Desenvolvimento e caracterização de nanoestruturas poliméricas para encapsulação de um novo derivado de antraquinona de origem biotecnológica
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Data
2019-02-19
Autores
Amantino, Camila Fernanda
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Universidade Estadual Paulista (Unesp)
Resumo
Os materiais nanoestruturados têm potencial diferente em comparação com outros materiais em maior escala, resultando em diferentes características e propriedades físico-químicas. Inicialmente foram preparadas nanoemulsões (NE) contendo corante vermelho (CV) um derivado de antraquinona, produzido pelo cultivo do fungo Talaromyces minioluteus em mesa incubadora rotativa, no entanto as NE com corante vermelho (CV/NE) não apresentaram características físico-químicas desejadas (tamanho de partícula ≤ 250 nm, índice de polidispersividade ≤0,3), partindo-se então para a utilização de nanocápsulas poliméricas (NC) como sistemas carreadores para este ativo. Foram preparadas amostras de NC vazias (Vazia/NC), com corante vermelho (CV/NC), com corante vermelho associado a ftalocianina de cloro alumínio (CV_ClAlPc/NC) e somente com ftalocianina de cloro alumínio (ClAlPc/NC). Estes nanomateriais foram caracterizados e apresentaram características físico- químicas adequadas, com tamanho de partícula < 250 nm, índice de polidispersividade ≤ 0,3 e potencial Zeta predominantemente negativo. As análises de estabilidade acelerada apresentaram um tempo vida de armazenamento > 3 meses para todas as NC. As análises de microscopia por força atômica (MFA) realizadas para CV/NC e CV_ClAlPc/NC mostram uma morfologia esférica para ambas as NC e tamanho de partícula de 104 nm para CV/NC e 211nm para CV_ClAlPc/NC, as análises por diferença de fase mostraram que a superfície da partícula é homogênea, ou seja, composta somente pelo poli-láctico-co-glicólico (PLGA) e também é possível afirmar que não há presença de ativo livre no meio das NC. Foram realizadas análises por espectroscópicas de absorção e de emissão de fluorescência UV-visível as quais não mostraram alterações nas propriedades fotofísicas no estado estacionário do CV livre e da ClAlPc livre em comparação com CV/NC , CV_ClAlPc/NC e ClAlPc/NC, foi possível estabelecer um método analítico para a quantificação do corante baseado na curva de correlação entre a absorção máxima em função da concentração de CV na faixa de 1,0 a 20 μg/ml onde foi obtido um coeficiente de correlação da ordem de 0,999. Foram quantificadas todas as NC as quais apresentaram uma taxa de incorporação entre 60 -70 %, e eficiência de encapsulação de 100%, resultado comprovado pela análise de diferença de fase por microscopia de força atômica (MFA). Foram realizadas também análises de tempo de vida de fluorescência, as quais apresentarem ajuste multiexponencial para CV/DMSO_ACN, CV/NC e CV_ClAlPc/NC, apresentando dois tempos de vidas quando excitados em 470 nm e ajuste monoexponencial para CV_ClAlPc/NC e ClAlPc/NC, apresentanto apenas um tempo de vida quando excitados em 640 nm. Os ensaios de viabilidade celular mostram ausência de citotoxidade para CV livre e para as NC, com uma viabilidade maior que 93%. Os resultados obtidos neste trabalho indicam que as nanocápsulas poliméricas apresentaram grande potencial como nanoestrutura para encapsulação do CV e quando associado a ClAlPc. As NC apresentaram estabilidade, características físico-químicas desejadas, e alta eficiência de encapsulação dos ativos, além de não apresentarem citotoxidade para células saudáveis, dando assim suporte para estudos futuros para diversas aplicações biológica e/ou biomédica.
