Síntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastite

Detalhes bibliográficos
Ano de defesa: 2017
Autor(a) principal: Castro, Karine Cappuccio De lattes
Orientador(a): Campos, Maria Gabriela Nogueira lattes
Banca de defesa: Carvalho, Flávia Chiva, Marques, Rodrigo Fernando Costa
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Alfenas
Programa de Pós-Graduação: Programa de Pós-Graduação em Ciência e Engenharia de Materiais
Departamento: Instituto de Ciência e Tecnologia
País: Brasil
Palavras-chave em Português:
Área do conhecimento CNPq:
Link de acesso: https://repositorio.unifal-mg.edu.br/handle/123456789/1059
Resumo: Bovine mastitis causes a serious impact on the dairy sector, causing serious damage, both due to the functional impairment of the mammary gland, with reduced milk production. In addition, it causes serious risks to public health, as the major species of bacterial pathogens that cause mastitis are produce toxin. In this context, nanotechnology is a very interesting alternative to overcome the limitations of conventional mastitis treatments, through the creation of nanostructured devices that are able to direct the antimicrobial to the surface of the glandular epithelium and into the affected tissue in order to potentiate the effect of the antibiotic under bacteria. The use of chitosan and Polylactic Acid (PLA) for this type of application is due to its numerous properties, especially biocompatibility and biodegradability. Thus this work was aimed at obtaining and characterization of nanoparticles for controlled release of gentamicin (GEN) through the emulsification - solvent evaporation. The nanoparticles were characterized by Scanning Electron Microscopy (SEM) coupled to a Dispersive Energy Detector (EDS), Infrared Spectroscopy with Fourier Transform (FTIR), Dynamic Light Scattering (DLS) and Zeta Potential measurements. In addition, the efficiency of encapsulation of gentamicin, the in vitro release of gentamicin using the dialysis method and the antimicrobial activity through the disc diffusion test and the determination of the Minimum Inhibition Concentration (MIC) by the successive microdilution method, using Staphylococcus aureus, were evaluated. DLS analyses showed an average nanoparticle size between 150 and 400 nm, varying with the composition of the formulation. The SEM analyzes correlated well with the DLS measurements, by this analysis it was also possible to observe the formation of agglomerates under certain conditions of synthesis. By FTIR analysis through the characteristic bands and the enlargement and/or increase of the bands intensification, confirmed the interaction between the components of the formulation, suggesting the formation of a core-shell nanoparticle, where the PLA would be in the nucleus and the chitosan in the shell. The study of zeta potential as a function of pH showed that the particles have positive surface charges, which is attractive for cell adhesion. The encapsulation efficiency varied from 66 to 75%, the 1:0.25 sample was the most successful in the encapsulation, suggesting that gentamicin, although hydrophilic, was also bound to PLA (hydrophobic). ESD analysis confirmed the presence of sulfur in all formulations containing gentamicin sulfate, which also confirmed the encapsulation of the antibiotic. The release assays showed that the highest concentration of gentamicin was released in the first 6 hours and that in the period from 24 to 96 hours that concentration remained in equilibrium. Finally, the antimicrobial assay is performed with the suspensions used in vitro release test showed the growth inhibition of Staphylococcus aureus main cause of bovine mastitis, and the MIC obtained for all samples was consistent with the literature.
