Síntese e caracterização de nanopartículas de quitosana carreando curcumina para avaliação de inativação fotodinâmica

Detalhes bibliográficos
Autor(a) principal: Laís Fernandes Aguilera
Data de Publicação: 2024
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFMS
Texto Completo: https://repositorio.ufms.br/handle/123456789/8965
Resumo: For a long time, humanity has faced several diseases caused by bacterial infections. The discovery of antibiotics completely changed modern medicine and extended human life expectancy. However, abuse and misuse of antibiotics over the years is one of the factors that leads to bacterial resistance reaching alarming levels. The development of a new generation of anti-infective medicines using natural antimicrobial products is very promising. Another promising strategy to eradicate microorganisms is photodynamic inactivation (PDI). In view of this, the present work aimed to synthesize and characterize chitosan nanoparticles (NPs) carrying curcumin (Cur NPs) to evaluate their effectiveness in the photodynamic inactivation (PDI) process. Cur NPs were characterized by atomic force microscopy, dynamic light scattering and Zeta potential. Furthermore, Cur-NPs achieved an encapsulation efficiency of 96%, and a slower release compared to free curcumin. To carry out the biological tests, the strains Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) were used. They were divided into two groups: (I) irradiated group and (II) non-irradiated group. The NPs internalization time was 1 hour. The irradiated groups of both strains were subjected to 1 hour of illumination with LED light, at a wavelength of 450 nm (blue) and an energy dose of 28.84 mW.cm-2. The concentrations of NPs Cur, 0.02 mg.mL-1; 0.04 mg.mL-1; 0.08 mg.mL-1; Polaxamer 407 at 25 mg.mL-1 and Chitosan + TPP at 1 mg.mL-1 for IFD were tested. To perform morphological analyzes of the strains after IFD, scanning electron microscopy (SEM) and Transmission Electron Cryomicroscopy (Cryo-ME) images were obtained. To evaluate the production of reactive oxygen species (ROS), the fluorescent marker dihydroethidium (DHE) was used and the production of ROS was monitored over time. Cur NPs showed photodynamic effects for the two strains tested, as well as inducing bactericidal effects at the highest concentrations tested. The SEM and Cryo-ME results showed that IFD promoted morphological changes in the structures of bacterial cell walls, lysing the cells. The results obtained indicate that Cur NPs have the potential for photoinactivation of bacteria and that these results are directly related to the production of ROS. Key words: nanoparticles, chitosan, curcumin, photodynamic inactivation, bacteria.
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spelling 2024-07-07T22:15:25Z2024-07-07T22:15:25Z2024https://repositorio.ufms.br/handle/123456789/8965For a long time, humanity has faced several diseases caused by bacterial infections. The discovery of antibiotics completely changed modern medicine and extended human life expectancy. However, abuse and misuse of antibiotics over the years is one of the factors that leads to bacterial resistance reaching alarming levels. The development of a new generation of anti-infective medicines using natural antimicrobial products is very promising. Another promising strategy to eradicate microorganisms is photodynamic inactivation (PDI). In view of this, the present work aimed to synthesize and characterize chitosan nanoparticles (NPs) carrying curcumin (Cur NPs) to evaluate their effectiveness in the photodynamic inactivation (PDI) process. Cur NPs were characterized by atomic force microscopy, dynamic light scattering and Zeta potential. Furthermore, Cur-NPs achieved an encapsulation efficiency of 96%, and a slower release compared to free curcumin. To carry out the biological tests, the strains Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) were used. They were divided into two groups: (I) irradiated group and (II) non-irradiated group. The NPs internalization time was 1 hour. The irradiated groups of both strains were subjected to 1 hour of illumination with LED light, at a wavelength of 450 nm (blue) and an energy dose of 28.84 mW.cm-2. The concentrations of NPs Cur, 0.02 mg.mL-1; 0.04 mg.mL-1; 0.08 mg.mL-1; Polaxamer 407 at 25 mg.mL-1 and