Nanostructured materials have different potential compared to other materials on a larger scale, resulting in different physico-chemical characteristics and properties. Initially NE were prepared containing red dye (RD) an anthraquinone derivative produced by cultivating the Talaromyces minioluteus fungus on rotary incubator table. However, the RD/NE have not shown the desired physical-chemical characteristics, starting with the use of nanocapsules as carrier systems for this active. Empty NC samples (blank / NC), with red dye (RD / NC), with red dye associated with phthalocyanine chlorine aluminum (RD_ClAlPc / NC) and with phthalocyanine chlorine aluminum (ClAlPc / NC) were prepared. These nanomaterials were characterized and presented adequate physico-chemical characteristics, with particle size <250 nm, polydispersity index ≤ 0.3 and Zeta potential predominantly negative. The accelerated stability analyzes showed a shelf life> 3 months for all NCs. Atomic force microscopy (MFA) analyzes performed for RD / NC and RD_ClAlPc / NC show a spherical morphology for both NC and particle size of 104 nm for RD / NC and 211nm for RD_ClAlPc / NC. The difference analysis of phase showed that the particle surface is homogeneous, that is, composed only of polylactic-co-glycolic acid (PLGA) and it is possible to affirm that there is no free active in the NC medium. UV-visible spectroscopic absorption and emission spectroscopic analyzes were performed which did not show changes in the free-stationary photophysical properties of free RD and free ClAlPc compared to RD / NC, RD_ClAlPc / NC and ClAlPc / NC. It was possible to establish an analytical method for the quantification of the dye based on the correlation curve between the maximum absorption as a function of the concentration of RD in the range of 1.0 to 20 μg / mL where a correlation coefficient of the order of 0,999 . It was quantified all the NCs which had an incorporation rate between 60-70%, and encapsulation efficiency of 100%, a result evidenced by phase difference analysis by atomic force microscopy (MFA). Fluorescence life-time analyzes were performed, which present a multiexponential adjustment for RD / DMSO_ACN, RD / NC and RD_ClAlPc / NC, with two lifetimes when excited at 470 nm and monoexponential adjustment for RD_ClAlPc / NC and ClAlPc / NC , presenting only a lifetime when excited at 640 nm. Cell viability assays show absence of cytotoxicity for free RD and for NC, with viability greater than 93%. The results obtained in this work indicate that the polymeric nanocapsules presented great potential as nanostructure for RD encapsulation and when associated with ClAlPc. NC showed stability, desired physical-chemical characteristics, and high efficiency of encapsulation of the active, besides not presenting cytotoxicity to healthy cells, thus supporting future studies for several biological and / or biomedical applications.
Nanostructured materials have different potential compared to other materials on a larger scale, resulting in different physico-chemical characteristics and properties. Initially NE were prepared containing red dye (RD) an anthraquinone derivative produced by cultivating the Talaromyces minioluteus fungus on rotary incubator table. However, the RD/NE have not shown the desired physical-chemical characteristics, starting with the use of nanocapsules as carrier systems for this active. Empty NC samples (blank / NC), with red dye (RD / NC), with red dye associated with phthalocyanine chlorine aluminum (RD_ClAlPc / NC) and with phthalocyanine chlorine aluminum (ClAlPc / NC) were prepared. These nanomaterials were characterized and presented adequate physico-chemical characteristics, with particle size <250 nm, polydispersity index ≤ 0.3 and Zeta potential predominantly negative. The accelerated stability analyzes showed a shelf life> 3 months for all NCs. Atomic force microscopy (MFA) analyzes performed for RD / NC and RD_ClAlPc / NC show a spherical morphology for both NC and particle size of 104 nm for RD / NC and 211nm for RD_ClAlPc / NC. The difference analysis of phase showed that the particle surface is homogeneous, that is, composed only of polylactic-co-glycolic acid (PLGA) and it is possible to affirm that there is no free active in the NC medium. UV-visible spectroscopic absorption and emission spectroscopic analyzes were performed which did not show changes in the free-stationary photophysical properties of free RD and free ClAlPc compared to RD / NC, RD_ClAlPc / NC and ClAlPc / NC. It was possible to establish an analytical method for the quantification of the dye based on the correlation curve between the maximum absorption as a function of the concentration of RD in the range of 1.0 to 20 μg / mL where a correlation coefficient of the order of 0,999 . It was quantified all the NCs which had an incorporation rate between 60-70%, and encapsulation efficiency of 100%, a result evidenced by phase difference analysis by atomic force microscopy (MFA). Fluorescence life-time analyzes were performed, which present a multiexponential adjustment for RD / DMSO_ACN, RD / NC and RD_ClAlPc / NC, with two lifetimes when excited at 470 nm and monoexponential adjustment for RD_ClAlPc / NC and ClAlPc / NC , presenting only a lifetime when excited at 640 nm. Cell viability assays show absence of cytotoxicity for free RD and for NC, with viability greater than 93%. The results obtained in this work indicate that the polymeric nanocapsules presented great potential as nanostructure for RD encapsulation and when associated with ClAlPc. NC showed stability, desired physical-chemical characteristics, and high efficiency of encapsulation of the active, besides not presenting cytotoxicity to healthy cells, thus supporting future studies for several biological and / or biomedical applications.
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Corantes biotecnológicos, Nanocápsulas, Fotoquímica, Fotofísica, Espectroscopia., biotechnological dyes, Nanocapsules, Photochemistry, Photophysic, Spectroscopy