Efeitos da terapia celular, com células tronco mesenquimais do tecido adiposo, e do infiltrado de plasma rico em plaquetas no tratamento da osteoartrite não responsiva ao tratamento medicamentoso

Detalhes bibliográficos
Autor(a) principal: Laynna de Carvalho Schweich
Data de Publicação: 2021
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFMS
Texto Completo: https://repositorio.ufms.br/handle/123456789/3707
Resumo: The increase in individuals with Osteoarthritis (OA) has generated an increase in public spending on monitoring and maintaining treatments, which have lasted for years and are still not resolvable. This study is the first cell therapy proposal for OA carried out in the Brazilian Public Health System, in the State of Mato Grosso do Sul. The objective of the work was divided into two stages: the laboratory which aimed to analyze the quality of adipose-derived stem cells (ADSCs ) cultured from the 1st to the 10th passage, and later the clinic, which in an unprecedented way analyzed and compared 4 types of interventions in patients with OA: platelet-rich plasma (PRP), ADSCs, ADSCs + PRP and the standard surgical procedure performed in our hospital. The analysis of the ADSCs passages was performed by immunophenotyping assay, cell differentiation, comet assay, cytological and molecular cell death assay, morphological analysis, apoptosis, membrane integrity and viability in the flow cytometer. The evaluation of the patients was carried out by applying the questionnaires of WOMAC, SF-36 and EVA, and by analyzing the synovial fluid (inflammatory cytokines, enzymatic, colorimetric and viscosity analysis). Regarding the quality of ADSCs, the trypsinization process carried out during cultivation did not interfere with the genomic stability of the cells. But as of the 6th pass, ADSCs started to modify its capacity for juxtaposed deposition and proliferation in monolayer, as well as increasing its size. Cell viability was significantly reduced only on the 10th passage, and it was proven that this was due to apoptosis due to changes in membrane permeability. The results of the questionnaires applied to the patients, showed a greater improvement in the scores of the analyzed domains (initial x final evaluation / by group and final values / between groups) in the ADSCs + PRP group, followed by the ADSCs and PRP group. In the evaluation of inflammatory cytokines, there was a significant reduction (initial x final / per group) of the IL-1b cytokine only in the ADSCs + PRP (40%) and ADSCs (31%) groups, of IL-6 in the ADSCs + PRP group ( 72%), IL-8 in the ADSCs + PRP (50%) and ADSCs (31%) groups, and TNF in the ADSCs + PRP group (46%). Only the reductions in the ADSCs + PRP group were considered significant in the analysis of the final values / between the groups. There was also a significant increase in the amount of total proteins (79%) in the control group and polymorphonuclear cells (47%) in the ADSCs + PRP group. In view of the results, we observed that therapies with ADSCs + PRP and only ADSCs are safe and effective for improving pain, functional capacity and joint inflammation in patients with OA. However, the use of ADSCs + PRP is considered the most promising treatment option in helping to manage this pathology.
id UFMS_0bf16f18584ab0e5659d9d4ebcbc6d85
oai_identifier_str oai:repositorio.ufms.br:123456789/3707
network_acronym_str UFMS
network_name_str Repositório Institucional da UFMS
repository_id_str 2124
spelling 2021-05-24T11:42:12Z2021-09-30T19:55:51Z2021https://repositorio.ufms.br/handle/123456789/3707The increase in individuals with Osteoarthritis (OA) has generated an increase in public spending on monitoring and maintaining treatments, which have lasted for years and are still not resolvable. This study is the first cell therapy proposal for OA carried out in the Brazilian Public Health System, in the State of Mato Grosso do Sul. The objective of the work was divided into two stages: the laboratory which aimed to analyze the quality of adipose-derived stem cells (ADSCs ) cultured from the 1st to the 10th passage, and later the clinic, which in an unprecedented way analyzed and compared 4 types of interventions in patients with OA: platelet-rich plasma (PRP), ADSCs, ADSCs + PRP and the standard surgical procedure performed in our hospital. The analysis of the ADSCs passages was performed by immunophenotyping