In vitro effect of caffeine in the metabolism and gene expression of osteoblastic cells from bone marrow of ovariectomized rats

• Grant number: 14/19679-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: November 01, 2014
• End date: March 31, 2017
• Field of knowledge: Biological Sciences - Morphology - Cytology and Cell Biology
• Principal Investigator: Karina Fittipaldi Bombonato Prado
• Grantee: Carolina Alves Freiria de Oliveira
• Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Bone remodeling helps bone tissue renovation as well as storage of essential molecules like calcium and magnesium. This process involves proteins, hormones and other substances that are able to activate the well functioning of osteoblasts and osteoclasts. Disturbances in this process may cause osteoporosis, a common disease in a population that nowadays presents higher longevity, inducing loss of bone density, mainly in women with reduction of hormonal load. Several nutritional factors may influence bone remodeling, and among them the habit of drinking coffee, very present in our everyday routine. The caffeine found in this beverage is widely present in several other drinking products and it has great economical importance. The goal of this investigation is to evaluate the in vitro effect of caffeine in the metabolism of osteoblastic cells from bone marrow in an experimental model of osteoporosis. Female wistar rats will be submitted to bilateral ovariectomy in agreement to protocols well established in the literature. After 60 days of the surgical process, the rats will be sacrificed and femurs removed for bone marrow cells isolation in culture flasks with adequate culture medium. After confluence, cells will be seeded in 24-well plates and divided into four experimental groups: 1) control/C (no induction of osteoporosis and no addition of caffeine in media); 2) control/Cc (no induction of osteoporosis and addition of 0,01 mM of caffeine in media); 3) ovariectomized/Ovx (no addition of caffeine in media); 4) ovariectomized/Ovxc (addition of 0,01 mM of caffeine in media). After the experimental periods, the following parameters will be analyzed: cell proliferation and viability, total protein content, alkaline phosphatase activity, detection and quantification of mineralized nodules as well as quantitative expression of genes associated to bone remodeling by means of Real-Time PCR. Data obtained will be analyzed by statistical softwares for significance of pd0.05, with the goal of finding statistical data that may reveal the effects of caffeine in osteoblast metabolism.