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Comparative analysis of the effect of osteoporosis on the in vitro behavior of bone marrow and calvaria osteoblastic cells from female ovariectomized rats

Grant number: 12/13330-6
Support type:Scholarships in Brazil - Master
Effective date (Start): November 01, 2012
Effective date (End): February 28, 2014
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Karina Fittipaldi Bombonato Prado
Grantee:Fernanda Grilo de Azevedo Fernandes
Home Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil


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 osteoporotic female rats the behavior of bone marrow mesenchymal cells differentiated in osteoblasts with osteoblasts from calvaria. There will be used 18 wistar female rats divided in control and osteoporotic groups. Osteoporosis will be induced through bilateral ovariectomy and after 150 days of surgery, both groups will be sacrificed to collect the femurs and calvaria 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. Besides, it will be evaluated the expression of proteins associated to osteogenesis (ER-±, ER-², RANKL, RUNX2, ALP, OCN) through immunolocalization as well as quantitative expression of their related genes through PCR Real Time. The data obtained will be analyzed through adequate statistical test through SPSS software 17.0 and the significance will be 5%.