Experimental toothpaste containing glucanohydrolytic enzymes combined with Baccharis dracunculifolia extract: impact on the extracellular matrix of the carygenic biofilm

• Grant number: 22/15545-1
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: April 01, 2023
• Status: Discontinued
• Field of knowledge: Interdisciplinary Subjects
• Principal Investigator: Carolina Patrícia Aires Garbellini
• Grantee: Jéssica Silva Peixoto Bem
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated scholarship(s): 23/17451-7 - Immobilized enzymes in inorganic nanoparticles for cariogenic biofilm eradication, BE.EP.DR

Abstract

Dental caries is a biofilm-sugar dependent disease that destroys mineralized dental tissues. Its control should focus on a balanced diet and disorganization of the biofilm by daily brushing with dentifrices, which can be vehicles for active substances such as antimicrobials, aiming at controlling the cariogenic biofilm. However, the effect of these actives on the innermost portions of the biofilm is limited, due to the presence of the exopolysaccharide matrix. There are antibiofilm agents focused on the degradation of extracellular matrix polysaccharides, the main virulence factor of cariogenic biofilms, such as hydrolytic enzymes and Baccharis dracunculifolia extract. Nevertheless, the combined effect of these compounds has yet to be tested. Thus, the objective is to develop and evaluate the efficacy in biofilms of an experimental dentifrice containing these agents. In the first phase of the study, experimental dentifrice containing dextranase and mutanase enzymes encapsulated in a calcium-alginate matrix and Baccharis dracunculifolia extract will be developed. In the second phase, multispecies biofilms of Streptococcus mutans, Actinomyces naeslundii, and Streptococcus gordonii will be formed for 4 days on bovine enamel plates. From the 2nd day, the biofilms will be exposed to the treatments (n=9): a) NaCl 0.9%, negative control (NaCl); b) 0.12% chlorhexidine digluconate solution, positive control (CHX); c) Base dentifrice suspension containing 1,100 ppm of NaF (NaF); d) Toothpaste suspension containing mutanase, dextranase and B. dracunculifolia extract (MDBd); e) Dentifrice suspension containing mutanase and dextranase (MD); f) Dentifrice suspension containing B. dracunculifolia extract (Bd). The acidogenicity of biofilms, bacterial viability, the number of extracellular polysaccharides, and the loss of surface hardness of the specimens will be determined. In addition, confocal microscopy images will be performed to evaluate the ratio between the biovolume of bacteria and extracellular matrix and the three-dimensional structural organization of biofilms. The study will imply the development of new oral hygiene products from an ecological perspective of biofilms.