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Microbiological analysis and quantification of endotoxins and proinflammatory cytokines from primarily infected root canals with apical periodontitis

Grant number: 10/13498-9
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): October 01, 2010
Effective date (End): February 28, 2014
Field of knowledge:Health Sciences - Dentistry - Endodontics
Principal Investigator:Brenda Paula Figueiredo de Almeida Gomes
Grantee:Daniel Rodrigo Herrera Morante
Host Institution: Faculdade de Odontologia de Piracicaba (FOP). Universidade Estadual de Campinas (UNICAMP). Piracicaba , SP, Brazil


The main virulence factor of gram-negative bacteria is represented by the release of endotoxin (LPS) presented in the outer layers of their cell walls. The accumulation of these bacterial components in the root canal and periradicular tissues should stimulate macrophages present in the area to produce cytokines such as IL-1 and TNF-± which are potent stimulators of bone resorption. The aim of this study is to isolate and identify gram-negative anaerobic species through biochemical methods and PCR from root canals of teeth with necrotic pulp and apical periodontitis, evaluating the effect of chemomechanical preparation (CMP) with 2% chlorhexidine gel and 17% EDTA, using or not ultrasound. The reduction of microorganisms and LPS, correlating the microbiota and endotoxin content with the expression of pro-inflammatory cytokines IL-1a, IL-1b, TNF± and PGE-2. Twenty-four root canals were selected with pulp necrosis and presence of periapical lesion. Endotoxins and microbiological samples will collect Samples were collected before and after CMP, before and after using 17% EDTA, with or without application of ultrasound, and finally after seven days of the first session. Samples for cytokines quantification will also collect by passing a paper point 1 mm beyond the apex, after the PQM and after 7 days, The microbiological samples will be diluted, plated and incubated for further isolation and identification. PCR will be performed using specific primers. A limulus amebocyte lysate (LAL) assay will be used to quantify endotoxins Cytokines collected from apex will be quantified using specific kits. The values obtained will be tabulated and statistically analyzed.

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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)
MARINHO, A. C. S.; MARTINHO, F. C.; GONCALVES, L. M.; RABANG, H. R. C.; GOMES, B. P. F. A.. Does the Reciproc file remove root canal bacteria and endotoxins as effectively as multifile rotary systems?. International Endodontic Journal, v. 48, n. 6, p. 542-548, . (13/02402-9, 10/13498-9, 10/17877-4, 13/17928-6, 10/19136-1, 11/09047-4)
HERRERA, D. R.; SILVA, E. J. N. L.; SANTOS, C. C.; ZAIA, A. A.; FERRAZ, C. C. R.; ALMEIDA, J. F. A.; GOMES, B. P. F. A.. Root canal content from primary endodontic infection and upregulation of gelatinases in fibroblast cells. International Endodontic Journal, v. 48, n. 12, p. 1168-1174, . (10/19136-1, 10/17877-4, 10/13498-9)
HERRERA, DANIEL R.; SANTOS, ZARINA T.; TAY, LIDIA Y.; SILVA, EMMANUEL J.; LOGUERCIO, ALESSANDRO D.; GOMES, BRENDA P. F. A.. Efficacy of different final irrigant activation protocols on smear layer removal by EDTA and citric acid. MICROSCOPY RESEARCH AND TECHNIQUE, v. 76, n. 4, p. 364-369, . (10/13498-9)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
MORANTE, Daniel Rodrigo Herrera. Análise microbiológica, de endotoxinas, de citocinas proinflamatórias e de metaloproteinases em infecções endodônticas primárias com lesões periapicais. 2014. Doctoral Thesis - Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba Piracicaba, SP.

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