Large scale gene expression evaluation of osteoblastic cells from calvaria and bon...
| Grant number: | 12/17747-9 |
| Support type: | Regular Research Grants |
| Duration: | January 01, 2013 - December 31, 2014 |
| Field of knowledge: | Biological Sciences - Morphology - Cytology and Cell Biology |
| Principal Investigator: | Karina Fittipaldi Bombonato Prado |
| Grantee: | Karina Fittipaldi Bombonato Prado |
| Home Institution: | Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil |
| Assoc. researchers: | Adalberto Luiz Rosa ; Geraldo Aleixo da Silva Passos Júnior ; Márcio Mateus Beloti ; Paulo Tambasco de Oliveira ; Selma Siéssere |
Abstract
Mesenchymal stem cells are nowadays considered an important source for tissue replacement. These cells have osteogenic potential and can be used for bone tissue maintenance and repair, being very attractive for tissue engineering. Nevertheless, changes in their behavior might occur in consequence of conditions such as osteoporosis, affecting their ability of auto-renovation and differentiation. Thus, the purpose of this investigation is to compare in ovariectomized female rats the behavior of bone marrow mesenchymal cells differentiated in osteoblastic cells with those from calvaria. There will be used 18 wistar female rats divided in control and ovariectomized groups. Osteoporosis will be induced through bilateral ovariectomy and after 150 days of surgery, both groups will be sacrificed to collect the femurs and calvariae fragments. From these collected materials, there will be isolated cells from bone marrow and calvaria which will be cultivated in flasks with culture medium and posteriorly in cell plates of 24 wells (n=5) to evaluate the following parameters: cell proliferation and viability, total protein content, alkaline phosphatase activity (ALP), detection and quantification of mineralized nodules and expression of proteins associated to osteogenesis through immunolocalization and Real-Time PCR. Besides, it will be performed the evaluation of transcriptional profile (mRNA and microRNAs) and gene network construction. Data obtained from biochemical assays wil be analysed through statistical programs for p <0.05 as well as through specialized bioinformatics programs for transcriptional profile data. (AU)