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Microbial and immunological characterization of endo-periodontal lesions

Grant number: 14/01655-3
Support Opportunities:Scholarships abroad - Research
Effective date (Start): September 15, 2014
Effective date (End): July 26, 2015
Field of knowledge:Health Sciences - Dentistry - Endodontics
Principal Investigator:Brenda Paula Figueiredo de Almeida Gomes
Grantee:Brenda Paula Figueiredo de Almeida Gomes
Host Investigator: Bruce J Paster
Host Institution: Faculdade de Odontologia de Piracicaba (FOP). Universidade Estadual de Campinas (UNICAMP). Piracicaba , SP, Brazil
Research place: Forsyth Institute, United States  

Abstract

Although there is an extensive literature on the microbiota associated with periodontal lesions and with endodontic lesions separately, few studies have been dedicated to the investigation of the combined lesions, such as the endo-periodontal lesions (EPL). The objectives of this study are: 1) To identify the presence, levels and proportions of reference bacterial species in root canals (RC) and periodontal pockets (PP) in teeth with EPL; 2) To analyze the microbiological profile of samples collected from RC and PP; 3) To investigate and quantify the presence of endotoxin (LPS) in RC and PP; 4) To quantify IL -1² and TNF-± proinflammatory cytokines in RC and PP; 5) To correlate specific microorganisms (MO), endotoxin levels and cytokines with clinical and radiographic findings of the involved teeth. Fifteen patients with teeth presenting necrotic pulp, periapical lesion and associated periodontal pockets, featuring endo-periodontal involvement, were selected, to assess the microbiological content, endotoxin levels and inflammatory potential. Samples were collected with sterile/ apyrogenic paper points from the RC and PP and frozen for further analysis. MO of these two sites will be identified by molecular techniques (Nested PCR and Checkerboard) and the microbial diversity and the relative abundance of MO found will be investigated by sequencing the samples using the Illumina MiSeq platform. The quantification of endotoxin in both sites will be accomplished through the LAL assay and the quantification of cytokines by ELISA. Before statistical analysis, the data obtained from sequencing will be assessed using the "pipeline" QIIME. All results, as well as the clinical and radiographic aspects of the involved teeth will be tabulated in Excel and subsequently analyzed with specific softwares. (AU)

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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)
GOMES, BRENDA P. F. A.; BERBER, VANESSA B.; KOKARAS, ALEXIS S.; CHEN, TSUTE; PASTER, BRUCE J.. Microbiomes of Endodontic-Periodontal Lesions before and after Chemomechanical Preparation. JOURNAL OF ENDODONTICS, v. 41, n. 12, p. 1975-1984, . (14/01655-3)
GOMES, BRENDA P. F. A.; BERBER, VANESSA B.; MARINHO, ARIANE C. S.; LOUZADA, LIDIANE M.; ARRUDA-VASCONCELOS, RODRIGO; PASSINI, MAICON R. Z.; LOPES, ERICA M.; PECORARI, VANESSA G. A.; CHEN, TSUTE; PASTER, BRUCE J.. Chemomechanical preparation influences the microbial community and the levels of LPS, LTA and cytokines in combined endodontic-periodontal lesions: A clinical study. JOURNAL OF PERIODONTAL RESEARCH, . (17/25242-8, 14/01655-3, 19/10755-5, 15/23479-5, 19/19300-0)

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