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3D scaffolds functionalized with anti-fibronectin Aptamers on coagulum features. Ex vivo and In vivo studies.

Grant number: 18/12036-3
Support type:Regular Research Grants
Duration: February 01, 2019 - January 31, 2022
Field of knowledge:Health Sciences - Dentistry - Periodontology
Principal researcher:Daniela Bazan Palioto Bulle
Grantee:Daniela Bazan Palioto Bulle
Home Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Assoc. researchers:Paulo Tambasco de Oliveira

Abstract

In bone regeneration, blood is the first component to come into contact with the biomaterials used in periodontology. In this research we aimed to see the additional effects of anti-fibronectin (FN) Aptamers (short oligonucleotides capable of selectively binding to target protein) in blood clot formation and, additionally, bone formation. MTT will test in vitro cell viability of pre-osteoblastic cells (OSB) derived from rats (UMR-106 linage) on SCA with or without anti-FN Aptamers. The physiological clot (PhC) will be developed in an animal model (rat) on SCA with or without Aptamers, characterizing the ex vivo study. Also, PhC phenotyping will be characterized by flow cytometry and scanning electron microscopy. In another step, OSB will be cultured on the substrate (SCA, Aptamers and PhC), aiming to observe the effect of the substrate on the formation and accumulation of calcium and the 3D morphology of OSB and PhC by mineralization assay and confocal microscopy, respectively. The genetic interactions triggered by SCA, Aptamers and PhC association on OSB will be investigated by transcriptomic analysis using RNA-seq method. In in vivo study, we will verify this association in bone formation in rats. The experimental groups will be 1) negative control (without biomaterial in bone defect - C), 2) Control (SCA) and 3) SCA functionalized with Aptamers (SCA+APTantiFN). All will be investigated after 5 and 15 days to observe osteoblastic differentiation and bone neoformation. Two critical defects will be performed in the calvaria: one will be directed to bone density and volume formed analysis by Micro-CT, and other to transcriptomic analyze and histomorphometric processing. The genetic profile and the expression of Runx2 and alkaline phosphatase will be investigated using the RNA-seq and immunohistochemistry, respectively. These findings will be promising to observe the autocrine and paracrine signaling of blood clot formed on this biomaterial enriched with anti-FN Aptamer, being fundamental to discovery, in both the in vitro and in vivo studies, the effects of this association on osteogenesis. (AU)

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