Efeito de um formulado desenvolvido com guaraná e semente de açaí em modelos experimentais pós-cirúrgicos: estudo da reparação in vitro e in vivo

Detalhes bibliográficos
Autor(a) principal: Felin, Fellipe Danezi
Data de Publicação: 2021
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
dARK ID: ark:/26339/0013000016v2t
Texto Completo: http://repositorio.ufsm.br/handle/1/23544
Resumo: Introduction: Tissue repair is a complex process that requires activation of mediators and mechanisms related to healing and regeneration. Healing involves inflammation, fibroblast proliferation and collagen remodeling. Studies have elucidated the modulation of repair at the molecular and biochemical level in an attempt to avoid pathological scarring. The repair can be influenced by the patient's nutritional status and, therefore, be stimulated by products originating from plants, including guarana and açaí, which have a healing effect by modulating the inflammatory and proliferative phases. It is possible that the combination of these results in a potential innovative pharmacological product, with high healing power. Objective: To evaluate the effect of the formula, developed with guarana and açaí seeds, in an experimental model of post-surgical repair, in vitro and in vivo. Methods: The study does not need to be approved by the Ethics Committee. Initially, the conjugate extract Guaraná-Açai (GA) was developed using hot water and citric acid, the extract was obtained and the chemical analysis was carried out through mass spectrophotometry. The antioxidant and genoprotective capacity of GA were analyzed respectively by the DPPH and Gemo tests. A pharmacological concentration-response curve (1, 3, 5, 10 and 30 μg / mL) in fibroblasts (HFF-1) and keratinocytes (HaCAT) both commercially obtained was performed and the concentration of choice for the other tests was 5 μg / mL. In fibroblasts, the strach assay was performed, the so-called in vitro wound assay, after 3,6, 24 and 72 hours of injury induction and treatment, oxidative markers were analyzed, apoptotic marker after 24 hours, total collagen after 24, 48 and 72 hours and analysis of the expression of genes related to proliferation, collagen and inflammation after 24 hours. A second in vivo protocol was performed using Eisenia fetida earthworm after 1,3,6,12 and 24 hours of the surgical caudal incision between the third and fifth posterior segment, and addition of treatment with GA or phosphate buffer for the control animals, macroscopic and histological analysis was performed using Masson staining -Goldner.The SOX-4 gene was measured via RT-PCR after 3 and 24 hours. Results: Sixteen bioactive molecules, including some substances never previously described in the isolated extracts, were identified. All concentrations tested in the cell model exhibited non-cytotoxic and antioxidant effects in addition to genoprotective. The concentration of choice 5 μg / mL showed pro-regeneration effects in an in vitro model. The GA extract accelerated the healing processes observed in cells and through macroscopic and histological analysis in earthworms. Conclusion: Despite the methodological limitations, these results indicated that the GA extract has the potential to accelerate the healing/regeneration of wounds induced in vitro or surgically in vivo. More studies need to be carried out so that these results can become a product to be made available to the population, bringing benefits to society as a whole.