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spelling Castro, Karine Cappuccio Dehttp://lattes.cnpq.br/1741478379427600Carvalho, Flávia ChivaMarques, Rodrigo Fernando CostaCampos, Maria Gabriela Nogueirahttp://lattes.cnpq.br/01584321127108852018-01-05T19:59:39Z2017-07-27CASTRO, Karine Cappuccio de. Síntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastite. 2017. 63 f. Dissertação (Mestrado em Ciência e Engenharia de Materiais) - Universidade Federal de Alfenas, Poços de Caldas, MG, 2017.https://repositorio.unifal-mg.edu.br/handle/123456789/1059Bovine mastitis causes a serious impact on the dairy sector, causing serious damage, both due to the functional impairment of the mammary gland, with reduced milk production. In addition, it causes serious risks to public health, as the major species of bacterial pathogens that cause mastitis are produce toxin. In this context, nanotechnology is a very interesting alternative to overcome the limitations of conventional mastitis treatments, through the creation of nanostructured devices that are able to direct the antimicrobial to the surface of the glandular epithelium and into the affected tissue in order to potentiate the effect of the antibiotic under bacteria. The use of chitosan and Polylactic Acid (PLA) for this type of application is due to its numerous properties, especially biocompatibility and biodegradability. Thus this work was aimed at obtaining and characterization of nanoparticles for controlled release of gentamicin (GEN) through the emulsification - solvent evaporation. The nanoparticles were characterized by Scanning Electron Microscopy (SEM) coupled to a Dispersive Energy Detector (EDS), Infrared Spectroscopy with Fourier Transform (FTIR), Dynamic Light Scattering (DLS) and Zeta Potential measurements. In addition, the efficiency of encapsulation of gentamicin, the in vitro release of gentamicin using the dialysis method and the antimicrobial activity through the disc diffusion test and the determination of the Minimum Inhibition Concentration (MIC) by the successive microdilution method, using Staphylococcus aureus, were evaluated. DLS analyses showed an average nanoparticle size between 150 and 400 nm, varying with the composition of the formulation. The SEM analyzes correlated well with the DLS measurements, by this analysis it was also possible to observe the formation of agglomerates under certain conditions of synthesis. By FTIR analysis through the characteristic bands and the enlargement and/or increase of the bands intensification, confirmed the interaction between the components of the formulation, suggesting the formation of a core-shell nanoparticle, where the PLA would be in the nucleus and the chitosan in the shell. The study of zeta potential as a function of pH showed that the particles have positive surface charges, which is attractive for cell adhesion. The encapsulation efficiency varied from 66 to 75%, the 1:0.25 sample was the most successful in the encapsulation, suggesting that gentamicin, although hydrophilic, was also bound to PLA (hydrophobic). ESD analysis confirmed the presence of sulfur in all formulations containing gentamicin sulfate, which also confirmed the encapsulation of the antibiotic. The release assays showed that the highest concentration of gentamicin was released in the first 6 hours and that in the period from 24 to 96 hours that concentration remained in equilibrium. Finally, the antimicrobial assay is performed with the suspensions used in vitro release test showed the growth inhibition of Staphylococcus aureus main cause of bovine mastitis, and the MIC obtained for all samples was consistent with the literature.A mastite bovina ocasiona grave impacto no setor leiteiro, causando sérios prejuízos, tanto pelo comprometimento funcional da glândula mamária, como na redução da produção de leite. Além disso, causa sérios riscos à saúde pública, uma vez que as principais espécies de patógenos bacterianos causadores de mastite são produtoras de toxinas. Neste contexto, a nanotecnologia é uma alternativa bastante interessante para contornar as limitações dos tratamentos convencionais de mastite, através da criação de dispositivos nanoestruturados que sejam capazes de direcionar o antimicrobiano para a superfície do epitélio glandular e para o interior do tecido afetado, de forma a potencializar o efeito do antibiótico sob as bactérias. O uso da quitosana e do Ácido Polilático (PLA) para este tipo de aplicação se deve às suas inúmeras propriedades, em especial a biocompatibilidade e a biodegradabilidade. Desta forma, este trabalho teve como objetivo a obtenção e caracterização de diferentes formulações de nanopartículas de quitosana/PLA para liberação controlada de gentamicina (GEN), através da metodologia de emulsificação - evaporação de solvente. As nanopartículas foram caracterizadas por Microscopia Eletrônica de Varredura (MEV) acoplada a um Detector