Chitosan + TPP at 1 mg.mL-1 for IFD were tested. To perform morphological analyzes of the strains after IFD, scanning electron microscopy (SEM) and Transmission Electron Cryomicroscopy (Cryo-ME) images were obtained. To evaluate the production of reactive oxygen species (ROS), the fluorescent marker dihydroethidium (DHE) was used and the production of ROS was monitored over time. Cur NPs showed photodynamic effects for the two strains tested, as well as inducing bactericidal effects at the highest concentrations tested. The SEM and Cryo-ME results showed that IFD promoted morphological changes in the structures of bacterial cell walls, lysing the cells. The results obtained indicate that Cur NPs have the potential for photoinactivation of bacteria and that these results are directly related to the production of ROS. Key words: nanoparticles, chitosan, curcumin, photodynamic inactivation, bacteria.Há muito tempo a humanidade enfrenta diversas doenças ocasionas por infecções bacterianas. A descoberta dos antibióticos mudou completamente a medicina moderna e prolongaram a expectativa de vida humana. Contudo, abusos e utilização indevida de antibióticos ao longo dos anos é um dos fatores que leva a atingir a resistência bacteriana a níveis alarmantes. O desenvolvimento de uma nova geração de medicamentos anti-infecciosos utilizando produtos naturais antimicrobianos é bastante promissor, outra estratégia promissora para erradicar os microrganismos é a inativação fotodinâmica (IFD). Em vista disso, o presente trabalho teve como objetivo sintetizar e caracterizar nanopartículas (NPs) de quitosana carreando curcumina (NPs Cur) para avaliar sua eficácia no processo de inativação fotodinâmica (IFD). As NPs Cur foram caraterizadas por microscopia de força atômica, espalhamento dinâmico de luz e potencial Zeta. Além disso as NPs-Cur obtiveram uma eficiência de encapsulação de 96%, e uma liberação mais lenta em comparação a curcumina livre. Para a realização dos ensaios biológicos foram utilizadas as cepas Staphylococcus aureus (ATCC 25923) e Escherichia coli (ATCC 25922). Elas foram divididas em dois grupos: (I) grupo irradiado e (II) grupo não-irradiado. O tempo de internalização das NPs foi de 1 hora. Os grupos irradiados de ambas as cepas foram submetidos a 1 hora de iluminação com luz LED, no comprimento de onda de 450 nm (azul) e dose energética de 28,84 mW.cm-2. As concentrações de NPs Cur, 0,02 mg.mL-1; 0,04 mg.mL-1; 0,08 mg.mL-1; Polaxamer 407 a 25 mg.mL-1 e Quitosana + TPP a 1 mg.mL-1, para IFD foram testadas. Para realizar as análises morfológicas das cepas após a IFD, foram obtidas imagens de microscopia eletrônica de varredura (MEV) e Criomicroscopia Eletrônica de Transmissão (Crio-ME). Para avaliar a produção de espécies reativas de oxigênio (EROs) utilizou-se o marcador fluorescente dihydroethidium (DHE) e monitorou a produção de EROs ao longo do tempo. As NPs Cur apresentaram efeitos fotodinâmicos para as duas cepas testadas, bem como induziram efeitos bactericidas nas concentrações mais altas testadas. Os resultados de MEV e Crio-ME mostraram que a IFD promoveu mudanças morfológicas nas estruturas das paredes celulares das bacterianas, lisando as células. Os resultados obtidos indicam que as NPs Cur possuem potencial para fotoinativação de bactérias e que esses resultados estão diretamente relacionados a produção de EROs. Palavras chaves: nanopartículas, quitosana, curcumina, inativação fotodinâmica, bactérias.Fundação Universidade Federal de Mato Grosso do SulUFMSBrasilnanoparticulasquitosana inativação fotodinâmicaSíntese e caracterização de nanopartículas de quitosana carreando curcumina para avaliação de inativação fotodinâmicainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisAnderson Rodrigues Lima CairesLaís Fernandes Aguilerainfo:eu-repo/semantics/openAccessporreponame:Repositório Institucional da UFMSinstname:Universidade Federal de Mato Grosso do Sul (UFMS)instacron:UFMSORIGINALPGCM dissertação - Laís Fernandes Aguilera - versão final.pdfPGCM dissertação - Laís Fernandes Aguilera - versão final.pdfapplication/pdf4296795https://repositorio.ufms.br/bitstream/123456789/8965/-1/PGCM%20disserta%c3%a7%c3%a3o%20%20-%20La%c3%ads%20Fernandes%20Aguilera%20-%20vers%c3%a3o%20final.pdf38d001121921848df56b509662185e75MD5-1123456789/89652024-07-07 18:15:26.937oai:repositorio.ufms.br:123456789/8965Repositório InstitucionalPUBhttps://repositorio.ufms.br/oai/requestri.prograd@ufms.bropendoar:21242024-07-07T22:15:26Repositório Institucional da UFMS - Universidade Federal de Mato Grosso do Sul (UFMS)false