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spelling Efeito de um formulado desenvolvido com guaraná e semente de açaí em modelos experimentais pós-cirúrgicos: estudo da reparação in vitro e in vivoEffect of a formula developed with guarana and açaí seed in post-surgical experimental models: in vitro and in vivo repair studyFerida operatóriaCicatrizaçãoColagenizaçãoRegeneraçãoCirurgiaReparoOperative woundHealingCollagenizationRegenerationSurgeryRepairCNPQ::CIENCIAS DA SAUDEIntroduction: Tissue repair is a complex process that requires activation of mediators and mechanisms related to healing and regeneration. Healing involves inflammation, fibroblast proliferation and collagen remodeling. Studies have elucidated the modulation of repair at the molecular and biochemical level in an attempt to avoid pathological scarring. The repair can be influenced by the patient's nutritional status and, therefore, be stimulated by products originating from plants, including guarana and açaí, which have a healing effect by modulating the inflammatory and proliferative phases. It is possible that the combination of these results in a potential innovative pharmacological product, with high healing power. Objective: To evaluate the effect of the formula, developed with guarana and açaí seeds, in an experimental model of post-surgical repair, in vitro and in vivo. Methods: The study does not need to be approved by the Ethics Committee. Initially, the conjugate extract Guaraná-Açai (GA) was developed using hot water and citric acid, the extract was obtained and the chemical analysis was carried out through mass spectrophotometry. The antioxidant and genoprotective capacity of GA were analyzed respectively by the DPPH and Gemo tests. A pharmacological concentration-response curve (1, 3, 5, 10 and 30 μg / mL) in fibroblasts (HFF-1) and keratinocytes (HaCAT) both commercially obtained was performed and the concentration of choice for the other tests was 5 μg / mL. In fibroblasts, the strach assay was performed, the so-called in vitro wound assay, after 3,6, 24 and 72 hours of injury induction and treatment, oxidative markers were analyzed, apoptotic marker after 24 hours, total collagen after 24, 48 and 72 hours and analysis of the expression of genes related to proliferation, collagen and inflammation after 24 hours. A second in vivo protocol was performed using Eisenia fetida earthworm after 1,3,6,12 and 24 hours of the surgical caudal incision between the third and fifth posterior segment, and addition of treatment with GA or phosphate buffer for the control animals, macroscopic and histological analysis was performed using Masson staining -Goldner.The SOX-4 gene was measured via RT-PCR after 3 and 24 hours. Results: Sixteen bioactive molecules, including some substances never previously described in the isolated extracts, were identified. All concentrations tested in the cell model exhibited non-cytotoxic and antioxidant effects in addition to genoprotective. The concentration of choice 5 μg / mL showed pro-regeneration effects in an in vitro model. The GA extract accelerated the healing processes observed in cells and through macroscopic and histological analysis in earthworms. Conclusion: Despite the methodological limitations, these results indicated that the GA extract has the potential to accelerate the healing/regeneration of wounds induced in vitro or surgically in vivo. More studies need to be carried out so that these results can become a product to be made available to the population, bringing benefits to society as a whole.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESConselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqFundação de Amparo à Pesquisa do Estado do Amazonas - FapeamIntrodução: O reparo dos tecidos é um processo complexo que exige ativação de mediadores e de mecanismos relacionados à cicatrização e à regeneração. A cicatrização envolve inflamação, proliferação de fibroblastos e remodelagem do colágeno. Estudos têm elucidado a modulação do reparo a nível molecular e bioquímico na tentativa de evitar cicatrizes patológicas. O reparo pode ser influenciado pelo estado nutricional do paciente e por isso ser estimulado por produtos originários de plantas, incluindo o guaraná e o açaí, que possuem efeito cicatrizante por modularem as fases inflamatória e proliferativa. É possível que a combinação destes, resulte em um potencial produto farmacológico inovador, com elevado poder cicatricial. Objetivo: Avaliar o efeito de um formulado, desenvolvido com semente de guaraná e de açaí, em modelo experimental de reparo pós-cirúrgico, in vitro e in vivo. Métodos: O estudo não necessita de aprovação em Comitê de ética. Inicialmente foi desenvolvido o extrato conjugado Guaraná- Açai (GA) com