Studies of metabolic incorporation of azido sugars in glycoconjugates during the differentiation of mesenchymal stem cells of mouse

• Grant number: 12/06684-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2012
• End date: May 31, 2014
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: José César Rosa
• Grantee: Thaiene Aizza
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

The growth, development, and maintenance of bone tissue are highly regulated processes. Several proteins such as hormones, growth factors, and cytokines are actively involved in these processes and exert direct activity on osteoblastic and osteoclastic cells, acting in their differentiation and metabolic activation. The process of bone regeneration is initiated by local stimulating factors such as bone morphogenetic protein (BMP - bone morphogenetic proteins). BMPs are a product of the metabolism of osteoblasts, and odontoblasts of various tumor cells, and are stored as concentrated in the bone, dentin and osteosarcoma, and malignant cells of certain odontogenic tumors, such as identifying fibroids, benign cementoblastoma, dentinoma, odontogenic fibroma, and odontoma. Clarifying the mechanisms that control bone remodeling is an issue very important. In this sense, mesenchymal stem cells have attracted great interest because of their potential involvement in tissue repair. The attainment of functional osteoblasts from mesenchymal stem cells has been used in tissue engineering and cell therapy. Therefore, this proposal is to investigate the effects of metabolic incorporation of azide sugar in various stages of osteoblastic differentiation of mesenchymal stem cells obtained from Wistar rat bone marrow with the aim to obtain knowledge of the cell differentiation and several steps of osteogenesis. In this proposal, we intend to use an experimental strategy called a chemical biorthogonal reporter. This strategy includes the metabolic incorporation of azide sugars acetylated analogs of N-acetyl-galactosamine (GalNac), N-acetylglucosamine (GlcNAc), and N-acetilmanosamina (ManNAc) in mesenchymal stem cells. The azido sugars, acetylated, commercially available N-azideacetyl-galactosamine (Ac4GalNAz), N-acetylated azideacetylglucosamine (Ac4GlcNAz), and N-acetylated azide-acetylmannosamine (Ac4ManNAz) are used by normal biosynthetic routes and can be subsequently detected using specific probes.(AU)