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Previous issue date: 2012-02-28 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Genistein (GEN), isoflavone contained in soybeans, shows activity against a
large number of cancers, including skin cancer. However, to be used topically it is
essential the association of GEN in an appropriate formulation. The aim of this study
was the development and characterization of solid lipid nanoparticles (SLN) contends
genistein for topical application. A bioanalytical method was developed and validated
for GEN quantification in skin layers with High Performance Liquid Chromatographer
(HPLC) with UV detection. The SLN was obtained with glyceryl behenate,
polysorbate 80, sorbitan trioleate and different amounts of cetylpyridinium chloride
(CPC) (0.5 - 0.05%). The characteristics in terms of particle size, size distribution,
zeta potential, entrapment efficiency and drug recovery were evaluated. In vitro
release, passive and iontophoretic, skin permeation studies were performed.
Cytotoxicity studies were carried out in melanoma cells (B16F10) with free drug and
SLN with and without GEN. The analytical method was linear in the concentration
range of 0.1 to 60 mg/mL. Limit of quantification was 100 ng/mL for both skin
layers. Recovery of the drug ranged from 95.57 to 97.57%. The method was able to
analyze the GEN without suffering interference from endogenous skin
components. SLN loaded with GEN showed positive surface (+ 23 mV) and average
size of 343 nm. Particles were obtained with high entrapment efficiency (93%) and
drug load was 5.45%. In vitro release studies demonstrated that the release of GEN
from SLN occurs in two stages with a large amount of drug released within the first
six hours, followed by a slow release of the remaining drug in the lipid matrix of SLN
in the following hours. When administered in the skin, during in vitro passive
permeation studies, the SLN increased retention of GEN in stratum corneum and
remaining skin compared with free GEN. After iontophoresis application in
formulation of SLN containing GEN thirteen times more GEN was retained on stratum
corneum and three times more drug was retained on remaining skin. In cellular
cytotoxicity studies SLN favored the increase of interaction of drug with the cells and
the cytotoxicity was concentration-dependent. Thus, GEN loaded SLN increased drug
skin permeation and retention and shows to be a potential formulation for topical
application. / A genisteína (GEN), isoflavona contida nos grãos da soja, apresenta atividade
contra um grande número de tipos de câncer, incluindo o câncer de pele. No
entanto, para ser usada topicamente é fundamental que a GEN esteja associada a
uma formulação adequada. O objetivo deste trabalho foi desenvolver e caracterizar
nanopartículas lipídicas sólidas (NLS) contendo genisteína para aplicação tópica.
Para a quantificação da genisteína nas diferentes camadas da pele foi desenvolvida
e validada uma metodologia bioanalítica em cromatografia líquida de alta eficiência
(CLAE) com detecção no UV. As NLS foram obtidas com behenato de glicerila,
polissorbato 80, trioleato de sorbitano e tensoativo catiônico cloreto de cetilpiridínio
(CPC) em diferentes quantidades (0,5 – 0,05%). As NLS foram caracterizadas
quanto ao tamanho, PdI, potencial zeta, eficiência de encapsulação e recuperação.
Foram realizados estudos in vitro de liberação e permeação cutânea passiva e
iontoforética. Estudos de citotoxicidade foram realizados em linhagem de melanoma
(B16F10) com fármaco livre e NLS com e sem GEN. O método de quantificação do
fármaco mostrou-se linear na faixa de concentração de 0,1 a 60 μg/mL. O limite de
quantificação foi de 100 ng/mL para ambas as camadas da pele. A recuperação do
fármaco variou entre 95,57 a 97,57%. Ainda, o método foi capaz de analisar a GEN
sem sofrer interferência dos componentes endógenos da pele. As NLS carregadas
com fármaco apresentaram carga superficial positiva (+ 23 mV) e tamanho médio de
343 nm. Também foram obtidas partículas com alta eficiência de encapsulação
(93%) e carga de fármaco de 5,45%. Os estudos de liberação in vitro demonstraram
que a liberação da GEN a partir das NLS ocorre em duas fases, com uma grande
quantidade de fármaco liberada nas seis primeiras horas, seguida por uma liberação
lenta do restante do fármaco da matriz lipídica das NLS nas horas seguintes.
Quando administrada na pele, nos estudos de permeação passiva in vitro, as NLS
aumentaram a retenção da GEN tanto no estrato córneo quanto na pele
remanescente em comparação à administração da GEN livre. Após a aplicação da
iontoforese na formulação de NLS contendo GEN, treze vezes mais GEN ficou retida
no EC e três vezes mais fármaco ficou retido na pele remanescente. Nos estudos de
citotoxicidade, as NLS favoreceram o aumento da interação do fármaco com as
células e a citotoxicidade foi concentração-dependente. Deste modo, a
encapsulação da GEN em NLS aumentou a permeação e retenção do fármaco na
pele, demonstrando assim, potencial para aplicação tópica.
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.bc.ufg.br:tde/3041 |
| Date | 28 February 2012 |
| Creators | Silva, Lorena Maione |
| Contributors | Lima, Eliana Martins, Taveira, Stephânia Fleury |
| Publisher | Universidade Federal de Goiás, Programa de Pós-graduação em Ciências Farmacêuticas (FF), UFG, Brasil, Faculdade Farmácia - FF (RG) |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | Portuguese |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFG, instname:Universidade Federal de Goiás, instacron:UFG |
| Rights | http://creativecommons.org/licenses/by-nc-nd/4.0/, info:eu-repo/semantics/openAccess |
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