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The role of bone morphogenetic protein 2 (BMP-2) and of enamel matrix derivative (EMD) in the osteogenic and dentinogenic differentiation


The use of extra-cellular matrix proteins has been suggested as an alternative for tissue repair, mainly in mineralized tissues. Moreover, since their action is still unclear, many studies are necessary to understand the intracellular mechanisms involved in the repair process, as well as the use of these processes towards tissue engineering. In this context, not much is known regarding the signaling pathways activated during cell differentiation, and regarding the transcription factors involved. The purpose of this study will be to observe the effects of BMP-2 and EMD on cells from the dental pulp (fibroblasts and stem cells) and cells from bone (osteoblasts), regarding the activation of signaling pathways, BMP receptors involved and activated transcription factors, specifically Dlx3, Dlx5, Msx2 and Runx2. The differentiation and mineralization processes will be confirmed by PCR reactions (Osteocalcin - OCN, Osteopontin - OPN, Bone sialoprotein - BSP, Dentin matrix protein 1 - DMP1 and dentin sialophosphoprotein - DSPP), as well as by enzymatic activity for alkaline phosphatase and mineralized nodule formation (Alizarin red staining). The protein expression of BMP receptors (Ia, Ib and II), the activation of the canonical TGF pathway, of the canonical Wnt pathway and the activation of MAPKinase pathway will be analyzed by Western Blotting. Finally, the expression of development genes will be quantified by qPCR. (AU)

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