de Energia Dispersiva (EDS), Espectroscopia de Infravermelho com Transformada de Fourier (FTIR), Distribuição do Tamanho de Partículas (DLS) e Carga Superficial (Potencial Zeta). Ainda, foram avaliadas a eficiência de encapsulamento da gentamicina, a liberação de gentamicina in vitro pelo método de diálise e a atividade antimicrobiana através do teste de Difusão em Disco e da determinação da Concentração Mínima de Inibição (CMI) pelo método de microdiluição sucessiva, utilizando Staphylococcus aureus. As análises de DLS mostraram um tamanho médio de nanopartículas entre 150 e 400 nm, variando com a composição da formulação. As análises de MEV mostraram partículas esféricas de tamanho correspondente às medidas de DLS, por essa análise também foi possível observar a formação de aglomerados em determinadas condições de síntese. Pelas análises de FTIR, através das bandas características e do alargamento e/ou aumento da intensificação das bandas, foi possível confirmar a interação entre os componentes da formulação, sugerindo a formação de uma nanopartícula do tipo “core-shell”, onde o PLA estaria no núcleo e a quitosana na casca. Ainda, o estudo do potencial zeta em função do pH mostrou que as partículas possuem cargas de superfície positivas, provavelmente devido à presença de quitosana na casca, o que é atrativo para adesão celular. A eficiência de encapsulamento variou de 66 a 75%. A formulação 1:0,25 (w/w, PLA/quitosana) foi a que apresentou maior eficiência de encapsulamento, sugerindo que o PLA, apesar de hidrofóbico, influenciou na retenção da gentamicina (hidrofílica) na nanopartícula. A análise por EDS confirmou a presença de enxofre em todas as formulações contendo sulfato de gentamicina, o que também comprovou o encapsulamento do antibiótico. Os ensaios de liberação mostraram que a maior concentração de gentamicina foi liberada nas 6 primeiras horas de ensaio. Além disso, após 96 horas de ensaio, cerca de 90% da concentração inicial da gentamicina foi liberada. Por fim, o ensaio antimicrobiano realizado com as suspensões utilizadas o ensaio de liberação in vitro indicou a inibição do crescimento da Staphylococcus aureus, principal causadora da mastite bovina, bem como a CMI obtida para todas as amostras foi condizente com a literatura.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESapplication/pdfporUniversidade Federal de AlfenasPrograma de Pós-Graduação em Ciência e Engenharia de MateriaisUNIFAL-MGBrasilInstituto de Ciência e Tecnologiainfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-nd/4.0/Nanopartículas.Quitosana.Poli (Ácido Lático).Gentamicina.Mastite.ENGENHARIA DE MATERIAIS E METALURGICA::MATERIAIS NAO METALICOSSíntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastiteSynthesis of chitosan/PLA nanoparticles for controlled release of antibiotic in the treatment of mastitisinfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/publishedVersion-4297417259498638931600600600-52618770831920532972075167498588264571reponame:Repositório Institucional da Universidade Federal de Alfenas - RiUnifalinstname:Universidade Federal de Alfenas (UNIFAL)instacron:UNIFALCastro, Karine Cappuccio DeLICENSElicense.txtlicense.txttext/plain; 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dc.title.pt-BR.fl_str_mv Síntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastite
dc.title.alternative.eng.fl_str_mv Synthesis of chitosan/PLA nanoparticles for controlled release of antibiotic in the treatment of mastitis
title Síntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastite
spellingShingle Síntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastite
Castro, Karine Cappuccio De
Nanopartículas.
Quitosana.
Poli (Ácido Lático).
Gentamicina.
Mastite.
ENGENHARIA DE MATERIAIS E METALURGICA::MATERIAIS NAO METALICOS
title_short Síntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastite
title_full Síntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastite
title_fullStr Síntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastite
title_full_unstemmed Síntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastite
title_sort Síntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastite
author Castro, Karine Cappuccio De
author_facet Castro, Karine Cappuccio De
author_role author
dc.contributor.author.fl_str_mv Castro, Karine Cappuccio De
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/1741478379427600
dc.contributor.referee1.fl_str_mv Carvalho, Flávia Chiva
dc.contributor.referee2.fl_str_mv Marques, Rodrigo Fernando Costa
dc.contributor.advisor1.fl_str_mv Campos, Maria Gabriela Nogueira
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/0158432112710885
contributor_str_mv Carvalho, Flávia Chiva
Marques, Rodrigo Fernando Costa
Campos, Maria Gabriela Nogueira
dc.subject.por.fl_str_mv Nanopartículas.
Quitosana.
Poli (Ácido Lático).
Gentamicina.
Mastite.
topic Nanopartículas.
Quitosana.
Poli (Ácido Lático).
Gentamicina.
Mastite.
