is a safe option for IA use.Os anestésicos locais são rotineiramente usados no diagnóstico de claudicações e controle da dor articular em equinos. Contudo, devido à sua potencial toxicidade, devem ser usados com cautela pelo risco de desenvolvimento de doença articular. Estudos prévios em humanos demonstram a ropivacaína como uma alternativa menos tóxica, motivando o interesse no uso deste anestésico pela via intraarticular (IA) em equinos. O objetivo deste trabalho foi avaliar os efeitos IA da ropivacaína em técnicas in vitro e in vivo. Para avaliação de viabilidade, condrócitos em monocamada foram divididos em seis grupos e expostos a 30 minutos dos seguintes tratamentos: meio condrogênico (DMEM), ropivacaína 7.5mg/ml, ropivacaína 10mg/ml, solução salina 0.9%, mepivacaína 20mg/ml e mepivacaína 30mg/ml. A viabilidade dos condrócitos foi avaliada pelos métodos de trypan blue, MTT e citometria de fluxo com fluoróforo iodeto de propídio. Para avaliação de expressão gênica in vitro foram utilizados cultivos de condrócitos em pellets submetidos a 30 minutos dos seguintes tratamentos: mepivacaína 20 mg/ml, ropivacaína 10mg/ml e solução salina 0.9%. As células foram avaliadas por Reação em Cadeia Polimerase Quantitativa em Tempo Real (qPCR) para investigar a expressão de Il1, Il6, Mmp9, Mmp13, Acan e Col2a1. O estudo in vivo foi realizado com a administração IA de ropivacaína 10mg/ml e solução salina nas articulações tibiotársicas de oito pôneis. Imediatamente antes da administração dos tratamentos e 24 horas depois foi realizada coleta de líquido sinovial. As amostras foram avaliadas quanto à contagem de células nucleadas, proteína total e qualidade da mucina. Sob anestesia geral, através de portais artroscópicos, foram coletadas amostras de sinóvia, para determinação de concentrações de Ibl1, Il6, Mmp9 e Mmp13, e cartilagem, para as mesmas determinações mais Acan e Col2a1 por qPCR. A avaliação de viabilidade celular produziu resultados distintos nos diferentes ensaios. No método de trypan blue não houve diferença; o método de MTT demonstrou menor viabilidade de células tratadas com ropivacaína que DMEM. Diferentemente, a citometria de fluxo não evidenciou diferenças entre ropivacaína 7.5mg/ml e DMEM. Em condrócitos cultivados em pellets, a expressão dos genes avaliados foi semelhante nos tratamentos com mepivacaina, ropivacaína e salina. Nos pellets tratados com mepivacaína e ropivacaína houve reduçãos da expressão de Il6 e Mmp13 antes e após o tratamento. Enquanto a exposição à ropivacaína suprimiu os níveis de Mmp9, A exposição à mepivacaína resultou em aumento na expressão de Col2a1. Os resultados in vivo demonstraram que ropivacaína e salina causam inflamação no líquido sinovial, observado pelo aumento de celularidade, quando aplicados via IA. Entretanto, não foram observadas diferenças entre marcadores anabólicos e catabólicos de cartilagem entre tratamentos. Apenas os níveis de Mmp9 na sinóvia foram aumentados após tratamento com ropivacaína. Os resultados in vitro deste trabalho sugerem que a curta exposição à ropivacaína pode resultar em menor taxa de morte celular que a exposição à mepivacaína; adicionalmente, a exposição à ropivacaína não causa perda da homeostase dos condrócitos. Por fim, o estudo in vivo revelou similaridade entre os efeitos do tratamento IA da ropivacaína e salina, sugerindo ser este anestésico uma opção segura para o uso IA.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESporUniversidade Federal de Santa MariaCentro de Ciências RuraisPrograma de Pós-Graduação em Medicina VeterináriaUFSMBrasilMedicina VeterináriaAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessCondrócitosAnestésicos locaisIntra-articularGeneChondrocytesLocal anestheticsIntra-articularCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIACondrotoxicidade e efeitos inflamatórios in vitro e in vivo da ropivacaína em equinosCondrotoxicity and inflammatory effects in vitro and in vivo of ropivacaine in equineinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisDe La Côrte, Flávio Desessardshttp://lattes.cnpq.br/4040388452531898Krause, Alexandrehttp://lattes.cnpq.br/7760558908777387Portela Junior, Valério Valdetar Marqueshttp://lattes.cnpq.br/0564876468635367Krause, Luciana Maria Fontanarihttp://lattes.cnpq.br/9844890896121847Faleiros, Rafael Resendehttp://lattes.cnpq.br/4660433855798183http://lattes.cnpq.br/2862227568895947Silva, Gabriele Biavaschi da5005000000076000a814134-4c3e-4a94-99ed-af7f683bf8a0a5c20d24-09f6-4eb0-9286-5508a4f31ae148b1731f-9fa9-4553-ab4a-18302b9fc514e40a48f8-b499-4599-ae06-9f609dbacadcdacc436c-09f7-4dd4-aabe-1d19ee49df45c254a1f7-f544-44fa-8d07-37a7caa3201dreponame:Repositório