Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativo

Detalhes bibliográficos
Autor(a) principal: Moreira Filho, Raimundo Nonato Fernandes
Data de Publicação: 2021
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
Texto Completo: http://www.repositorio.ufc.br/handle/riufc/62413
Resumo: Natural-based dressings are of considerable interest in the pharmaceutical industry because they have low body rejection. Alginate, a polysaccharide found in the cell walls of brown algae, has excellent properties for use as biomaterial, as it is biodegradable, biocompatible and non-toxic. Alginate dressings are already a therapeutic reality, however they act only by maintaining the proper moisture of the place. In order to improve its therapeutic properties, it was proposed the incorporation of an enzyme (papain) to accelerate and improve wound healing. Papain may be found in the composition of ointments or creams for the treatment of wounds, as it is capable of promoting the debridement of the devitalized or necrotic tissues near the lesion, accelerating the healing process. The development of a dressing based on alginate and papain adds the healing properties of both materials that will potentially act to improve the healing process. The adsorption of the enzyme on a support can stabilize its structure and allow a controlled release. In the present work, papain was immobilized by physical adsorption and the best conditions for immobilization (initial concentration, temperature and pH) were evaluated and the yield of immobilization was measured by the Bradford test. It was analyzed whether the enzyme remained active after immobilization as well as measurements were taken over time to assess its stability. Desorption of the protein and its release using Franz cell was studied. Characterization tests (FTIR and SEM) and in vitro biological assays were performed (hemolysis, cytotoxicity and antibacterial). It was found that the best immobilization conditions were in neutral medium, 20 mg/mL and 25 ° C. The enzyme remained active after immobilization (5.89 U/mg) and over time it was observed that immobilized papain had a smaller decrease in relative activity compared to solution (29.15% of its initial activity versus 6.31% over 28 days). Desorption showed that 97% of the enzyme was released after 6 hours; in the Franz cell, 64.1% of the enzyme was released after 24 hours. The biological assays point to a material with low cytotoxicity and not hemolytic. Based on these initial tests, it was possible to obtain a biomaterial with good potential to be applied as a bioactive dressing.
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spelling Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativoCharacterization and evaluation of calcium alginate membranes with immobilized papain for use as a bioactive dressingCurativoAlginatoPapaínaAdsorçãoNatural-based dressings are of considerable interest in the pharmaceutical industry because they have low body rejection. Alginate, a polysaccharide found in the cell walls of brown algae, has excellent properties for use as biomaterial, as it is biodegradable, biocompatible and non-toxic. Alginate dressings are already a therapeutic reality, however they act only by maintaining the proper moisture of the place. In order to improve its therapeutic properties, it was proposed the incorporation of an enzyme (papain) to accelerate and improve wound healing. Papain may be found in the composition of ointments or creams for the treatment of wounds, as it is capable of promoting the debridement of the devitalized or necrotic tissues near the lesion, accelerating the healing process. The development of a dressing based on alginate and papain adds the healing properties of both materials that will potentially act to improve the healing process. The adsorption of the enzyme on a support can stabilize its structure and allow a controlled release. In the present work, papain was immobilized by physical adsorption and the best conditions for immobilization (initial concentration, temperature and pH) were evaluated and the yield of immobilization was measured by the Bradford test. It was analyzed whether the enzyme remained active after immobilization as well as measurements were taken over time to assess its stability. Desorption of the protein and its release using Franz cell was studied. Characterization tests (FTIR and SEM) and in vitro biological assays were performed (hemolysis, cytotoxicity and antibacterial). It was found that the best immobilization conditions were in neutral medium, 20 mg/mL and 25 ° C. The enzyme remained active after immobilization (5.89 U/mg) and over time it was observed that immobilized papain had a smaller decrease in relative activity compared to solution (29.15% of its initial activity versus 6.31% over 28 days). Desorption showed that 97% of the enzyme was released after 6 hours; in the Franz cell, 64.1% of the enzyme was released after 24 hours. The biological assays point to a material with low cytotoxicity and not hemolytic. Based on these initial tests, it was possible to obtain a biomaterial with good potential to be applied as a bioactive dressing.FUNCAPCurativos a base de produtos naturais são de bastante interesse na indústria farmacêutica por apresentarem baixa rejeição pelo organismo. O alginato, um polissacarídeo encontrado nas paredes celulares de algas marrons, tem excelentes propriedades para o uso como biomaterial, devido a sua biodegradabilidade e biocompatibilidade. Curativos de alginato já são uma realidade terapêutica, porém, eles atuam apenas mantendo o ambiente adequado para a cicatrização. Com o intuito de melhorar suas propriedades, foi proposta a incorporação da papaína para acelerar e melhorar a cicatrização de feridas. Essa enzima já é usada na composição de pomadas e cremes para tratamento de feridas, pois é capaz de promover o desbridamento dos tecidos desvitalizados ou necróticos próximos à lesão, acelerando o processo de cicatrização. O desenvolvimento de um curativo a base de alginato e papaína agrega as propriedades cicatrizantes de ambos os materiais que atuarão melhorando o processo de cicatrização. No presente trabalho, a papaína foi imobilizada por adsorção física e foram avaliadas as melhores