Expression of extracellular matrix production genes and antimicrobial resistance genes in situ cariogenic biofilms

• Grant number: 24/21446-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: March 01, 2025
• End date: December 31, 2025
• Field of knowledge: Health Sciences - Dentistry
• Principal Investigator: Marlise Inêz Klein Furlan
• Grantee: Pedro Canto Bueno
• Host Institution: Faculdade de Odontologia de Piracicaba (FOP). Universidade Estadual de Campinas (UNICAMP). Piracicaba , SP, Brazil
• Associated research grant: 21/06801-1 - Extracellular matrix: from biology to strategies for controlling cariogenic biofilms, AP.JP2

Abstract

Dental caries is a disease mediated by biofilm and modulated by dietary sugars (chiefly sucrose). It is still necessary to better understand this dysbiotic biofilm and whether approaches to reestablish eubiosis are efficacious. Thus, the current project is linked to the execution of an in situ experiment (PhD Fellowship FAPESP 2024/05965-9) that is part of an ongoing grant (FAPESP 2021/06801-1). The in situ study will be conducted with 16 participants (model with palatal apparatus containing dental blocks, crossover, and washout periods). Four solutions will be prepared and provided to the participants in each experimental group per phase, being: (1) sucrose and starch (cariogenic diet); (2) proline (potential prebiotic); (3) sodium nitrate (potential prebiotic) and (4) water (control). Several biofilm analyses will be performed to characterize the extracellular matrix, the microbiota, the metabolic pathways being expressed, the mineral content of the biofilm fluid, and its structure. The current scientific initiation proposal aims to evaluate the expression of extracellular matrix production genes and antimicrobial resistance genes in situ cariogenic biofilms. To achieve this aim, 64 in situ biofilm samples will be subjected to RNA isolation; the mRNA will be converted to cDNA. The cDNA samples will be evaluated via qPCR using specific primers for extracellular matrix synthesis and maintenance genes (e.g., gtfs, ftfs, gbps, dexA, fruA) and antimicrobial resistance genes (e.g., mecA, tetA, rpoB). The quantitative data will be analyzed using descriptive and inferential statistics (¿=0.05) and will be compared with the literature.