Gymnema sylvestre: antiinflammatory and antimicrobial action on anaerobic bacteria of dental interest

• Grant number: 16/09959-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: December 01, 2016
• End date: November 30, 2017
• Field of knowledge: Health Sciences - Dentistry - Dental Clinics
• Principal Investigator: Luciane Dias de Oliveira
• Grantee: Manuela Maria Viana Miguel
• Host Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil

Abstract

The use of plant extracts have grown in several areas of health, however, in dentistry its use is still very limited , requiring scientific studies proving its anti- inflammatory and antimicrobial action, for their addition in toothpastes, mouthwashes, irrigation and intracanal medications and other drugs used to treat mouth infections. The objectives of this study are: a) to analyze the in vitro antimicrobial action of Gymnema sylvestre glycolic extract on planktonic culture and biofilms (monotypic) of Porphyromonas gingivalis and Prevotella intermedia; b) to analyze in vitro anti-inflammatory action of G. sylvestre extract in fibroblast (FMN-1) stimulated by lipopolysaccharide (LPS). For the planktonic form, the microdilution broth method is used, according to the Clinical and Laboratory Standards Institute (CLSI) to determine the minimum inhibitory concentrations (MIC) and minimal microbicidal (CMM). For biofilms standardized suspensions (107 cells/mL) are added to microplate wells and after 48 h at 37°C (anaerobic chamber) under agitation will be treated with the extract for 5 min and 24 hs. Will include a control group (physiological solution, NaCl). Then, biofilms must be broken with ultrasonic homogenizer, the suspensions are diluted and plated on Reinforced Clostridial Medium (RCM). After 48 or 72 h, the colony forming units are counted per milliliter (CFU/mL) and the values will be converted to log10. To evaluate the anti-inflammatory action of the extract, fibroblasts (FMN-1) are cultivated in microplates for 24 h, after that, are added to different concentrations of the extract and lipopolysaccharide (LPS) from Escherichia coli (1 ug/ mL / well). After incubation for 24 h (37°C and 5% CO2), the supernatant will be collected to evaluate pro-inflammatory cytokines by ELISA assay. The results will be statistically analyzed by ANOVA and Tukey Test (5%). (AU)