ENGENHARIA DE MATERIAIS E METALURGICA::MATERIAIS NAO METALICOS
dc.subject.cnpq.fl_str_mv ENGENHARIA DE MATERIAIS E METALURGICA::MATERIAIS NAO METALICOS
description Bovine mastitis causes a serious impact on the dairy sector, causing serious damage, both due to the functional impairment of the mammary gland, with reduced milk production. In addition, it causes serious risks to public health, as the major species of bacterial pathogens that cause mastitis are produce toxin. In this context, nanotechnology is a very interesting alternative to overcome the limitations of conventional mastitis treatments, through the creation of nanostructured devices that are able to direct the antimicrobial to the surface of the glandular epithelium and into the affected tissue in order to potentiate the effect of the antibiotic under bacteria. The use of chitosan and Polylactic Acid (PLA) for this type of application is due to its numerous properties, especially biocompatibility and biodegradability. Thus this work was aimed at obtaining and characterization of nanoparticles for controlled release of gentamicin (GEN) through the emulsification - solvent evaporation. The nanoparticles were characterized by Scanning Electron Microscopy (SEM) coupled to a Dispersive Energy Detector (EDS), Infrared Spectroscopy with Fourier Transform (FTIR), Dynamic Light Scattering (DLS) and Zeta Potential measurements. In addition, the efficiency of encapsulation of gentamicin, the in vitro release of gentamicin using the dialysis method and the antimicrobial activity through the disc diffusion test and the determination of the Minimum Inhibition Concentration (MIC) by the successive microdilution method, using Staphylococcus aureus, were evaluated. DLS analyses showed an average nanoparticle size between 150 and 400 nm, varying with the composition of the formulation. The SEM analyzes correlated well with the DLS measurements, by this analysis it was also possible to observe the formation of agglomerates under certain conditions of synthesis. By FTIR analysis through the characteristic bands and the enlargement and/or increase of the bands intensification, confirmed the interaction between the components of the formulation, suggesting the formation of a core-shell nanoparticle, where the PLA would be in the nucleus and the chitosan in the shell. The study of zeta potential as a function of pH showed that the particles have positive surface charges, which is attractive for cell adhesion. The encapsulation efficiency varied from 66 to 75%, the 1:0.25 sample was the most successful in the encapsulation, suggesting that gentamicin, although hydrophilic, was also bound to PLA (hydrophobic). ESD analysis confirmed the presence of sulfur in all formulations containing gentamicin sulfate, which also confirmed the encapsulation of the antibiotic. The release assays showed that the highest concentration of gentamicin was released in the first 6 hours and that in the period from 24 to 96 hours that concentration remained in equilibrium. Finally, the antimicrobial assay is performed with the suspensions used in vitro release test showed the growth inhibition of Staphylococcus aureus main cause of bovine mastitis, and the MIC obtained for all samples was consistent with the literature.
publishDate 2017
dc.date.issued.fl_str_mv 2017-07-27
dc.date.accessioned.fl_str_mv 2018-01-05T19:59:39Z
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format masterThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv CASTRO, Karine Cappuccio de. Síntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastite. 2017. 63 f. Dissertação (Mestrado em Ciência e Engenharia de Materiais) - Universidade Federal de Alfenas, Poços de Caldas, MG, 2017.
dc.identifier.uri.fl_str_mv https://repositorio.unifal-mg.edu.br/handle/123456789/1059
identifier_str_mv CASTRO, Karine Cappuccio de. Síntese de nanopartículas de quitosana/PLA para liberação controlada de antibiótico no tratamento de mastite. 2017. 63 f. Dissertação (Mestrado em Ciência e Engenharia de Materiais) - Universidade Federal de Alfenas, Poços de Caldas, MG, 2017.
url https://repositorio.unifal-mg.edu.br/handle/123456789/1059
dc.language.iso.fl_str_mv por
language por
dc.relation.department.fl_str_mv -4297417259498638931
dc.relation.confidence.fl_str_mv 600
600
600
dc.relation.cnpq.fl_str_mv -5261877083192053297
dc.relation.sponsorship.fl_str_mv 2075167498588264571
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Alfenas
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Ciência e Engenharia de Materiais
dc.publisher.initials.fl_str_mv UNIFAL-MG
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Instituto de Ciência e Tecnologia
publisher.none.fl_str_mv Universidade Federal de Alfenas
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal de Alfenas - RiUnifal
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