condições para imobilização (concentração inicial, temperatura e pH) e medido o rendimento da imobilização ao quantificar a proteína retida através do teste de Bradford. Foi feito um estudo da estabilidade enzimática após imobilização e ao longo do período de armazenamento (28 dias). A liberação da enzima foi quantificada utilizando célula de Franz. Foram feitos testes de caracterização química e morfológica (FTIR e MEV) e ensaios biológicos in vitro (hemólise, citotoxicidade e antibacteriano). Verificou-se que as melhores condições de imobilização foram encontradas em meio neutro, concentração inicial de 20 mg/mL de papaína e a 25 ⁰C. A enzima continuou ativa após imobilização (5,89 U/mg). Ao longo do tempo notou-se que a papaína imobilizada tem menor redução da atividade relativa comparada à em solução (manteve 29,15% da atividade inicial contra 6,31%). Foi constatado que 64,1% da enzima total imobilizada foi liberada após 24 horas. Os ensaios biológicos apontam para um material bioativo e não hemolítico. Com base nesses testes iniciais, foi possível obter um biomaterial com bom potencial para ser aplicado como curativo bioativo.Rosa, Morysleide de FreitasFeitosa, Judith Pessoa de AndradeMoreira Filho, Raimundo Nonato Fernandes2021-11-23T19:40:44Z2021-11-23T19:40:44Z2021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfMOREIRA FILHO, Raimundo Nonato Fernandes. Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativo. 2019. 62 f. Dissertação (Mestrado em Química) - Universidade Federal do Ceará, Fortaleza, 2019.http://www.repositorio.ufc.br/handle/riufc/62413porreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2021-11-23T19:40:44Zoai:repositorio.ufc.br:riufc/62413Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:27:47.659887Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.none.fl_str_mv Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativo
Characterization and evaluation of calcium alginate membranes with immobilized papain for use as a bioactive dressing
title Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativo
spellingShingle Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativo
Moreira Filho, Raimundo Nonato Fernandes
Curativo
Alginato
Papaína
Adsorção
title_short Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativo
title_full Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativo
title_fullStr Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativo
title_full_unstemmed Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativo
title_sort Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativo
author Moreira Filho, Raimundo Nonato Fernandes
author_facet Moreira Filho, Raimundo Nonato Fernandes
author_role author
dc.contributor.none.fl_str_mv Rosa, Morysleide de Freitas
Feitosa, Judith Pessoa de Andrade
dc.contributor.author.fl_str_mv Moreira Filho, Raimundo Nonato Fernandes
dc.subject.por.fl_str_mv Curativo
Alginato
Papaína
Adsorção
topic Curativo
Alginato
Papaína
Adsorção
description Natural-based dressings are of considerable interest in the pharmaceutical industry because they have low body rejection. Alginate, a polysaccharide found in the cell walls of brown algae, has excellent properties for use as biomaterial, as it is biodegradable, biocompatible and non-toxic. Alginate dressings are already a therapeutic reality, however they act only by maintaining the proper moisture of the place. In order to improve its therapeutic properties, it was proposed the incorporation of an enzyme (papain) to accelerate and improve wound healing. Papain may be found in the composition of ointments or creams for the treatment of wounds, as it is capable of promoting the debridement of the devitalized or necrotic tissues near the lesion, accelerating the healing process. The development of a dressing based on alginate and papain adds the healing properties of both materials that will potentially act to improve the healing process. The adsorption of the enzyme on a support can stabilize its structure and allow a controlled release. In the present work, papain was immobilized by physical adsorption and the best conditions for immobilization (initial concentration, temperature and pH) were evaluated and the yield of immobilization was measured by the Bradford test. It was analyzed whether the enzyme remained active after immobilization as well as measurements were taken over time to assess its stability. Desorption of the protein and its release using Franz cell was studied. Characterization tests (FTIR and SEM) and in vitro biological assays were performed (hemolysis, cytotoxicity and antibacterial). It was found that the best immobilization conditions were in neutral medium, 20 mg/mL and 25 ° C. The enzyme remained active after immobilization (5.89 U/mg) and over time it was observed that immobilized papain had a smaller decrease in relative activity compared to solution (29.15% of its initial activity versus 6.31% over 28 days). Desorption showed that 97% of the enzyme was released after 6 hours; in the Franz cell, 64.1% of the enzyme was released after 24 hours. The biological assays point to a material with low cytotoxicity and not hemolytic. Based on these initial tests, it was possible to obtain a biomaterial with good potential to be applied as a bioactive dressing.
publishDate 2021
dc.date.none.fl_str_mv 2021-11-23T19:40:44Z
2021-11-23T19:40:44Z
2021
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
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status_str publishedVersion
dc.identifier.uri.fl_str_mv MOREIRA FILHO, Raimundo Nonato Fernandes. Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativo. 2019. 62 f. Dissertação (Mestrado em Química) - Universidade Federal do Ceará, Fortaleza, 2019.
http://www.repositorio.ufc.br/handle/riufc/62413
identifier_str_mv MOREIRA FILHO, Raimundo Nonato Fernandes. Caracterização e avaliação de membranas de alginato de cálcio com papaína imobilizada para o uso como curativo bioativo. 2019. 62 f. Dissertação (Mestrado em Química) - Universidade Federal do Ceará, Fortaleza, 2019.
url http://www.repositorio.ufc.br/handle/riufc/62413
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instacron:UFC
instname_str Universidade Federal do Ceará (UFC)
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