Participation of agrin on osteoblast differentiation and bone tissue regeneration induced by mesenchymal stem cells in defects created in mouse calvaria

• Grant number: 21/03204-2
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: July 01, 2021
• End date: June 30, 2027
• Field of knowledge: Health Sciences - Dentistry - Oral and Maxillofacial Surgery
• Principal Investigator: Márcio Mateus Beloti
• Grantee: Maria Paula Oliveira Gomes
• Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 17/12622-7 - Cell therapy: potential of mesenchymal stem cells, VEGF-A and BMP-9 to regenerate bone tissue, AP.TEM
• Associated scholarship(s): 22/02461-4 - Role of Agrin and Wnt signaling inhibitor Sfrp4 on osteogenesis and odontogenesis, BE.EP.DD

Abstract

Cell therapy is one of the strategies of the regenerative medicine with potential for the treatment of large bone defects mainly using Mesenchymal Stem Cells (MSCs) and the study of molecules that act on these cells to favor bone regeneration is essential in this context. Among these molecules, the extracellular matrix protein agrin, which is involved in several biological processes, was detected in chondrocytes and it has been demonstrated its participation in skeletal system development. However, up to now, there is few information in the literature about the effects of agrin on osteoblast differentiation of MSCs and on bone formation induced by them, as well as the signaling pathways involved in these effects. Therefore, the aims of this study are to investigate: (1) in vitro the effect of overexpression and silencing of agrin by CRISPR/Cas9 on osteoblast differentiation of MSCs; (2) in vitro the participation of BMP, Wnt, Notch and Hippo-Yap/Taz signaling pathways on the effect of agrin on osteoblast differentiation of MSCs and (3) in vivo the effect of MSCs with overexpression and silencing of agrin by CRISPR/Cas9 on bone regeneration in defects created in mouse calvaria. The results of this study may generate important data about the function of agrin on bone tissue and, possibly, for the development of new therapeutic strategies in clinical situations involving bone regeneration. (AU)