uso de água quente e ácido cítrico, o extrato foi obtido e a análise química foi feita através de espectrofotometria de massa. A capacidade antioxidante e genoprotetora do GA foi analisada respectivamente pelo teste DPPH e Gemo. Uma curva farmacológica concentração-resposta (1, 3, 5, 10 e 30 μg / mL) em fibroblastos (HFF-1) e queratinócitos (HaCAT) ambos obtidos comercialmente foi realizada e a concentração de escolha para os demais testes foi 5 μg / mL. Em fibroblastos então realizou-se o strach assay, o chamado ensaio de ferida in vitro, após 3,6,24 e 72 horas da indução da lesão e tratamento, foram analisados marcadores oxidativos, marcador apoptótico após 24 horas, colágeno total após 24, 48 e 72 horas e a análise da expressão de genes relacionados a proliferação, colágeno e inflamação após 24 horas. Um segundo protocolo in vivo foi realizado utilizando minhoca Eisenia fetida após 1,3,6,12 e 24 horas da incisão caudal cirúrgica entre o terceiro e quinto segmento posterior e adição do tratamento com GA ou tampão fosfato para os animais controle foi realizada a análise macroscópica e histológica utilizando a coloração Masson-Goldner.O gene SOX-4 foi mensurado via RT-PCR após 3 e 24 horas. Resultados: Dezesseis moléculas bioativas, incluindo algumas substâncias nunca descritas anteriormente nos extratos isolados foram identificadas. Todas as concentrações testadas no modelo celular exibiram efeitos não citotoxicos e antioxidantes além de genoprotetores. A concetração de escolha 5 μg / mL apresentou efeitos pró-regeneração em modelo in vitro. O extrato de GA acelerou os processos de cicatrização observados em células e por meio de análises macroscópicas e histológicas em minhocas. Conclusão: Apesar das limitações metodológicas, estes resultados indicaram que o extrato de GA tem potencial acelerador da cicatrização/ regeneração de feridas induzidas in vitro ou cirurgicamente in vivo. Mais estudos precisam ser realizados para que este resultados possam se transformar em um produto a ser disponibilizado a população, trazendo benefícios a sociedade como um todo.Universidade Federal de Santa MariaBrasilCiências da SaúdeUFSMPrograma de Pós-Graduação em Ciências da SaúdeCentro de Ciências da SaúdeRibeiro, Tiango Aguiarhttp://lattes.cnpq.br/5738745231283624Cruz, Ivana Beatrice Mânica Dahttp://lattes.cnpq.br/3426369324110716Pellenz, Neida Luiza KasparyPiccoli, Jacqueline da Costa EscobarFelin, Fellipe Danezi2022-01-17T12:23:13Z2022-01-17T12:23:13Z2021-09-02info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/23544ark:/26339/0013000016v2tporAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2022-01-17T12:23:13Zoai:repositorio.ufsm.br:1/23544Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2022-01-17T12:23:13Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Efeito de um formulado desenvolvido com guaraná e semente de açaí em modelos experimentais pós-cirúrgicos: estudo da reparação in vitro e in vivo
Effect of a formula developed with guarana and açaí seed in post-surgical experimental models: in vitro and in vivo repair study
title Efeito de um formulado desenvolvido com guaraná e semente de açaí em modelos experimentais pós-cirúrgicos: estudo da reparação in vitro e in vivo
spellingShingle Efeito de um formulado desenvolvido com guaraná e semente de açaí em modelos experimentais pós-cirúrgicos: estudo da reparação in vitro e in vivo
Felin, Fellipe Danezi
Ferida operatória
Cicatrização
Colagenização
Regeneração
Cirurgia
Reparo
Operative wound
Healing
Collagenization
Regeneration
Surgery
Repair
CNPQ::CIENCIAS DA SAUDE
title_short Efeito de um formulado desenvolvido com guaraná e semente de açaí em modelos experimentais pós-cirúrgicos: estudo da reparação in vitro e in vivo
title_full Efeito de um formulado desenvolvido com guaraná e semente de açaí em modelos experimentais pós-cirúrgicos: estudo da reparação in vitro e in vivo
title_fullStr Efeito de um formulado desenvolvido com guaraná e semente de açaí em modelos experimentais pós-cirúrgicos: estudo da reparação in vitro e in vivo
title_full_unstemmed Efeito de um formulado desenvolvido com guaraná e semente de açaí em modelos experimentais pós-cirúrgicos: estudo da reparação in vitro e in vivo
title_sort Efeito de um formulado desenvolvido com guaraná e semente de açaí em modelos experimentais pós-cirúrgicos: estudo da reparação in vitro e in vivo
author Felin, Fellipe Danezi
author_facet Felin, Fellipe Danezi
author_role author
dc.contributor.none.fl_str_mv Ribeiro, Tiango Aguiar
http://lattes.cnpq.br/5738745231283624
Cruz, Ivana Beatrice Mânica Da
http://lattes.cnpq.br/3426369324110716
Pellenz, Neida Luiza Kaspary
Piccoli, Jacqueline da Costa Escobar
dc.contributor.author.fl_str_mv Felin, Fellipe Danezi
dc.subject.por.fl_str_mv Ferida operatória
