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Antimicrobial and anti-inflammatory activity, cytotoxicity and genotoxicity of different natural extracts

Grant number: 15/08776-3
Support type: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
Home Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil
Assoc. researchers:Flaviana Bombarda de Andrade


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)

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
CAMARGO, SAMIRA ESTEVES AFONSO; DE OLIVEIRA, LUCIANE DIAS. Curcuma longaL. (turmeric),Rosmarinus officinalisL. (rosemary), andThymus vulgarisL. (thyme) extracts aid murine macrophages (RAW 264.7) to fightStreptococcus mutansduring in vitro infection. Archives of Microbiology, v. 202, n. 8 JUN 2020. Web of Science Citations: 0.
DE OLIVEIRA, JONATAS RAFAEL; VIEGAS, DAIANE DE JESUS; REQUIA MARTINS, ANA PAULA; TALGE CARVALHO, CLAUDIO ANTONIO; SOARES, CRISTINA PACHECO; AFONSO CAMARGO, SAMIRA ESTEVES; CARDOSO JORGE, ANTONIO OLAVO; DE OLIVEIRA, LUCIANE DIAS. Thymus vulgaris L. extract has antimicrobial and anti-inflammatory effects in the absence of cytotoxicity and genotoxicity. ARCHIVES OF ORAL BIOLOGY, v. 82, p. 271-279, OCT 2017. Web of Science Citations: 4.
DE OLIVEIRA, JONATAS RAFAEL; FIGUEIRA, LEANDRO WAGNER; SPER, FABIA LUGLI; MECCATTI, VANESSA MARQUES; AFONSO CAMARGO, SAMIRA ESTEVES; DE OLIVEIRA, LUCIANE DIAS. Thymus vulgaris L. and thymol assist murine macrophages (RAW 264.7) in the control of in vitro infections by Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. IMMUNOLOGIC RESEARCH, v. 65, n. 4, p. 932-943, AUG 2017. Web of Science Citations: 2.

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