Síntese, caracterização e efeitos fotodinâmicos de curcumina encapsulada em nanoparticulas poliméricas
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Data
2016-06-24
Autores
Trigo Gutierrez, Jeffersson Krishan [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O objetivo deste estudo foi formular e avaliar os efeitos fotodinâmicos da curcumina (CUR) encapsulada em nanopartículas poliméricas (NP) em culturas plactônicas de Candida albicans, Streptococcus mutans e Staphylococcus aureus resistente à meticilina e em biofilme misto formado por essas três espécies. NP de CUR aniônicas e catiônicas foram sintetizadas utilizando-se polímero poli-ácido lático (PLA) e sulfato de dextran (DEX). Para caracterização dessa solução, foram determinadas as propriedades fisicoquímicas (tamanho, polidispersão e potencial zeta) e realizados testes de eficiência de encapsulamento, espectro de absorção, fotoestabilidade (fotodegradação da CUR) e atividade fotodinâmica (fotodegração de reagente e scavenger de oxigênio singleto). Em seguida, os efeitos da Terapia Fotodinâmica antimicrobiana (aPDT) mediada pela CUR-NP contra culturas planctônicas e biofilme misto cultivado in vitro por 48 horas, foram avaliados por meio de quantificação de colônias (UFC/mL). Cada avaliação foi realizada em triplicata em três ocasiões distintas e comparada com CUR livre. Os dados obtidos foram submetidos à análise estatística descritiva e inferencial (α=0,05). Após a síntese as CUR-NP apresentaram propriedades nanomêtricas farmacotécnicas adequadas , com valores médios de tamanho, polidispersão e potencial zeta 237,9; 0,08 e -35,2 mV; 239; 0,20 e +32 mV respectivamente para as CUR-NP aniônica e catiônica. Foi observada uma eficiência de encapsulamento de 67 e 64,6 % para as CUR-NP aniônicas e catiônicas. Os sinais de máxima absorção de luz para as CUR-NP e CUR foram pr[oximos da faixa UVvisível, também foi observada a degradação das formulações em diferentes períodos de iluminação. A liberação de CUR das NP aniônicas e catiônicas foi de 96% após 48 horas de estudo. Também foi observada maior atividade fotodinâmica após 5 minutos de irradiação tanto para as CUR-NP quanto para a CUR livre. aPDT mediada por CUR livre demostrou melhoor eficácia antimicrobiana contra os três micro-organismos tanto em culturas planctônicas quanto em biofilme misto de três espécies comparada com a CUR-NP aniônica mediadora de aPDT uma vez que a CUR livre erradicou os três micro-organismos em cultura planctônica(p< 0,001) e promoveu diminuição da viabilidade do biofilme misto, observou-se maior efetividade sobre C. albicans (p≤0,026). Enquanto a CUR-NP aniônica associada a luz azul promoveu a diminuição da viabilidade dos três micro-organismos em cultura planctônicas tendo maior efeito sobre S. mutans (p< 0,001), além disso, não foi observada diminuição da viabilidade do biofilme misto(p≥0,129). A CUR-NP catiônica apresentou efeito antimicrobiano mesmo na ausência de luz em culturas planctônicas dos três micro-organismos (p< 0,001) e biofilme misto, sendo a bactéria S. mutans o mais suscetível à formulação (p< 0,001). A formulação sintetizada foi solúvel em água e homogênea e a eficácia fotodinâmica antimicrobiana em culturas planctônicas foi inferior comparada com a CUR livre.
