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Evaluation of osteogenesis and transcriptional profiles (mRNA and microRNAs) of osteoblastic cells from calvaria and bone marrow of female rats submitted to osteoporosis


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)

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(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)
SEMEGHINI, MAYARA SGARBI; DE AZEVEDO, FERNANDA GRILO; FERNANDES, ROGER RODRIGO; ASSIS, AMANDA FREIRE; DERNOWSEK, JANAINA ANDREA; ROSA, ADALBERTO LUIZ; SIESSERE, SELMA; PASSOS, GERALDO ALEIXO; BOMBONATO-PRADO, KARINA FITTIPALDI. Menopause transition promotes distinct modulation of mRNAs and miRNAs expression in calvaria and bone marrow osteoblastic cells. Cell Biology International, v. 42, n. 1, p. 12-24, . (12/17747-9, 12/15889-0)

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