assay, cell differentiation, comet assay, cytological and molecular cell death assay, morphological analysis, apoptosis, membrane integrity and viability in the flow cytometer. The evaluation of the patients was carried out by applying the questionnaires of WOMAC, SF-36 and EVA, and by analyzing the synovial fluid (inflammatory cytokines, enzymatic, colorimetric and viscosity analysis). Regarding the quality of ADSCs, the trypsinization process carried out during cultivation did not interfere with the genomic stability of the cells. But as of the 6th pass, ADSCs started to modify its capacity for juxtaposed deposition and proliferation in monolayer, as well as increasing its size. Cell viability was significantly reduced only on the 10th passage, and it was proven that this was due to apoptosis due to changes in membrane permeability. The results of the questionnaires applied to the patients, showed a greater improvement in the scores of the analyzed domains (initial x final evaluation / by group and final values / between groups) in the ADSCs + PRP group, followed by the ADSCs and PRP group. In the evaluation of inflammatory cytokines, there was a significant reduction (initial x final / per group) of the IL-1b cytokine only in the ADSCs + PRP (40%) and ADSCs (31%) groups, of IL-6 in the ADSCs + PRP group ( 72%), IL-8 in the ADSCs + PRP (50%) and ADSCs (31%) groups, and TNF in the ADSCs + PRP group (46%). Only the reductions in the ADSCs + PRP group were considered significant in the analysis of the final values / between the groups. There was also a significant increase in the amount of total proteins (79%) in the control group and polymorphonuclear cells (47%) in the ADSCs + PRP group. In view of the results, we observed that therapies with ADSCs + PRP and only ADSCs are safe and effective for improving pain, functional capacity and joint inflammation in patients with OA. However, the use of ADSCs + PRP is considered the most promising treatment option in helping to manage this pathology.O aumento de indivíduos com Osteoartrite (OA) tem gerado aumento de gastos públicos com o acompanhamento e manutenção dos tratamentos, que perduraram anos e mesmo assim não são resolutivos. Este estudo é a primeira proposta de terapia celular para OA realizada no Sistema Público de Saúde Brasileiro, no Estado de Mato Grosso do Sul. O objetivo do trabalho foi dividido em duas etapas: a laboratorial que visou analisar a qualidade das células tronco mesenquimais (CTM) cultivadas da 1ª a 10ª passagem, e posteriormente a clínica, que de forma inédita analisou e comparou 4 tipos de intervenções em pacientes com OA: plasma rico em plaquetas (PRP), CTM, CTM+PRP e o procedimento cirúrgico padrão realizado em nosso hospital. A análise das passagens das CTM foi feita por ensaio de imunofenotipagem, diferenciação celular, ensaio de cometa, ensaio de morte celular citológico e molecular, análise morfológica, apoptose, integridade de membrana e viabilidade no citometro de fluxo. A avaliação dos pacientes foi realizada pela aplicação dos questionários WOMAC, SF-36 e EVA, e pela análise do líquido sinovial (citocinas inflamatórias, análise enzimática, colorimétrica e viscosidade). Em relação a qualidade das CTM, o processo de tripsinização realizado durante o cultivo não interferiu na estabilidade genômica das células. Mas a partir da 6ª passagem, as CTM começaram a modificar sua capacidade de deposição justaposta e proliferação em monocamada, assim como aumentaram seu tamanho. A viabilidade celular sofreu diminuição significativa apenas na 10º passagem, e comprovou-se que isso acontecia por apoptose devido a alteração da permeabilidade de membrana. Os resultados dos questionários aplicados nos pacientes, apresentaram uma maior melhora dos escores dos domínios analisados (avaliação inicial x final/por grupo e valores finais/ entre os grupos) no grupo CTM+PRP, seguido do grupo CTM e PRP. Na avaliação das citocinas inflamatórias, houve redução significativa (avaliação inicial x final/por grupo) da citocina IL-1b apenas nos grupo CTM+PRP (40%) e CTM (31%), da IL-6 no grupo CTM+PRP (72%), da IL-8 nos grupo CTM+PRP (50%) e CTM (31%), e TNF no grupo CTM+PRP (46%). Apenas as reduções do grupo CTM+PRP foram consideradas significativas na análise dos valores finais/ entre os grupos. Houve também aumento significativo da quantidade de proteinas totais (79%) no grupo