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Master's Dissertation
DOI
https://doi.org/10.11606/D.23.2023.tde-19062023-124817
Document
Author
Full name
Larissa Tavares Sampaio Silva
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2023
Supervisor
Committee
Roscoe, Marina Guimarães (President)
Boaro, Letícia Cristina Cidreira
Braga, Roberto Ruggiero
Rodrigues, Marcela Charantola
Title in Portuguese
Desenvolvimento de partículas de fosfato dicálcico dihidratado carregadas com clorexidina
Keywords in Portuguese
Agentes antimicrobianos
Clorexidina
Fosfato de Cálcio
Materiais dentários
Remineralização
Abstract in Portuguese
Os objetivos do presente estudo foram sintetizar, caracterizar e avaliar a atividade antibacteriana contra Streptococcus mutans de partículas de fosfato dicálcico dihidratado (DCPD) carregadas com diacetato de clorexidina (CHX). Para isso, foram sintetizados três grupos de partículas: DCPD: Partículas de DCPD sem a adição de CHX, DCPD/CHX-1: Partículas de DCPD carregadas com 1% de CHX, e DCPD/CHX-5: Partículas de DCPD carregadas com 5% de CHX. Na primeira etapa, as partículas sintetizadas foram caracterizadas para confirmar a composição da fase de fosfato de cálcio formada, o teor de clorexidina incorporado à partícula, bem como a morfologia e as dimensões. Para isso, foram realizadas análises por espectrofotometria UV/Visível (UV/Vis), espectroscopia no infravermelho por transformada de Fourier (FTIR), análise elementar, espalhamento de luz laser, e microscopia eletrônica de varredura (MEV). Na segunda etapa, as partículas foram avaliadas quanto à atividade antibacteriana, por meio da determinação da Concentração Inibitória Mínima (CIM). Os dados foram submetidos à análise de variância (ANOVA), complementada pelo teste de Dunnett, com nível de significância de 5%. A presença da CHX nas partículas DCPD/CHX foi confirmada quimicamente pelo método de FTIR e pela análise elementar e quantificada por espectrofotometria UV/Vis. A análise elementar demonstrou presença de maior teor de carbono (5,48 %) para o DCPD/CHX-1, seguido pelo grupo DCPD/CHX-5 (3,50%). O DCPD sem CHX não apresentou porcentagem significativa de carbono. Com a espectrofotometria UV/Vis, foi possível quantificar apenas o grupo DCPD/CHX-1, o qual apresenta 0,12 g de CHX a cada 1 g de DCPD. Através do espalhamento de luz laser (D50) foram detectadas partículas com 11 m de diâmetro em DCPD, 14 m em DCPD/CHX-1 e 12 m em DCPD/CHX-5. Nesta análise também foram mensuradas as áreas superficiais: 0,73 , 0,57 e 0,63 m2/g, respectivamente. A análise em MEV revelou a presença de partículas menores com formato arredondado (clorexidina) adsorvidas sobre a superfície da placa (DCPD) em DCPD/CHX-1. Apenas as partículas DCPD/CHX-1 apresentaram efetiva atividade antimicrobiana contra Streptococcus mutans, mesmo em baixas concentrações (CIM: 0,0038 mg/mL). Com base no êxito obtido na síntese de partículas DPCD/CHX, foi possível concluir que apenas as partículas de DCPD carregadas com 1% de CHX exibiram efetiva atividade antibacteriana contra Streptococcus mutans e, portanto, apresentam potencial para serem incorporadas à materiais odontológicos e fornecer concomitante atividade antibacteriana e propriedade remineralizante.
Title in English
Development of chlorhexidine-loaded calcium phosphate dihydrate particles
Keywords in English
Antimicrobial agents
Calcium phosphate
Chlorhexidine
Dental materials
Remineralization
Abstract in English
This study aimed to synthesize, characterize, and evaluate the antibacterial activity against Streptococcus mutans of calcium phosphate dihydrate particles (DCPD) loaded with chlorhexidine diacetate (CHX). Three groups of particles were synthesized: DCPD: DCPD particles without the addition of CHX, DCPD/CHX-1: DCPD particles loaded with 1% CHX, and DCPD/CHX-5: DCPD particles loaded with 5% CHX. The synthesized particles were characterized to confirm the calcium phosphate phase, the chlorhexidine content incorporated, as well as the morphology and dimensions. Therefore, analyzes were carried out by UV/Visible spectrophotometry (UV/Vis), Fourier transform infrared spectroscopy (FTIR), elemental analysis, laser light scattering, and scanning electron microscopy (SEM). After characterization, the antibacterial activity was evaluated, by determining the Minimum Inhibitory Concentration (MIC). The data were analyzed using analysis of variance (ANOVA), followed by Dunnett's test, with a significance level of 5%. The presence of CHX in the DCPD/CHX particles was chemically confirmed by the FTIR method and by elemental analysis and quantified by UV/Vis spectrophotometry. The elemental analysis showed the presence of a higher carbon content (5.48%) for the DCPD/CHX-1, followed by the DCPD/CHX-5 group (3.50%). The DCPD with no incorporation of CHX did not show a significant percentage of carbon. With UV/Vis spectrophotometry, it was possible to quantify only the DCPD/CHX-1 group, which presents 0.12 g of CHX for 1 g of DCPD. Through laser light scattering (D50), particle sizes were detected: 11 m diameter in DCPD, 14 m in DCPD/CHX-1, and 12 m in DCPD/CHX-5. In this analysis, the surface area was also measured: 0.73, 0.57, and 0.63 m2/g, respectively. The SEM analysis revealed the presence of smaller particles with spherical shape (chlorhexidine) adsorbed on the surface of the plates (DCPD) in DCPD/CHX-1. Only DCPD/CHX-1 particle showed effective antimicrobial activity against Streptococcus mutans, even at low concentrations (MIC: 0.0038 mg/mL). Based on these results, it was possible to conclude that the only DCPD particles loaded with 1% CHX exhibited effective antibacterial activity against Streptococcus mutans and, therefore, have the potential to be incorporated into dental materials and provide concomitant antibacterial activity and remineralizing properties.
 
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Publishing Date
2023-06-27
 
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