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Susceptibility to the development of experimental periodontal disease in the offspring of mice with a history of prenatal and perinatal probiotic supplementation

Grant number: 19/12269-0
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): September 01, 2019
Status:Discontinued
Field of knowledge:Health Sciences - Dentistry - Periodontology
Principal researcher:Michel Reis Messora
Grantee:Pedro Henrique Felix Silva
Home Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated scholarship(s):20/14618-0 - Effects of prenatal and perinatal probiotic supplementation on the development of experimental periodontitis in pregnant mice and their offspring: microbiome analysis, BE.EP.DR

Abstract

The objective of this research will be to analyze how prenatal and perinatal probiotic supplementation (PROB) affects the development of experimental periodontal (PD) disease in the offspring of mice. 140 pregnant mice divided into 4 groups (n=35): CM (animals without PD and not treated with probiotics); DPM (animals with PD and not treated with probiotics); CMP (animals without PD and treated with probiotics) and DPMP (animals with PD and treated with probiotics). The progeny conceived by each of these groups will be divided into 2 subgroups: P1 (no PD induction; n=24) and P2 (with PD induction; n=24). After mating and confirmation of pregnancy, the animals of the DPM and DPMP groups will receive a silk bandage around the 2nd. upper molars and gavages with Porphyromonas gingivalis w83 5x109 colony forming units (CFU)/ml for 37 days. In the offspring of the offspring (subgroups P2), PD will be induced following the same protocol adopted for the DPM and DPMP groups, and the gavages were performed for 14 days. Animals of the CMP and DPMP groups will receive 1 x 109 CFU/mL of Bifidobacterium animalis subsp. lactis HN019 administered in water for 56 days starting 14 days prior to mating of the animals. Animals of the CM, CMP, DPM and DPMP groups will undergo euthanasia at the 56th. day of the experiment. Animals of subgroups P1 and P2 will undergo euthanasia 56 days after birth. The following will be evaluated: 1) presence of B. lactis HN019 in the placenta of pregnant mice by real-time polymerase chain reaction (qPCR); 2) immunoinflammatory profile (IL-6, TNF-±, INF-³, IL-10, IL-1², IL-8) present in placental tissues, periodontal tissues and amniotic fluid by immunoenzymatic assays (Luminex); 3) intestinal and oral microbial composition by of qPCR; 4) immunohistochemical expression of Toll-like receptors (TLR-2 and TLR-4) in the intestinal and placental tissues; 5) severity of the inflammation ratio present in the furcation region of the 2nd molar by histomorphometric analysis; 6) expression of E-cadherin proteins, Junctional Adhesion Molecule (JAM) and DNMT3 in periodontal and intestinal tissues by means of immunohistochemical assay; 7) alveolar bone loss ratio in the 2nd molar by means of x-ray computed microtomography (Micro-CT). The data obtained will be submitted to statistical analysis (p <0.05). (AU)

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