controle e células polimorfonucleares (47%) do grupo CTM+PRP. Diante dos resultados, observamos que as terapias com CTM+PRP e apenas CTM são seguras e efetivas para a melhora do quadro álgico, da capacidade funcional e da inflamação articular em pacientes com OA. Entretanto, considera-se o uso de CTM+PRP como a alternativa mais promissora no auxílio do manejo desta patologia.Fundação Universidade Federal de Mato Grosso do SulUFMSBrasilTerapia CelularDor ArticularInflamaçãoOrtopediaMedicina Regenerativa.Efeitos da terapia celular, com células tronco mesenquimais do tecido adiposo, e do infiltrado de plasma rico em plaquetas no tratamento da osteoartrite não responsiva ao tratamento medicamentosoinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisRodrigo Juliano OliveiraLaynna de Carvalho Schweichinfo:eu-repo/semantics/openAccessporreponame:Repositório Institucional da UFMSinstname:Universidade Federal de Mato Grosso do Sul (UFMS)instacron:UFMSTHUMBNAILTese de doutorado Laynna Schweich-Adami versao final.pdf.jpgTese de doutorado Laynna Schweich-Adami versao final.pdf.jpgGenerated Thumbnailimage/jpeg1199https://repositorio.ufms.br/bitstream/123456789/3707/3/Tese%20de%20doutorado%20Laynna%20Schweich-Adami%20versao%20final.pdf.jpgc01fe1827c6301333e8f7b52a789bb3bMD53TEXTTese de doutorado Laynna Schweich-Adami versao final.pdf.txtTese de doutorado Laynna Schweich-Adami versao final.pdf.txtExtracted texttext/plain178902https://repositorio.ufms.br/bitstream/123456789/3707/2/Tese%20de%20doutorado%20Laynna%20Schweich-Adami%20versao%20final.pdf.txt99c9282aed2c966fa66ee14eeae080d9MD52ORIGINALTese de doutorado Laynna Schweich-Adami versao final.pdfTese de doutorado Laynna Schweich-Adami versao final.pdfapplication/pdf7832998https://repositorio.ufms.br/bitstream/123456789/3707/1/Tese%20de%20doutorado%20Laynna%20Schweich-Adami%20versao%20final.pdf89cefef84c62cbf361c5b81c820dee5aMD51123456789/37072021-09-30 15:55:51.588oai:repositorio.ufms.br:123456789/3707Repositório InstitucionalPUBhttps://repositorio.ufms.br/oai/requestri.prograd@ufms.bropendoar:21242021-09-30T19:55:51Repositório Institucional da UFMS - Universidade Federal de Mato Grosso do Sul (UFMS)false
dc.title.pt_BR.fl_str_mv Efeitos da terapia celular, com células tronco mesenquimais do tecido adiposo, e do infiltrado de plasma rico em plaquetas no tratamento da osteoartrite não responsiva ao tratamento medicamentoso
title Efeitos da terapia celular, com células tronco mesenquimais do tecido adiposo, e do infiltrado de plasma rico em plaquetas no tratamento da osteoartrite não responsiva ao tratamento medicamentoso
spellingShingle Efeitos da terapia celular, com células tronco mesenquimais do tecido adiposo, e do infiltrado de plasma rico em plaquetas no tratamento da osteoartrite não responsiva ao tratamento medicamentoso
Laynna de Carvalho Schweich
Terapia Celular
Dor Articular
Inflamação
Ortopedia
Medicina Regenerativa.
title_short Efeitos da terapia celular, com células tronco mesenquimais do tecido adiposo, e do infiltrado de plasma rico em plaquetas no tratamento da osteoartrite não responsiva ao tratamento medicamentoso
title_full Efeitos da terapia celular, com células tronco mesenquimais do tecido adiposo, e do infiltrado de plasma rico em plaquetas no tratamento da osteoartrite não responsiva ao tratamento medicamentoso
title_fullStr Efeitos da terapia celular, com células tronco mesenquimais do tecido adiposo, e do infiltrado de plasma rico em plaquetas no tratamento da osteoartrite não responsiva ao tratamento medicamentoso
title_full_unstemmed Efeitos da terapia celular, com células tronco mesenquimais do tecido adiposo, e do infiltrado de plasma rico em plaquetas no tratamento da osteoartrite não responsiva ao tratamento medicamentoso
title_sort Efeitos da terapia celular, com células tronco mesenquimais do tecido adiposo, e do infiltrado de plasma rico em plaquetas no tratamento da osteoartrite não responsiva ao tratamento medicamentoso
author Laynna de Carvalho Schweich
author_facet Laynna de Carvalho Schweich
author_role author
dc.contributor.advisor1.fl_str_mv Rodrigo Juliano Oliveira
dc.contributor.author.fl_str_mv Laynna de Carvalho Schweich
contributor_str_mv Rodrigo Juliano Oliveira
dc.subject.por.fl_str_mv Terapia Celular
Dor Articular
Inflamação
Ortopedia
Medicina Regenerativa.
topic Terapia Celular
Dor Articular
Inflamação
Ortopedia
Medicina Regenerativa.
