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Microorganisms and their virulence factors detection in saliva, pulp chamber and root canal of teeth associated with endodontic failure

Grant number: 15/19215-2
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): March 01, 2016
Effective date (End): February 28, 2017
Field of knowledge:Health Sciences - Dentistry - Endodontics
Principal Investigator:Brenda Paula Figueiredo de Almeida Gomes
Grantee:Priscila Amanda Francisco
Host Institution: Faculdade de Odontologia de Piracicaba (FOP). Universidade Estadual de Campinas (UNICAMP). Piracicaba , SP, Brazil
Associated scholarship(s):16/19743-1 - Metaproteomics analyses of root canal content of teeth with endodontic failure, BE.EP.MS


In the microbiota of secondary/ persistent infections predominate microorganisms resistant to endodontic procedures and able to survive in an environment with lack of space and nutrients, particularly Enterococcus faecalis. Coronal microleakage can be one of the causative agents of the failure of endodontic treatment, providing a constant source of saliva contaminants to the root canals. The aims of this study are: a) to study the saliva, pulp chamber and root canal microbiota composition of teeth with endodontic failure by checkerboard technique in order to investigate the similarity between microbiomes present in these 3 sites; b) to quantify the total bacterial load, Streptococcus spp and Enterococcus faecalis in saliva, pulp chamber and root canal, by Real Time - PCR; c) to quantify the levels of lipopolysaccharides (LPS) and lipoteichoic acid (LTA) present in the root canals; and to correlate them with the patients clinical and radiographic aspects. Twenty patients with the presence of apical periodontitis and the need for endodontic retreatment will be selected. Samples from the endotoxic contents from saliva, pulp chamber and root canal will be collected. The extracted DNA will be submitted to the Real Time polymerase chain reaction for quantification of the bacterial load, and of the levels of Streptococcus spp. and Enterococcus faecalis. The involved microbial species will be identified by the checkerboard using probes for 40 different selected endodontic pathogens. LPS and LTA will be quantified using the Limulus Amoebocyte Lysate (LAL) and Enzyme-Linked Immunosorbent Assay (ELISA) methods, respectively. Data will be tabulated and statistically analyzed by SPSS for Windows.

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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)
FRANCISCO, P. A.; DELBONI, M. G.; LIMA, A. R.; XIAO, Y.; SIQUEIRA, W. L.; GOMES, B. P. F. A.. Proteomic profile of root canal contents in teeth with post-treatment endodontic disease. International Endodontic Journal, v. 52, n. 4, p. 451-460, . (17/16516-7, 15/23479-5, 16/19743-1, 15/19215-2)
GOMES, BRENDA P. F. A.; FRANCISCO, PRISCILA AMANDA; GODOI JR, EDERALDO PIETRAFESA; ENDO, MARCOS SERGIO; BARBOSA-RIBEIRO, MARLOS; DELBONI, MARAISA GREGGIO; PECORARI, VANESSA G. A.. Identification of Culturable and Nonculturable Microorganisms, Lipopolysaccharides, and Lipoteichoic Acids From Root Canals of Teeth With Endodontic Failure. JOURNAL OF ENDODONTICS, v. 47, n. 7, p. 1075-1086, . (15/19215-2)
DELBONI, MARAISA G.; GOMES, BRENDA P. F. A.; FRANCISCO, PRISCILA A.; TEIXEIRA, FABRICIO B.; DRAKE, DAVID. Diversity of Enterococcus faecalis Genotypes from Multiple Oral Sites Associated with Endodontic Failure Using Repetitive Sequence-based Polymerase Chain Reaction and Arbitrarily Primed Polymerase Chain Reactionc. JOURNAL OF ENDODONTICS, v. 43, n. 3, p. 377-382, . (15/19215-2)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
FRANCISCO, Priscila Amanda. Microorganisms and their virulence factors detection in saliva, pulp chamber and root canal of teeth associated with endodontic failure. 2017. Master's Dissertation - Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba Piracicaba, SP.

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