Effect of extracellular vesicles on osteoblast differentiation and bone regeneration induced by mesenchymal stem cells

• Grant number: 21/01499-5
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: June 01, 2021
• End date: January 12, 2023
• Field of knowledge: Health Sciences - Dentistry - Oral and Maxillofacial Surgery
• Principal Investigator: Adalberto Luiz Rosa
• Grantee: Gabriela Guaraldo Campos Tótoli
• Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 19/08568-2 - Investigation of the extracellular vesicles (VEs) role in the initiation, propagation, regeneration, and modeling of biological mineralization, AP.TEM

Abstract

Extracellular Vesicles (EVs) take part in sophisticated intercellular communication mechanisms relevant to the Regenerative Medicine field based on cellular therapy using Mesenchymal Stem Cells (MSCs) because these cells are able to generate and release EVs to the extracellular milieu. This study will test the hypothesis that EVs combined with MSCs induce the bone regeneration of rat calvaria defects by favoring the osteoblastic differentiation of MSCs and osteoblasts (OBs), the cells in charge of bone regeneration, through the BMPs, Wnt and ITGs signaling pathways. MSCs derived EVs will be harvested by ultracentrifugation and characterized by size, morphology and expression of EVs-marker proteins. The osteoblastic differentiation of MSCs and OBs will be evaluated by assaying the internalization of EVs into MSCs and OBs, the gene expression of bone markers and alkaline phosphatase activity. The participation of BMPs, Wnt and ITGs pathways will be evaluated by gene expression of a panel of genes followed by silencing of those higher expressed. The efficacy of the therapy with local injection of MSCs combined with Evs on bone formation in rat calvaria defects will be determined by microtomography and histological analyses. (AU)