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Evaluation of the effect of endodontic and periodontal therapy on infectious and inflammatory contents in teeth with combined endo-periodontal lesions

Grant number: 17/14912-2
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): October 01, 2017
Effective date (End): May 31, 2021
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
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

Abstract

The dental pulp and the periodontium are two anatomically distinct but functionally interrelated structures, since both structures have an embryological origin and concomitant formation. Although there is an extensive literature on the microbiota associated with periodontal lesions and endodontic lesions separately, few studies have investigated infectious and inflammatory contents in teeth with combined Endo-Periodontal Lesions (EPL). LPS, or endotoxins, is the main virulence factor of Gram-negative bacteria present on the outer membrane of the bacterial cell envelope. The accumulation of this bacterial component in the root canal and its exit to the periodontal tissues stimulates an antigen-antibody reaction by the cells of the host's immune system, generating an inflammatory response at the periapical level. This reaction is characterized by the expression of chemical mediators and enzymes, such as proinflammatory cytokines and metalloproteinases (MMPs), in addition to the production of an important neuropeptide, involved in inflammatory phenomena, substance P. The objectives of the present study will be: to study the composition of the microbiota present in root canals and periodontal pockets of teeth with combined endo-periodontal lesions using the DNA-DNA Hybridization checkerboard technique and Nested-PCR; to verify the susceptibility of these microorganisms to endodontic and periodontal procedures and to the use of intracanal dressing for 30 days, through counting Colony Forming Units (CFU´s); to analyze the effect of the chemical-mechanical preparation and the use of intracanal dressing on the reduction of endotoxin levels, on the production of interleukin 1 alpha (IL-1±) 1 beta (IL-1²), Tumor Necrosis Factor alpha (TNF-±), on the levels of MMP-2, MMP-3, MMP-8, MMP-9 and MMP-13 and on the reduction of substance P concentration in the root canal and the associated periodontal pocket; correlate these findings with clinical signs and symptoms. Therefore, samples will be collected from the periodontal pockets and root canals for the quantification of endotoxins, proinflammatory cytokines, metalloproteinases and substance P and for microbiological analysis. . It will be used appropriated transport medium and specific culture medium and incubation to provide the growth of the strict anaerobic microorganisms, allowing the counting of the Colony Forming Units (CFU). The bacterial DNA from these samples will be extracted and then analyzed microbiologically through Checkerboard DNA-DNA Hybridization and Nested-PCR. The quantification of the endotoxins will be done by turbidimetric test, using the reagent LAL (Limulus Amebocytes Lysate). For the quantification of cytokines, MMP's and substance P the ELISA test (Enzyme-Linked Immunosorbent) will be performed. The data obtained will be tabulated and statistically analyzed. (AU)

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Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
CHAPOLA, Rafaela Casadei. Evaluation of the effect of endodontic and periodontal therapy against infectious content in teeth with necrotic pulps compromised by chronic periodontal disease. 2021. Doctoral Thesis - Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba Piracicaba, SP.

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