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Combined endo-periodontal lesions: microbiological and endotoxic evaluation

Grant number: 14/27365-1
Support type:Scholarships in Brazil - Master
Effective date (Start): July 01, 2016
Effective date (End): February 28, 2017
Field of knowledge:Health Sciences - Dentistry - Endodontics
Principal researcher:Brenda Paula Figueiredo de Almeida Gomes
Grantee:Rafaela Casadei Chapola
Home Institution: Faculdade de Odontologia de Piracicaba (FOP). Universidade Estadual de Campinas (UNICAMP). Piracicaba , SP, Brazil


The dental pulp and periodontal tissues are two anatomically distinct structures, but functionally interrelated, since both structures have embryological origin and concomitant formation. Although there is extensive literature regarding the microbiota associated with periodontal lesions, and endodontic infections separately, few studies have been dedicated to the investigation of microorganisms and their virulence factors, such as endotoxins (LPS) in combined endo-periodontal lesions. The objectives of this study are: a) to investigate the presence of microorganisms (Treponema denticola, Treponema socranskii, Gemella morbillorum, Tannerella forsythia, Aggregatibacter actinomycetemcomitans, Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Prevotella tannerae, Prevotella nigrescens, Fusobacterium nucleatum, Filifactor alocis, Parvimonas micra) in root canals and periodontal pockets of teeth with necrotic pulp and periodontal pockets greater than or equal to 6 mm, before and after chemical-mechanical preparation, and after the use of a temporary dressing, by Nested PCR method and by checkerboard, b ) to check the susceptibility of these microorganisms to the chemo-mechanical preparation and to the use of temporary dressing for 30 days, by counting of the colony forming units (CFU); c) To analyze the effect of chemo-mechanical preparation and the use of temporary dressing on the microorganisms of the associated periodontal pocket; d) To analyze the effect of the chemical-mechanical preparation and intracanal medication in the reduction of endotoxin levels of the root canal and associated periodontal pocket; e) To investigate possible associations between bacterial species detected and endotoxin with clinical signs and symptoms. Therefore, microbiological and endotoxin samples will be collected from root canals and periodontal pockets before and after chemical-mechanical preparation and after the use of temporary dressing; Means of transportation are used, specific culture and incubating to provide strict anaerobic growth, thus allowing the count of CFUs. The bacterial DNA from these samples will be extracted and then analyzed microbiologically by nested PCR and checkerboard. The endotoxins will be quantified by the turbidimetric LAL assay (Limulus amebocyte lysate). The data will be tabulated and statistically analyzed. (AU)

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