Institucional Manancial UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALTES_PPGMV_2019_SILVA_GABRIELE.pdfTES_PPGMV_2019_SILVA_GABRIELE.pdfTese de Doutoradoapplication/pdf1914368http://repositorio.ufsm.br/bitstream/1/16509/1/TES_PPGMV_2019_SILVA_GABRIELE.pdf983aa0ab98430328a7e4caa450744965MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8805http://repositorio.ufsm.br/bitstream/1/16509/2/license_rdf4460e5956bc1d1639be9ae6146a50347MD52LICENSElicense.txtlicense.txttext/plain; charset=utf-81956http://repositorio.ufsm.br/bitstream/1/16509/3/license.txt2f0571ecee68693bd5cd3f17c1e075dfMD53TEXTTES_PPGMV_2019_SILVA_GABRIELE.pdf.txtTES_PPGMV_2019_SILVA_GABRIELE.pdf.txtExtracted texttext/plain205963http://repositorio.ufsm.br/bitstream/1/16509/4/TES_PPGMV_2019_SILVA_GABRIELE.pdf.txtc33ea144becf3ae5ae7b993f0dd455b2MD54THUMBNAILTES_PPGMV_2019_SILVA_GABRIELE.pdf.jpgTES_PPGMV_2019_SILVA_GABRIELE.pdf.jpgIM Thumbnailimage/jpeg4690http://repositorio.ufsm.br/bitstream/1/16509/5/TES_PPGMV_2019_SILVA_GABRIELE.pdf.jpg55a0b298c64955cc9beb2ac206620e05MD551/165092022-05-06 16:55:01.17oai:repositorio.ufsm.br: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ório Institucionalhttp://repositorio.ufsm.br/PUBhttp://repositorio.ufsm.br/oai/requestouvidoria@ufsm.bropendoar:39132022-05-06T19:55:01Repositório Institucional Manancial UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.por.fl_str_mv |
Condrotoxicidade e efeitos inflamatórios in vitro e in vivo da ropivacaína em equinos |
dc.title.alternative.eng.fl_str_mv |
Condrotoxicity and inflammatory effects in vitro and in vivo of ropivacaine in equine |
title |
Condrotoxicidade e efeitos inflamatórios in vitro e in vivo da ropivacaína em equinos |
spellingShingle |
Condrotoxicidade e efeitos inflamatórios in vitro e in vivo da ropivacaína em equinos Silva, Gabriele Biavaschi da Condrócitos Anestésicos locais Intra-articular Gene Chondrocytes Local anesthetics Intra-articular CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
title_short |
Condrotoxicidade e efeitos inflamatórios in vitro e in vivo da ropivacaína em equinos |
title_full |
Condrotoxicidade e efeitos inflamatórios in vitro e in vivo da ropivacaína em equinos |
title_fullStr |
Condrotoxicidade e efeitos inflamatórios in vitro e in vivo da ropivacaína em equinos |
title_full_unstemmed |
Condrotoxicidade e efeitos inflamatórios in vitro e in vivo da ropivacaína em equinos |
title_sort |
Condrotoxicidade e efeitos inflamatórios in vitro e in vivo da ropivacaína em equinos |
author |
Silva, Gabriele Biavaschi da |
author_facet |
Silva, Gabriele Biavaschi da |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
De La Côrte, Flávio Desessards |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/4040388452531898 |
dc.contributor.referee1.fl_str_mv |
Krause, Alexandre |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/7760558908777387 |
dc.contributor.referee2.fl_str_mv |
Portela Junior, Valério Valdetar Marques |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/0564876468635367 |
dc.contributor.referee3.fl_str_mv |
Krause, Luciana Maria Fontanari |
dc.contributor.referee3Lattes.fl_str_mv |
http://lattes.cnpq.br/9844890896121847 |
dc.contributor.referee4.fl_str_mv |
Faleiros, Rafael Resende |
dc.contributor.referee4Lattes.fl_str_mv |
http://lattes.cnpq.br/4660433855798183 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/2862227568895947 |
dc.contributor.author.fl_str_mv |
Silva, Gabriele Biavaschi da |
contributor_str_mv |
De La Côrte, Flávio Desessards Krause, Alexandre Portela Junior, Valério Valdetar Marques Krause, Luciana Maria Fontanari Faleiros, Rafael Resende |
dc.subject.por.fl_str_mv |
Condrócitos Anestésicos locais Intra-articular Gene |
topic |
Condrócitos Anestésicos locais Intra-articular Gene Chondrocytes Local anesthetics Intra-articular CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
dc.subject.eng.fl_str_mv |
Chondrocytes Local anesthetics Intra-articular |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
description |
