Systematic Approach to Identify Novel Antimicrobial and Antibiofilm Molecules from Plants' Extracts and Fractions to Prevent Dental Caries

Author(s)

Ribeiro, Sabrina M. ; Fratucelli, Erick D. O. ; Fernandes, Julia M. ; Bueno, Paula C. P. ; Cavalheiro, Alberto Jose ; Klein, Marlise, I

• Document type: Journal article
• Source: JOVE-JOURNAL OF VISUALIZED EXPERIMENTS; v. N/A, n. 169, p. 23-pg., 2021-03-01.

Abstract

Natural products provide structurally different substances, with a myriad of biological activities. However, the identification and isolation of active compounds from plants are challenging because of the complex plant matrix and time-consuming isolation and identification procedures. Therefore, a stepwise approach for screening natural compounds from plants, including the isolation and identification of potentially active molecules, is presented. It includes the collection of the plant material; preparation and fractionation of crude extracts; chromatography and spectrometry (UHPLC-DAD-HRMS and NMR) approaches for analysis and compounds identification; bioassays (antimicrobial and antibiofilm activities; bacterial "adhesion strength" to the salivary pellicle and initial glucan matrix treated with selected treatments); and data analysis. The model is simple, reproducible, and allows high-throughput screening of multiple compounds, concentrations, and treatment steps can be consistently controlled. The data obtained provide the foundation for future studies, including formulations with the most active extracts and/or fractions, isolation of molecules, modeling molecules to specific targets in microbial cells and biofilms. For example, one target to control cariogenic biofilm is to inhibit the activity of Streptococcus mutans glucosyltransferases that synthesize the extracellular matrix' glucans. The inhibition of those enzymes prevents the biofilm build-up, decreasing its virulence. (AU)

• FAPESP's process: 19/23175-7 - Formulations based on extracts and fractions from Casearia sylvestris for controlling cariogenic biofilm
• Grantee: Sabrina Marcela Ribeiro
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 13/07600-3 - CIBFar - Center for Innovation in Biodiversity and Drug Discovery
• Grantee: Glaucius Oliva
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 17/07408-6 - Evaluation of antimicrobial and antibiofilm activities of extracts and fractions from plants against oral pathogenic microorganisms
• Grantee: Sabrina Marcela Ribeiro
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 11/21440-3 - Infra-specific chemical variability and dynamics of phenolic and diterpenic secondary metabolites in Casearia sylvestris Sw (Salicaceae) from different Brazilian ecosystems: metabolomics and proteomics correlation
• Grantee: Paula Carolina Pires Bueno
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 12/21921-4 - Proteomics of two varieties of Casearia sylvestris Sw. (Salicaceae)
• Grantee: Paula Carolina Pires Bueno
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• FAPESP's process: 14/50926-0 - INCT 2014: biodiversity and natural products
• Grantee: Vanderlan da Silva Bolzani
• Support Opportunities: BIOTA-FAPESP Program - Thematic Grants