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In vitro evaluation of functional activity of osteoblastic cells MCT3-E1 cultured with conditioned medium from raw 264.7 cell line macrophages

Grant number: 18/23587-0
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): February 01, 2019
Effective date (End): April 30, 2020
Field of knowledge:Biological Sciences - Morphology - Histology
Principal researcher:Karina Fittipaldi Bombonato Prado
Grantee:Sayuri Poli Suguimoto
Home Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil


Osteogenesis is achieved not only by osteoblastic cells but also as a result from a cooperation with several systems such as imune system. Macrophages may secrete several signaling molecules wich may affect in a good or bad way the osteogenic capacity of osteoblasts. Thus, the objective of this project will be to evaluate the influence of a macrophage conditioned medium in the culture of MC3T3-E1 osteoblastic cells. After the storage of conditioned medium, osteoblastic cells will be cultured in flasks in osteogenic medium until subconfluence and seeded at a concentration of 2 x 104 cells per well in culture plates (n = 5). Experimental groups will be divided in control cells (C) and treated with 50% of conditioned medium (MC). After the experimental period, the following parameters will be evaluated: cell proliferation, total protein contente, ALP activity and its in situ detection, and quantification of mineralized nodules. The results will be analyzed by statistical programs consistent with significance p d 0,05, with the goal of finding statistically significant the effects conditioned medium in the functional activity of osteoblasts.

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