Antimicrobial and anti-inflammatory activity of Hamamelis virginiana glycolic extract

• Grant number: 14/24713-9
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: February 01, 2015
• End date: January 31, 2016
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Luciane Dias de Oliveira
• Grantee: Nathália Bustamante Coura Ronconi Costa
• 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 in dentistry is still very restricted, so that more specific scientific studies are needed to promote their inclusion in mouthwashes, toothpastes, irrigation and intracanal medications, among others, in order to direct the therapeutic indications. The purpose of this study is to evaluate in vitro: a) antimicrobial activity of Hamamelis virginiana extract on planktonic culture and biofilms (monotypic) of Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans and Pseudomonas aeruginosa, b) anti- inflammatory action on macrophages stimulated by lipopolissacarídeo (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 under agitation (75 rpm) will be treated with the extract (contact time : 5 min). Will include a control group (saline). Then, biofilms must be broken with ultrasonic homogenizer, the suspensions are diluted and plated on Sabouraud's dextrose agar (yeast) or Brain Heart Infusion agar (bacteria). After 48 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, macrophages (RAW 264.7) 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 nitric oxide by the Griess method. The results will be statistically analyzed by ANOVA and Tukey Test (p d 0.05).