Cicatrização
Colagenização
Regeneração
Cirurgia
Reparo
Operative wound
Healing
Collagenization
Regeneration
Surgery
Repair
CNPQ::CIENCIAS DA SAUDE
topic Ferida operatória
Cicatrização
Colagenização
Regeneração
Cirurgia
Reparo
Operative wound
Healing
Collagenization
Regeneration
Surgery
Repair
CNPQ::CIENCIAS DA SAUDE
description Introduction: Tissue repair is a complex process that requires activation of mediators and mechanisms related to healing and regeneration. Healing involves inflammation, fibroblast proliferation and collagen remodeling. Studies have elucidated the modulation of repair at the molecular and biochemical level in an attempt to avoid pathological scarring. The repair can be influenced by the patient's nutritional status and, therefore, be stimulated by products originating from plants, including guarana and açaí, which have a healing effect by modulating the inflammatory and proliferative phases. It is possible that the combination of these results in a potential innovative pharmacological product, with high healing power. Objective: To evaluate the effect of the formula, developed with guarana and açaí seeds, in an experimental model of post-surgical repair, in vitro and in vivo. Methods: The study does not need to be approved by the Ethics Committee. Initially, the conjugate extract Guaraná-Açai (GA) was developed using hot water and citric acid, the extract was obtained and the chemical analysis was carried out through mass spectrophotometry. The antioxidant and genoprotective capacity of GA were analyzed respectively by the DPPH and Gemo tests. A pharmacological concentration-response curve (1, 3, 5, 10 and 30 μg / mL) in fibroblasts (HFF-1) and keratinocytes (HaCAT) both commercially obtained was performed and the concentration of choice for the other tests was 5 μg / mL. In fibroblasts, the strach assay was performed, the so-called in vitro wound assay, after 3,6, 24 and 72 hours of injury induction and treatment, oxidative markers were analyzed, apoptotic marker after 24 hours, total collagen after 24, 48 and 72 hours and analysis of the expression of genes related to proliferation, collagen and inflammation after 24 hours. A second in vivo protocol was performed using Eisenia fetida earthworm after 1,3,6,12 and 24 hours of the surgical caudal incision between the third and fifth posterior segment, and addition of treatment with GA or phosphate buffer for the control animals, macroscopic and histological analysis was performed using Masson staining -Goldner.The SOX-4 gene was measured via RT-PCR after 3 and 24 hours. Results: Sixteen bioactive molecules, including some substances never previously described in the isolated extracts, were identified. All concentrations tested in the cell model exhibited non-cytotoxic and antioxidant effects in addition to genoprotective. The concentration of choice 5 μg / mL showed pro-regeneration effects in an in vitro model. The GA extract accelerated the healing processes observed in cells and through macroscopic and histological analysis in earthworms. Conclusion: Despite the methodological limitations, these results indicated that the GA extract has the potential to accelerate the healing/regeneration of wounds induced in vitro or surgically in vivo. More studies need to be carried out so that these results can become a product to be made available to the population, bringing benefits to society as a whole.
publishDate 2021
dc.date.none.fl_str_mv 2021-09-02
2022-01-17T12:23:13Z
2022-01-17T12:23:13Z
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 http://repositorio.ufsm.br/handle/1/23544
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url http://repositorio.ufsm.br/handle/1/23544
identifier_str_mv ark:/26339/0013000016v2t
dc.language.iso.fl_str_mv por
language por
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.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Ciências da Saúde
UFSM
Programa de Pós-Graduação em Ciências da Saúde
Centro de Ciências da Saúde
publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Ciências da Saúde
UFSM
Programa de Pós-Graduação em Ciências da Saúde
Centro de Ciências da Saúde
dc.source.none.fl_str_mv reponame:Manancial - Repositório Digital da UFSM
instname:Universidade Federal de Santa Maria (UFSM)
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instname_str Universidade Federal de Santa Maria (UFSM)
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institution UFSM
reponame_str Manancial - Repositório Digital da UFSM
collection Manancial - Repositório Digital da UFSM
repository.name.fl_str_mv Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)
repository.mail.fl_str_mv atendimento.sib@ufsm.br||tedebc@gmail.com
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