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Endothelial tissue spheroids: acquisition, charactherization and their osteopromotor potential

Grant number: 16/20505-8
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): July 01, 2017
Effective date (End): April 30, 2019
Field of knowledge:Biological Sciences - Biochemistry - Chemistry of Macromolecules
Acordo de Cooperação: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:Willian Fernando Zambuzzi
Grantee:Geórgia da Silva Feltran
Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
Associated research grant:14/22689-3 - Microvesicle/proteins-mediated paracrine signaling among bone and endothelial cells during bone development and regeneration, AP.JP
Associated scholarship(s):18/18785-8 - Endothelial-induced osteogenic phenotype: lessons from ultra-structural analysis of equivalent tissue, BE.EP.MS


With the progressive increase in population life expectancy, bone defects have become a public health problem. In turn, the skeletal system houses a set of cells, in a hierarchical manner, maintain bone formation throughout life, and its compromised regenerative capacity during the aging process. Despite its medical, social and economic importance, little progress has been made regarding the bioassay methodologies able to rank new therapies. We know that the development and bone regeneration is a complex event orchestrated by paracrine mechanisms of intercellular signaling showing osteogenesis is coupled to angiogenesis. Since 2015 and with support from FAPESP, we have sought to understand the crosstalk mechanisms between endothelial cells and osteoprogenitor cells in order to establish a map of the key molecules involved and transduction mechanisms triggered signals. In this context, we have some students already working on this theme, but exploring in in vitro models traditional. The objective of this project will be to establish spheroids (equivalent tissue to the original) of endothelial tissue through the association of endothelial vessel cells (HUVEC) and artery (HCAEC) with smooth muscle cells (AoSMC) and explore methodologies to characterize it. Three primary cell types have been acquired from the LONZA and are in cultivation in our cell culture unit of LABIO (IBB-UNESP). After these initial steps, already ongoing by the candidate, we will do a proteomic analysis of secretome from these spheroids, and make use of this medium conditioned by spheroids to treat osteogenic cells (h.FOB1.19) in order to know its impact on osteogenesis routes; Here we will analysis of alkaline phosphatase in vitro mineralization, as well as qPCR and Western blotting. It is noteworthy that this request is covered by our JP project, process 2014 / 22689-3, obtained since December 2015, whose main objective is to map the repertoire of molecules secreted by osteoblasts and endothelial cells, and the paracrine involvement between them. (AU)

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Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
FELTRAN, Geórgia da Silva. Generation and characterization of endothelial equivalent tissue and its osteopromotor potential. 2019. Master's Dissertation - Universidade Estadual Paulista (Unesp). Instituto de Biociências. Botucatu Botucatu.

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