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Effect of mesenchymal stem cells from periodontal ligament with high and low osteogenic potential on bone repair

Grant number: 19/10076-0
Support type:Scholarships in Brazil - Master
Effective date (Start): August 01, 2019
Effective date (End): July 31, 2021
Field of knowledge:Health Sciences - Dentistry - Oral and Maxillofacial Surgery
Principal Investigator:Márcio Mateus Beloti
Grantee:Letícia Faustino Adolpho
Home 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

Abstract

Stem cell therapy is one of the strategies used in regenerative medicine and can be applied to regenerate bone tissue. Mesenchymal stem cells (MSCs) can be isolated from several body tissues, such as bone marrow, adipose tissue, umbilical cord, muscle and periodontal ligament. Our research group has evaluated strategies to promote bone regeneration in a rat calvaria defect model and has shown that cell locally injected was more effective than the use of biomaterials alone or in combination with cell therapy. Despite the efficacy of bone marrow MSCs, the procedure to obtain them is invasive, and alternative sources of MSCs are of interest to this research field. Considering that the periodontal ligament is easily harvested from extracted teeth, this tissue may be a source of MSCs as it is connective tissue rich in cells, including MSCs. In vitro and in vivo studies have demonstrated that these cells have the ability to differentiate into osteoblasts and to produce mineralized matrix. In addition, it was observed that MSCs from periodontal ligament may present distinct osteogenic potentials and those with high potential exhibit an early expression of Runx2. In this context, the aim of this study is to evaluate the capacity of MSCs from periodontal ligament with high and low osteogenic potential, and a mixture of both, to repair bone tissue. The cells will be obtained from the periodontal ligament of extracted third molars, in vitro characterized in terms of capacity for proliferation and osteoblastic differentiation and classified as MSCs with high or low osteogenic potential. Furthermore, bone defects with 5 mm in diameter will be created in calvaria of Wistar rats and after 2 weeks, cells with high and lo osteogenic potential, and a mixture of both, will be directly injected into bone defects (5x106 cells/defect). Four weeks post-injection, it will be evaluated: the bone formation, by histological analysis and microtomography, and the gene expression of Runx2, alkaline phosphatase, bone sialoprotein, osteocalcin, osteoprotegerin, Rankl, and Rank, by real-time PCR. The quantitative data will be submitted to adherence to normal curve test to determine the appropriate statistical test and the level of significance will be set at 5%. (AU)