The aim of this study was to formulate and evaluate the photodynamic effect of curcumin (CUR) encapsulated in polymeric nanoparticles (NP) on planktonic cultures of Candida albicans, Streptococcus mutans and methicillin-resistant Staphylococcus aureus and on multispecies biofilm formed by these microorganisms. Anionic and cationic CUR-NP were synthesized using poly-lactic acid (PLA), and dextran sulfate (DEX). To characterize this solution, the physicochemical properties (size, polydispersity and zeta potential) were determined and tests of encapsulation efficiency, absorption spectrum, photostability (photodegradation of CUR) and photodynamic activity (photobleaching of singlet oxygen reagent and scavenger) were also performed. Next, antimicrobial effects of CUR-NP-mediated Photodynamic Therapy (PDT) against planktonic cultures and mixed biofilms grown in vitro for 48 hours were assessed by quantification of colonies (CFU/mL), Each evaluation were carried out in triplicate on three separate occasions and compared to free CUR. Data were submited to descritve and inferential statistical analize (α = 0.05). After synthesis CUR-NP had appropriate pharmacotechnical nanometric properties, mean values of size, polydispersity, and zeta potential were 237.9; 0.08 and -35.2 mV; 239; 0.20 and +32 mV, respectively for anionic and cationic CUR-NP. Encapsulation efficiency of 67 and 64.6% for anionic and cationic CURNP were observed. Peaks absorption for CUR-NP and CUR were close to UV-visible range, was also observed degradation of the formulations at different times of lighting. The release of CUR from anionic and cationic NP was 96% after 48 hours of study. It was also observed greater photodynamic activity after 5 minutes of irradiation for both CUR-NP and for the free CUR. Free CUR mediated aPDT demonstrated better antimicrobial effects against the three microorganisms both planktonic cultures as for mixed biofilms of three species compared with aPDT mediated by anionic CUR-NP, because aPDT mediated by free CUR erradicated the three microorganisms in the planktonic culture ( p <0.001) and promoted decrease in viability of the mixed biofilm, there was more effective on C. albicans (p≤0,026). While anionic CURNP associated with blue light promoted reduction of viability of the three planktonic microorganisms in culture with better effect on S. mutans (p <0.001) Moreover, there was no decrease in viability of mixed biofilm by anionic CUR-NP (p≥0,129). Cationic CUR-NP showed antimicrobial effect even in light absence of against planktonic cultures of the three micro-organisms (p <0.001), and mixed biofilm, bacteria S. mutans was the most susceptible to formulation (p <0.001). The synthesized formulation improved solubility in water and stability CUR-NP were homogeneous and showed antimicrobial photodynamic efficacy was superior in planktonic cultures was lower compared to the free CUR.
The aim of this study was to formulate and evaluate the photodynamic effect of curcumin (CUR) encapsulated in polymeric nanoparticles (NP) on planktonic cultures of Candida albicans, Streptococcus mutans and methicillin-resistant Staphylococcus aureus and on multispecies biofilm formed by these microorganisms. Anionic and cationic CUR-NP were synthesized using poly-lactic acid (PLA), and dextran sulfate (DEX). To characterize this solution, the physicochemical properties (size, polydispersity and zeta potential) were determined and tests of encapsulation efficiency, absorption spectrum, photostability (photodegradation of CUR) and photodynamic activity (photobleaching of singlet oxygen reagent and scavenger) were also performed. Next, antimicrobial effects of CUR-NP-mediated Photodynamic Therapy (PDT) against planktonic cultures and mixed biofilms grown in vitro for 48 hours were assessed by quantification of colonies (CFU/mL), Each evaluation were carried out in triplicate on three separate occasions and compared to free CUR. Data were submited to descritve and inferential statistical analize (α = 0.05). After synthesis CUR-NP had appropriate pharmacotechnical nanometric properties, mean values of size, polydispersity, and zeta potential were 237.9; 0.08 and -35.2 mV; 239; 0.20 and +32 mV, respectively for anionic and cationic CUR-NP. Encapsulation efficiency of 67 and 64.6% for anionic and cationic CURNP were observed. Peaks absorption for CUR-NP and CUR were close to UV-visible range, was also observed degradation of the formulations at different times of lighting. The release of CUR from anionic and cationic NP was 96% after 48 hours of study. It was also observed greater photodynamic activity after 5 minutes of irradiation for both CUR-NP and for the free CUR. Free CUR mediated aPDT demonstrated better antimicrobial effects against the three microorganisms both planktonic cultures as for mixed biofilms of three species compared with aPDT mediated by anionic CUR-NP, because aPDT mediated by free CUR erradicated the three microorganisms in the planktonic culture ( p <0.001) and promoted decrease in viability of the mixed biofilm, there was more effective on C. albicans (p≤0,026). While anionic CURNP associated with blue light promoted reduction of viability of the three planktonic microorganisms in culture with better effect on S. mutans (p <0.001) Moreover, there was no decrease in viability of mixed biofilm by anionic CUR-NP (p≥0,129). Cationic CUR-NP showed antimicrobial effect even in light absence of against planktonic cultures of the three micro-organisms (p <0.001), and mixed biofilm, bacteria S. mutans was the most susceptible to formulation (p <0.001). The synthesized formulation improved solubility in water and stability CUR-NP were homogeneous and showed antimicrobial photodynamic efficacy was superior in planktonic cultures was lower compared to the free CUR.
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Palavras-chave
Fotoquimioterapia, Fungo, Bactérias, Nanoparticulas, Photochemotherapy, Yeast, Bacteria, Nanoparticles