Local anesthetics are routinely used in lameness diagnosis and joint pain control in horses, but because of their potential toxicity, they should be used with caution in order to avoid the development of joint disease. Previous studies in humans demonstrate ropivacaine as a less toxic alternative, motivating interest in the use of this anesthetic by the intra-articular (IA) route in horses. The objective of this study was to evaluate the IA effects of ropivacaine in vitro and in vivo. For viability evaluation, monolayer chondrocytes were divided into six groups and exposed to 30 minutes of treatment. The groups were: chondrogenic medium (DMEM), ropivacaine 7.5mg/ml, ropivacaine 10mg/ml, saline 0.9%, mepivacaine 20mg/ml and mepivacaine 30mg/ml. Chondrocyte viability was assessed by trypan blue, MTT and flow cytometry methods. To evaluate gene expression in vitro, pellets cultures of chondrocytes were used and submitted to 30 minutes of the following treatments: mepivacaine 20mg/ml, ropivacaine 10mg/ml and saline 0.9%. The cells were evaluated by Real-Time Quantitative Polymerase Chain Reaction (qPCR) to investigate the expression of Ilb1, Il6, Mmp9, Mmp13, Acan and Col2a1. The in vivo study was performed with the administration of 10 mg/ml ropivacaine and saline in the tibiotarsal joints of eight ponies. Synovial fluid samples were collected before and 24 hours after treatment. Then, samples were evaluated for total nucleated cells count, total protein and mucin precipitate evaluation. After 24 hours, the ponies were anesthetized and samples of synovia and cartilage were collected through arthroscopy to evaluate by qPCR. In the synovia and cartilage samples Ilb1, Il6, Mmp9 and Mmp13 were evaluated; genes Acan and Col2a1 were also evaluated in the cartilage samples. The evaluation of cell viability produced different results in the different assays: in the trypan blue method there was no difference; the MTT method demonstrated less viability of cells treated with ropivacaine than DMEM. Differently, flow cytometry did not show differences between ropivacaine 7.5mg ml and DMEM. In chondrocytes cultured in pellets, the expression of the evaluated genes was similar in the mepivacaine, ropivacaine and saline treatments. In pellets treated with mepivacaine and ropivacaine there was downregulation of the expression of Il6 and Mmp13 before and after treatment. After exposure to ropivacaine, there was downregulation of Mmp9 levels. In turn, the short exposure of the pellets to mepivacaine caused an increase in Col2a1 expression. In vivo results demonstrated that ropivacaine and saline cause inflammation in the synovial fluid when applied IA. However, no differences were observed between anabolic and cartilage catabolic markers between treatments. Only Mmp9 levels in synovia were increased following ropivacaine treatment. The in vitro results of this study suggest that short exposure to ropivacaine may result in a lower cell death rate than exposure to mepivacaine; in addition, mepivacaine and ropivacaine did not cause unbalance of chondrocyte homeostasis and appear to have some anti-inflammatory effect. Finally, the in vivo study revealed similarity between the effects of the IA treatment of ropivacaine and saline, suggesting that this anesthetic is a safe option for IA use. |
publishDate |
2019 |
dc.date.accessioned.fl_str_mv |
2019-05-09T19:26:52Z |
dc.date.available.fl_str_mv |
2019-05-09T19:26:52Z |
dc.date.issued.fl_str_mv |
2019-03-07 |
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 |
http://repositorio.ufsm.br/handle/1/16509 |
url |
http://repositorio.ufsm.br/handle/1/16509 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.cnpq.fl_str_mv |
500500000007 |
dc.relation.confidence.fl_str_mv |
600 |
dc.relation.authority.fl_str_mv |
0a814134-4c3e-4a94-99ed-af7f683bf8a0 a5c20d24-09f6-4eb0-9286-5508a4f31ae1 48b1731f-9fa9-4553-ab4a-18302b9fc514 e40a48f8-b499-4599-ae06-9f609dbacadc dacc436c-09f7-4dd4-aabe-1d19ee49df45 c254a1f7-f544-44fa-8d07-37a7caa3201d |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Medicina Veterinária |
dc.publisher.initials.fl_str_mv |
UFSM |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Medicina Veterinária |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional Manancial 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 |
Repositório Institucional Manancial UFSM |
collection |
Repositório Institucional Manancial UFSM |
bitstream.url.fl_str_mv |
http://repositorio.ufsm.br/bitstream/1/16509/1/TES_PPGMV_2019_SILVA_GABRIELE.pdf http://repositorio.ufsm.br/bitstream/1/16509/2/license_rdf http://repositorio.ufsm.br/bitstream/1/16509/3/license.txt http://repositorio.ufsm.br/bitstream/1/16509/4/TES_PPGMV_2019_SILVA_GABRIELE.pdf.txt http://repositorio.ufsm.br/bitstream/1/16509/5/TES_PPGMV_2019_SILVA_GABRIELE.pdf.jpg |
bitstream.checksum.fl_str_mv |
983aa0ab98430328a7e4caa450744965 4460e5956bc1d1639be9ae6146a50347 2f0571ecee68693bd5cd3f17c1e075df c33ea144becf3ae5ae7b993f0dd455b2 55a0b298c64955cc9beb2ac206620e05 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositório Institucional Manancial UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
ouvidoria@ufsm.br |
_version_ |
1808854704364453888 |