Behavior of endothelial and smooth muscular cells under shear-stress model: a panorama of cellular, ultrastructural and biochemistry aspects

Abstract

Our main proposal has been to develop methodologies capable of unraveling the mechanisms involved in paracrine crosstalk between endothelial cells and osteoprogenitor cells, in order to map the repertory mechanisms involved in this context. Much of these methodologies have been developed by subjecting the endothelial cells to Shear Stress (SS) model and then the conditioned medium from them used for the treatment of undifferentiated cells to know their osteopromotor potential. Although endothelial cells respond well to chemical agonists which stimulate the receptors coupled to G protein, they also respond to mechanical forces produced by the blood flow, synthesizing and secreting chemical factors involved in numerous physiological processes, such as coagulation, fibrinolysis, angiogenesis and maintenance of vascular tone; it has been showed also its intimate relationship with osteogenesis processes as proposed by Kusumbe et al., 2014. In this sense, our proposal is to characterize the cellular and molecular alterations caused by shear stress on endothelial cells (arterial and venous) and smooth muscle cells. Specifically, we will: 1. Analyze proteomic characterization of secretome endothelial and muscle cells; 2. Analyze potential signal transduction pathways involved in rearrangement of actin filaments; 3. Estimate the influence of the model mechanisms remodeling of extracellular matrix by the expression and activity of matrix metalloproteinase (MMP2 and MMP9); 4. To investigate the effect of the model in the morphology and ultrastructures of those cells by electron microscopy and fluorescence; 5. Determination of epigenetic mechanisms by analyzing the levels of 5meC and 5hmeC. A guarantee of success of this project is the investment of $ 1.3 million by FAPESP (JP-2014 / 22689-3) for equipping the LaBIO: bioassays and Dynamics Cell Laboratory, placed in the Department of Chemistry and Biochemistry, under our responsibility. Establishing biochemical events, cell and ultrastructural of endothelial cells (arterial and venous) and smooth muscle cells, subjected or not to the model Shear Stress, is extremely important at this point to establish the repertoire synthesized and secreted proteins due the SS model, adding significant results to our line of research, where new projects are located. Another important point is contributing for formation of human resources, in alliance with the development of the group our research group here at UNESP-Botucatu. (AU)