Expression of the extracellular matrix protein agrin during osteoblastic differentiation

• Grant number: 17/20349-9
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: January 01, 2018
• End date: December 31, 2018
• Field of knowledge: Health Sciences - Dentistry - Oral and Maxillofacial Surgery
• Principal Investigator: Adalberto Luiz Rosa
• Grantee: Denise Weffort
• Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Agrin is an extracellular matrix protein expressed in several tissues and involved in many biological processes. Among them, Agrin is expressed in cartilage and has a relevant role in the skeletal development. However, up to now, the expression of Agrin in osteoblasts has been underexplored. Therefore, the aims of this study are: (1) to evaluate the time-course gene and protein expression of Agrin during osteoblastic differentiation and (2) to investigate a possible correlation between Agrin expression and the stage of osteoblastic differentiation. We will use three mouse cell culture models: (1) mesenchymal stem cells from bone marrow, (2) calvarial cells and (3) immortalized osteoblastic cell line MC3T3-E1. These cells will be cultured under osteogenic conditions and the gene expression of Agrin will be evaluated at 3, 5, 7, 10 and 14 days by real-time PCR. The protein expression of Agrin will be evaluated at 5, 7 and 10 days by Western blot. Additionally, in order to investigate a possible correlation between Agrin expression and the level of osteoblastic differentiation, the gene expression of osteocalcin will be evaluated at the same periods used for Agrin and the in situ alkaline phosphatase activity will be assayed at 10 days. The numerical data will be submitted to the test of adherence to the normal distribution to determine the statistical analysis that should be used. The results of this study may generate relevant data to drive future investigations on the role of Agrin in bone tissue and, consequently, to the development of novel therapeutic strategies for bone regeneration. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

SOUZA, ALANN THAFFARELL PORTILHO; LOPES, HELENA BACHA; OLIVEIRA, FABIOLA SINGARETTI; WEFFORT, DENISE; FREITAS, GILEADE PEREIRA; ADOLPHO, LETICIA FAUSTINO; FERNANDES, ROGER RODRIGO; ROSA, ADALBERTO LUIZ; BELOTI, MARCIO MATEUS. The extracellular matrix protein Agrin is expressed by osteoblasts and contributes to their differentiation. Cell and Tissue Research, v. 386, n. 2, JUL 2021. (16/14171-0, 17/20349-9)

SOUZA, ALANN THAFFARELL PORTILHO; FREITAS, GILEADE PEREIRA; LOPES, HELENA BACHA; TOTOLI, GABRIELA GUARALDO CAMPOS; TARONE, ADRIANA GADIOLI; MAROSTICA-JUNIOR, MARIO ROBERTO; ROSA, ADALBERTO LUIZ; BELOTI, MARCIO MATEUS. Jabuticaba peel extract modulates adipocyte and osteoblast differentiation of MSCs from healthy and osteoporotic rats. JOURNAL OF BONE AND MINERAL METABOLISM, v. 39, n. 2, SEP 2020. (15/13320-9, 17/20349-9, 18/11069-5, 15/50333-1, 17/12622-7)