| Grant number: | 15/08776-3 |
| Support Opportunities: | Regular Research Grants |
| Duration: | October 01, 2015 - September 30, 2017 |
| Field of knowledge: | Biological Sciences - Microbiology - Applied Microbiology |
| Principal Investigator: | Luciane Dias de Oliveira |
| Grantee: | Luciane Dias de Oliveira |
| Host Institution: | Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil |
| Associated researchers: | Flaviana Bombarda de Andrade |
Abstract
The use of plant extracts in Dentistry is still very restricted and because of it, more scientific studies more specific are necessary to promote their inclusion in mouthwashes, toothpastes, irrigants and intracanal medications, ointments, etc., in order to direct the therapeutic indications. This study proposes to verify in vitro: a) antimicrobial activity of natural extracts of Cynara scolymus (artichoke), Cordia verbenacea (whaling herb), Gymnema sylvestre (gimena), Thymus vulgaris (thyme) and Juglans regia (walnut) on planktonic culture and biofilms of Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans, Pseudomonas aeruginosa, Porphyromonas gingivalis, Porphyromonas endodontalis, Parvimonas micra, Fusobacterium nucleatum and Prevotella intermedia; b) anti-inflammatory action in cultures of macrophages (RAW 264.7) and human gingival fibroblasts (FMN-1), after stimulation with Escherichia coli lipopolysaccharide (LPS), with analysis of the production of pro- and anti-inflammatory cytokines and nitric oxide; c) cytotoxic and genotoxic activities in macrophages (RAW 264.7) and gingival fibroblasts (FMN-1). For planktonic forms, broth microdilution method will be used, according to the Clinical and Laboratory Standards Institute (CLSI) to determine the minimum inhibitory concentrations (MIC) and minimum microbicidal concentration (MMC). For biofilms, standardized suspensions (107 cells/mL) will be added to microplate wells and after 48 h at 37°C, under agitation (75 rpm), they will be treated with different natural extracts (contact time: 5 min and 24 h). Control groups (saline solution, nystatin and chlorhexidine) will be included. Then, biofilms will be disaggregated and the suspensions will be seeded Reinforced Clostridial medium agar for anaerobic bacteria, Sabouraud-dextrose agar (yeast) or Brain Heart Infusion agar (other bacteria). After 48 hours at 37°C, the colony forming units per milliliter (CFU/mL) will be counted. The anti-inflammatory activity will be evaluated in cultures of macrophages and gingival fibroblasts stimulated with LPS and treated with the extracts. After 24 h, the supernatant will be removed and the quantification of nitric oxide, by Griess method, and proinflammatory cytokines (IL-1², TNF-±, IL-6, IL-12, IL-17) and anti-inflammatory (IL-10), by immunoenzymatic ELISA test, will be performed. Cytotoxicity will be analyzed by mitochondrial activity (MTT assay) and genotoxicity by micronucleus test. Statistical analysis will be performed by ANOVA and Tukey's test with level of significance of 5%. (AU)
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