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Master's Dissertation
DOI
https://doi.org/10.11606/D.58.2020.tde-04102022-083622
Document
Author
Full name
Leonardo Lobo Ribeiro Cavalcante
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2019
Supervisor
Committee
Corona, Silmara Aparecida Milori (President)
Rastelli, Alessandra Nara de Souza
Silva, Cláudia Helena Lovato da
Watanabe, Evandro
Title in Portuguese
Conjugado de nanopartículas de quitosana com ftalocianina de cloro alumínio: síntese, caracterização e fotoinativação de biofilmes de S. mutans
Keywords in Portuguese
Biofilme
Ftalocianina de cloro alumínio
MEV
Nanocarreadores
Nanopartículas de quitosana
Terapia fotodinâmica antimicrobiana
Abstract in Portuguese
A terapia fotodinâmica antimicrobiana (aPDT) utilizando ftalocianina de cloro alumínio (ClAlPc) tem alto poder oxidativo, sendo alternativa para o controle de biofilmes cariogênicos, principalmente ao serem administradas em veículos de liberação adequados. Este estudo teve como objetivo caracterizar a ClAlPc encapsulada em nanopartículas de quitosana (NQ) e avaliar suas propriedades antimicrobianas contra biofilmes de S. mutans. As NQ foram preparadas por geleificação iônica e os estudos de caracterização incluíram o tamanho das partículas, índice de polidispersão (IPd), potencial zeta, estabilidade acelerada, espectro de absorção e quantificação de ClAlPc. Os biofilmes foram crescidos por 48 h em blocos de dentina bovina (37°C, sob microaerofilia). Para a aPDT a ClAlPc na concentração final de 8 µM foi combinada com o laser de diodo (InGaAlP) com 660 nm e 100 J/cm2. A toxicidade do aPDT foi verificada por fototoxicidade no escuro. A atividade antimicrobiana foi verificada por UFC. Os biofilmes foram analisados por microscopia eletrônica de varredura (MEV). O número de bactérias viáveis foi analisado pelos testes ANOVA e Tukey HSD (α = 0,05). A caracterização revelou que as nanopartículas de ClAlPc foram encontradas em escala nanométrica, com propriedades fotofísicas e fotoquímicas adequadas. O aPDT mediada pelo nanoconjugado ClAlPc + NQ mostrou redução significativa na viabilidade de S. mutans (<1log10 UFC / mL) em comparação ao controle negativo PBS (p <0,05). A aPDT mediada por ClAlPc foi semelhante ao PBS (p> 0,05). A MEV revelou que os biofilmes tratados com aPDT ClAlPc + NQ reduziram a quantidade de bactérias na dentina. As células encontravam-se isoladas ou dispostas em cadeias curtas e foram encontrados formatos irregulares de S. mutans. As nanopartículas de ClAlPc são consideradas estáveis e o aPDT mediada pelo nanoconjugado ClAlPc + NQ foi eficaz contra o biofilme de S. mutans.
Title in English
Conjugate of chitosan nanoparticles with chloroaluminium phthalocyanine: synthesis, characterization and photoinactivation of S. mutans biofilm
Keywords in English
Antimicrobial photodynamic therapy
Biofilm
Chitosan nanoparticles
Chloroaluminium phthalocyanine
Nanocarriers
SEM
Abstract in English
Antimicrobial photodynamic therapy (aPDT) using chloroaluminium phthalocyanine (ClAlPc) has high oxidative power, being an alternative for the control of biofilms, especially when this photosensitizer is administered in an appropiate release vehicle. This study aimed to characterize the ClAlPc encapsulated in chitosan nanoparticles (CSNPs), and evaluate its antimicrobial properties against S. mutans biofilms. CSNPs were prepared by ion gelation and characterization studies included particle size, polydispersion index (IPd), zeta potential, accelerated stability, absorption spectrum and ClAlPc quantification. Bovine dentin blocks were incubated with S. mutans. Biofilm was grown for 48h at 37°C under microaerophilic. 8 µM ClAlPc was combined with diode laser (InGaAlP) at 660 nm and 100 J/cm2. The aPDT toxicity was verified by dark phototoxicity. The antimicrobial activity was verified by CFU. Biofilms were analyzed by scanning electron microscopy (SEM). The number of viable bacteria was analyzed by ANOVA and Tukey HSD tests (α=0.05). The characterization revealed that the ClAlPc nanoparticles were found in nanometerscale with adequate photophysical and photochemical properties. The aPDT mediated by ClAlPc + CSNPs nanoconjugate showed a significant reduction in the viability of S. mutans (<1log10 CFU/mL) compared to the negative control (PBS, p<0.05). The aPDT mediated by ClAlPc was similar to PBS (p>0.05). SEM revealed that the biofilms treated with aPDT ClAlPc + CSNPs reduced the amount of bacteria on the dentin. Cells were arraged as single or in shorted chains. Irregular shapes of S. mutans were found. ClAlPc nanoparticles are considered stable and aPDT mediated by ClAlPc + CSNPs nanoconjugate was effective against S. mutans biofilm.
 
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Publishing Date
2022-10-05
 
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