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Effects of prenatal and perinatal probiotic supplementation on susceptibility to the development of experimental periodontitis through microbial challenge in the mice offspring

Abstract

The objective of this research will be to analyze how prenatal and perinatal probiotic (PROB) supplementation affects the development of experimental periodontal disease (PD) in the offspring of mice. 140 pregnant mice will be used, 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 offspring conceived by each of these groups will be divided into 2 subgroups: P1 (without induction of DP; n = 24) and P2 (with induction of DP; n = 24). After mating and confirmation of pregnancy, animals in the DPM and DPMP groups will receive gavages with Porphyromonas gingivalis w83 (5x109 colony forming units (CFU) / mL) for 37 days. In offspring animals (P2 subgroups), PD will be induced following the same protocol adopted for the DPM and DPMP groups, with gavages performed for 21 days. Animals in 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 before the animals were mated. Animals in the CM, CMP, DPM and DPMP groups will be euthanized on the 63rd. day of the experiment. Animals from subgroups P1 and P2 will be euthanized 63 days after birth. The following will be evaluated: 1) presence of B. lactis HN019 in the placenta of pregnant mice by means of real-time polymerase chain reaction (qPCR); 2) immunoinflammatory profile (IL-6, TNF-a, INF-y, IL-10, IL-1b, IL-8) present in placental tissues, periodontal tissues and amniotic fluid through immunoenzymatic assays (Luminex); 3) intestinal and oral microbial composition using qPCR; 4) immunohistochemical expression of Toll-like receptors (TLR-2 and TLR-4) in intestinal and placental tissues; 5) severity of inflammation present in the 2nd furcation region. upper molar by means of histomorphometric analyzes; 6) expression of E-cadherin proteins, junctional adhesion molecule (JAM), DNA methyltransferase 3 (DNMT3), acetylated histone H3 (AcH3) and acetylated histone H4 (AcH4) in periodontal and intestinal tissues by immunohistochemical analysis; 7) alveolar bone loss in the 2nd region. upper 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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