Effect of BMP-9 overexpression by CRISPR/Cas9 on mesenchymal stem cells on bone regeneration

• Grant number: 19/18550-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: November 01, 2019
• End date: October 31, 2021
• Field of knowledge: Health Sciences - Dentistry - Oral and Maxillofacial Surgery
• Principal Investigator: Gileade Pereira Freitas
• Grantee: Isabella Cristine dos Santos
• Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Bone is a tissue with high regeneration capacity, but in some situations, the extent of injury prevents tissue repair. In this scenario, cell therapy using bone marrow-derived mesenchymal stem cells (MSCs) has attracted the attention of several research groups because it is a promising alternative to existing treatments. Bone morphogenetic proteins (BMPs) are cytokines belonging to the transforming growth factor beta (TGF-b) family and are involved in various biological processes, including osteoblastic differentiation and bone formation. Among the several BMPs identified, bone morphogenetic protein 9 (BMP-9) is considered one of the most osteogenic and still little explored. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR-Cas9), becomes a viable alternative to be investigated in the repair of bone defects. Therefore, the present project aims to evaluate the effect of local injections of MSCs-BMP9 on bone tissue regeneration. For this, MSCs-BMP9 will be grown in growth medium for 10 days. For bone tissue regeneration, the MSCs-BMP9 will be injected directly into bone defects (5 mm in diameter) created in rat calvaria (5x106 cells/defect). At 4 weeks, the animals will be euthanized, and the samples will be evaluated by microtomographic analysis. Quantitative data will be subjected to the normal curve adherence test to determine the appropriate statistical test, and the significance level will be 5%. The results of this study may establish new parameters regarding cell therapy to regenerate bone tissue. (AU)