description The increase in individuals with Osteoarthritis (OA) has generated an increase in public spending on monitoring and maintaining treatments, which have lasted for years and are still not resolvable. This study is the first cell therapy proposal for OA carried out in the Brazilian Public Health System, in the State of Mato Grosso do Sul. The objective of the work was divided into two stages: the laboratory which aimed to analyze the quality of adipose-derived stem cells (ADSCs ) cultured from the 1st to the 10th passage, and later the clinic, which in an unprecedented way analyzed and compared 4 types of interventions in patients with OA: platelet-rich plasma (PRP), ADSCs, ADSCs + PRP and the standard surgical procedure performed in our hospital. The analysis of the ADSCs passages was performed by immunophenotyping assay, cell differentiation, comet assay, cytological and molecular cell death assay, morphological analysis, apoptosis, membrane integrity and viability in the flow cytometer. The evaluation of the patients was carried out by applying the questionnaires of WOMAC, SF-36 and EVA, and by analyzing the synovial fluid (inflammatory cytokines, enzymatic, colorimetric and viscosity analysis). Regarding the quality of ADSCs, the trypsinization process carried out during cultivation did not interfere with the genomic stability of the cells. But as of the 6th pass, ADSCs started to modify its capacity for juxtaposed deposition and proliferation in monolayer, as well as increasing its size. Cell viability was significantly reduced only on the 10th passage, and it was proven that this was due to apoptosis due to changes in membrane permeability. The results of the questionnaires applied to the patients, showed a greater improvement in the scores of the analyzed domains (initial x final evaluation / by group and final values / between groups) in the ADSCs + PRP group, followed by the ADSCs and PRP group. In the evaluation of inflammatory cytokines, there was a significant reduction (initial x final / per group) of the IL-1b cytokine only in the ADSCs + PRP (40%) and ADSCs (31%) groups, of IL-6 in the ADSCs + PRP group ( 72%), IL-8 in the ADSCs + PRP (50%) and ADSCs (31%) groups, and TNF in the ADSCs + PRP group (46%). Only the reductions in the ADSCs + PRP group were considered significant in the analysis of the final values / between the groups. There was also a significant increase in the amount of total proteins (79%) in the control group and polymorphonuclear cells (47%) in the ADSCs + PRP group. In view of the results, we observed that therapies with ADSCs + PRP and only ADSCs are safe and effective for improving pain, functional capacity and joint inflammation in patients with OA. However, the use of ADSCs + PRP is considered the most promising treatment option in helping to manage this pathology.
publishDate 2021
dc.date.accessioned.fl_str_mv 2021-05-24T11:42:12Z
dc.date.available.fl_str_mv 2021-09-30T19:55:51Z
dc.date.issued.fl_str_mv 2021
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 https://repositorio.ufms.br/handle/123456789/3707
url https://repositorio.ufms.br/handle/123456789/3707
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Fundação Universidade Federal de Mato Grosso do Sul
dc.publisher.initials.fl_str_mv UFMS
dc.publisher.country.fl_str_mv Brasil
publisher.none.fl_str_mv Fundação Universidade Federal de Mato Grosso do Sul
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFMS
instname:Universidade Federal de Mato Grosso do Sul (UFMS)
instacron:UFMS
instname_str Universidade Federal de Mato Grosso do Sul (UFMS)
instacron_str UFMS
institution UFMS
reponame_str Repositório Institucional da UFMS
collection Repositório Institucional da UFMS
bitstream.url.fl_str_mv https://repositorio.ufms.br/bitstream/123456789/3707/3/Tese%20de%20doutorado%20Laynna%20Schweich-Adami%20versao%20final.pdf.jpg
https://repositorio.ufms.br/bitstream/123456789/3707/2/Tese%20de%20doutorado%20Laynna%20Schweich-Adami%20versao%20final.pdf.txt
https://repositorio.ufms.br/bitstream/123456789/3707/1/Tese%20de%20doutorado%20Laynna%20Schweich-Adami%20versao%20final.pdf
bitstream.checksum.fl_str_mv c01fe1827c6301333e8f7b52a789bb3b
99c9282aed2c966fa66ee14eeae080d9
89cefef84c62cbf361c5b81c820dee5a
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
MD5
repository.name.fl_str_mv Repositório Institucional da UFMS - Universidade Federal de Mato Grosso do Sul (UFMS)
repository.mail.fl_str_mv ri.prograd@ufms.br
_version_ 1797952992344801280