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Evaluation of cell behavior and response of subcutaneous connective tissue of isogenic mice induced by endodontic sealers

Grant number: 13/21180-7
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): January 01, 2014
Effective date (End): December 31, 2014
Field of knowledge:Health Sciences - Dentistry
Principal researcher:Léa Assed Bezerra da Silva
Grantee:Leopoldo Ulian Azevedo
Home Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated scholarship(s):14/08671-4 - Evaluation of cell behavior induced by endodontic sealers, BE.EP.IC


The aim of this study is to evaluate cell behavior and response of subcutaneous connective tissue of isogenic mice induced by sealers Real Seal XT1 and Sealapex Xpress. The cell viability and proliferation will be assessed after exposure of cells to cement Sealapex Spress (group I) and Real Seal XT1 (group II). As a control, cells will be exposed to bacterial endotoxin (LPS) (Group III) and to the DMEM-c culture (Group IV). Cell viability of the suspension of RAW 264.7 macrophage lineage will be evaluated in different groups by MTT colorimetric assay (Sigma). Cell viability in groups I and II will be expressed in percentage and compared to groups III and IV (controls). Cell proliferation will be determined by immunostaining the Ki-67 nuclear antigen expressed only by cells during the cell cycle and not by cells in G0. The result will be expressed as the total number of cells per well. The proportion of cells that exhibit changes in the cytoplasmic membrane will be determined by flow cytometry from 1 to 10 days. The readings will be held in flow cytometry and apoptosis will be expressed in percentage compared to groups III and IV. In order to analyze the response in the subcutaneous tissue of isogenic mice will be made of Teflon matrices to obtain specimens of the filling materials, which ones will be introduced in the lumbar region of each animal within the subcutaneous tissue. Will be used 70 isogenic strain BALB/c male, aged 6 to 8 weeks old, weighing 15 to 20 grams, which will be divided into 8 groups (experimental and control). At the end of each of the experimental periods (7, 21 and 63 days), a portion of the subcutaneous connective tissue will be set aside and submitted to histotechnical processing routine. In peripheral tissues to the test material, the density will be observed number of inflammatory cells, the immunohistochemical test for detection of interleukin-6 will be observed too, as any possible phenomenon encountered. Numerical data will be analyzed using the appropriate test according to the nature of the data. The significance level will be 0,05.

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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)
SILVA, L. A. B.; AZEVEDO, L. U.; CONSOLARO, A.; BARNETT, F.; XU, Y.; BATTAGLINO, R. A.; CANADAS, P. S.; HOLANDA DE OLIVEIRA, KATHARINA MORANT; SILVA, R. A. B.. Novel endodontic sealers induce cell cytotoxicity and apoptosis in a dose-dependent behavior and favorable response in mice subcutaneous tissue. CLINICAL ORAL INVESTIGATIONS, v. 21, n. 9, p. 2851-2861, . (13/21180-7)

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