dc.title.pt_BR.fl_str_mv Síntese e caracterização de nanopartículas de quitosana carreando curcumina para avaliação de inativação fotodinâmica
title Síntese e caracterização de nanopartículas de quitosana carreando curcumina para avaliação de inativação fotodinâmica
spellingShingle Síntese e caracterização de nanopartículas de quitosana carreando curcumina para avaliação de inativação fotodinâmica
Laís Fernandes Aguilera
nanoparticulas
quitosana inativação fotodinâmica
title_short Síntese e caracterização de nanopartículas de quitosana carreando curcumina para avaliação de inativação fotodinâmica
title_full Síntese e caracterização de nanopartículas de quitosana carreando curcumina para avaliação de inativação fotodinâmica
title_fullStr Síntese e caracterização de nanopartículas de quitosana carreando curcumina para avaliação de inativação fotodinâmica
title_full_unstemmed Síntese e caracterização de nanopartículas de quitosana carreando curcumina para avaliação de inativação fotodinâmica
title_sort Síntese e caracterização de nanopartículas de quitosana carreando curcumina para avaliação de inativação fotodinâmica
author Laís Fernandes Aguilera
author_facet Laís Fernandes Aguilera
author_role author
dc.contributor.advisor1.fl_str_mv Anderson Rodrigues Lima Caires
dc.contributor.author.fl_str_mv Laís Fernandes Aguilera
contributor_str_mv Anderson Rodrigues Lima Caires
dc.subject.por.fl_str_mv nanoparticulas
quitosana inativação fotodinâmica
topic nanoparticulas
quitosana inativação fotodinâmica
description For a long time, humanity has faced several diseases caused by bacterial infections. The discovery of antibiotics completely changed modern medicine and extended human life expectancy. However, abuse and misuse of antibiotics over the years is one of the factors that leads to bacterial resistance reaching alarming levels. The development of a new generation of anti-infective medicines using natural antimicrobial products is very promising. Another promising strategy to eradicate microorganisms is photodynamic inactivation (PDI). In view of this, the present work aimed to synthesize and characterize chitosan nanoparticles (NPs) carrying curcumin (Cur NPs) to evaluate their effectiveness in the photodynamic inactivation (PDI) process. Cur NPs were characterized by atomic force microscopy, dynamic light scattering and Zeta potential. Furthermore, Cur-NPs achieved an encapsulation efficiency of 96%, and a slower release compared to free curcumin. To carry out the biological tests, the strains Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) were used. They were divided into two groups: (I) irradiated group and (II) non-irradiated group. The NPs internalization time was 1 hour. The irradiated groups of both strains were subjected to 1 hour of illumination with LED light, at a wavelength of 450 nm (blue) and an energy dose of 28.84 mW.cm-2. The concentrations of NPs Cur, 0.02 mg.mL-1; 0.04 mg.mL-1; 0.08 mg.mL-1; Polaxamer 407 at 25 mg.mL-1 and Chitosan + TPP at 1 mg.mL-1 for IFD were tested. To perform morphological analyzes of the strains after IFD, scanning electron microscopy (SEM) and Transmission Electron Cryomicroscopy (Cryo-ME) images were obtained. To evaluate the production of reactive oxygen species (ROS), the fluorescent marker dihydroethidium (DHE) was used and the production of ROS was monitored over time. Cur NPs showed photodynamic effects for the two strains tested, as well as inducing bactericidal effects at the highest concentrations tested. The SEM and Cryo-ME results showed that IFD promoted morphological changes in the structures of bacterial cell walls, lysing the cells. The results obtained indicate that Cur NPs have the potential for photoinactivation of bacteria and that these results are directly related to the production of ROS. Key words: nanoparticles, chitosan, curcumin, photodynamic inactivation, bacteria.
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publisher.none.fl_str_mv Fundação Universidade Federal de Mato Grosso do Sul
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