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Doctoral Thesis
DOI
https://doi.org/10.11606/T.9.2022.tde-25082022-164027
Document
Author
Full name
Magner Pessoa da Silva
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2022
Supervisor
Committee
Fabi, João Paulo (President)
Cuccovia, Iolanda Midea
Ong, Thomas Prates
Souza, Valdomiro Vagner de
Title in Portuguese
Preparação e caracterização de nanocarreadores de beta-lactose à base de pectina e lisozima
Keywords in Portuguese
Auto-organização
Biopolímero
Calorimetria de varredura diferencial
Lactose
Nanoencapsulação
Abstract in Portuguese
O objetivo deste trabalho foi preparar e caracterizar nanocarreadores via auto-organização a partir da pectina de citros e lisozima para o encapsulamento da β-lactose. Foram estudadas três condições de interação entre os biopolímeros variando a razão molar pectina/lisozima (3:1, 2:1, 1:1, 1:2 e 1:3), o pH e o tempo de aquecimento. A confirmação da interação foi determinada por espectroscopia no infravermelho por transformada de Fourier (FTIR) e por calorimetria de varredura diferencial (DSC). Os espectros de infravermelho evidenciaram que ligações de hidrogênio foram as principais forças envolvidas na formação dos nanocarreadores e sugeriram a ausência de β-lactose livre na superfície das nanopartículas. Os termogramas evidenciaram que as nanopartículas formadas na presença de β-lactose têm maior estabilidade térmica do que as nanopartículas sem β-lactose. Para ambas as formulações estudadas, na presença e na ausência de β-lactose, a formação das nanopartículas ocorreu entre os valores de pKa e ponto isoelétrico (pI) da pectina e lisozima, respectivamente, sendo a melhor razão de interação pectina/lisozima 1:2, em pH 10, a 80 ºC por 30 min. As nanopartículas foram formadas via auto-organização e todos as partículas apresentaram distribuição de tamanho homogênea, formato esférico, diâmetro inferior a 100 nm e carga superficial negativa. A morfologia e o tamanho das partículas pouco alteraram com a incorporação da -lactose. A eficiência de encapsulação (EE) da β-lactose foi superior a 96% para as concentrações estudadas. Ensaios preliminares in vitro, em células epiteliais de câncer de cólon (HCT-116), evidenciaram que as nanopartículas formadas são capazes de adentrar no meio intracelular, possivelmente, por via endocitose.
Title in English
Preparation and characterization of beta-lactose nanocarriers based on pectin and lysozyme
Keywords in English
Biopolymer
Differential scanning calorimetry
Lactose
Nanoencapsulation
Self-assembly
Abstract in English
This work aimed to prepare and characterize nanocarriers via self-assembly using citrus pectin and lysozyme for β-lactose encapsulation. Three interaction conditions between the biopolymers were studied, varying the pectin/lysozyme molar ratio (3:1, 2:1, 1:1, 1:2 and 1:3), pH and heating time. Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) determined the interaction's confirmation. The infrared spectra showed that hydrogen bonds were the main forces involved in the formation of nanocarriers and suggested the absence of free β-lactose on the surface of the nanoparticles. The thermograms showed that nanoparticles formed in the presence of β-lactose have greater thermal stability than nanoparticles without β-lactose. For both formulations studied, in the presence and absence of lactose, the formation of nanoparticles occurred between the pKa and isoelectric point (pI) values of pectin and lysozyme, respectively, with the best pectin/lysozyme interaction molar ratio 1:2, at pH 10, at 80 °C for 30 min. Nanoparticles were formed via self-assembly, and all particles presented homogeneous size distribution, spherical shape, diameter less than 100 nm, and negative surface charge. The morphology and size of the particles changed little with the incorporation of β-lactose. The encapsulation efficiency (EE) of β-lactose was higher than 96% for the concentrations studied. Preliminary in vitro assays in colon cancer epithelial cells (HCT-116) showed that the nanoparticles formed are capable of entering the intracellular medium, possibly via endocytosis.
 
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Publishing Date
2022-12-19
 
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