Spectrophotometer with UV-RAMAN Optical Fiber - tematic project 2019/08568-2

• Grant number: 21/03697-9
• Support Opportunities: Multi-user Equipment Program
• Start date: July 01, 2021
• End date: June 30, 2028
• Field of knowledge: Biological Sciences - Biophysics - Biophysics of Processes and Systems
• Principal Investigator: Pietro Ciancaglini
• Grantee: Pietro Ciancaglini
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 19/08568-2 - Investigation of the extracellular vesicles (VEs) role in the initiation, propagation, regeneration, and modeling of biological mineralization, AP.TEM
• EMU web page: Página do Equipamento Multiusuário não informada
• Type of equipment: Caracterização de Materiais - Espectroscopia - Óptica (UV-Visível); Caracterização de Materiais - Espectroscopia - Raman
• Manufacturer: Fabricante não informado
• Model: Modelo não informado

Abstract

Extracellular vesicles (EVs) has been related to a wide kind of function like developments in immunology, angiogenesis, and stem-cells biology, as well as several pathologies. EVs are bioactive organelles that can carry genetic information, lipids, proteins, and nucleic acids between cells, inducing many molecular functions associated with signalization and regulation of gene expression in the target cells. There is a special group of EVs called Matrix Vesicles (MVs). These vesicles are protagonists in one of the most accepted theories regarding the first hydroxyapatite crystals formation and mineral propagation during the bone formation in vertebrates. However, there are still gaps in these theories that need to be investigated, for example: What is the effective role of vesicles in the process? What is the role of proteins/enzymes present in the MVs? Which interactions between collagen and proteins/enzymes present in the MVs can be responsible for the nucleation and consequently propagation of mineralization? How these interactions regulate the mineralization process mediated by MVs? How pH and phosphosubstrates influence in the formation and propagation of hydroxyapatite crystals mediated by MVs? Through such questions, the project focuses on EVs obtention with enough quality and quantity to understand the interactions source between native vesicles and osteogenic proteins. Proteoliposomes and monolayers of Langmuir as model of membrane enriched with the main proteins of the mineralization process will be used to determine in detail the mechanism of apatite formation and proteins function during the process. The project is innovative and interdisciplinary having the proponent and research collaborators from biochemistry, physical chemistry, pharmacy and odontology areas. Thus, this project will comprehend several methodology fields such as the state of biophysical art (rheology, fluidity), tool methodologies (AFM-TERS, TEM microscopies, BAM, microanalysis of X-ray energy dispersive spectroscopy, RAMAN e SPR), biochemical methodologies (isolations and characterization of native EVs, production of recombinant proteins), cellular biology (primary chondrocytes, osteoblasts and osteoclasts), and functions and applications of EVs in vitro and in vivo using biomimetic proteoliposomes for bone regeneration. Highly resolution details regarding the source of interactions between native EVs or proteoliposomes with main proteins of the biomineralization process can be obtained. The research proposal will provide detailed information about the mineralization ability of EVs, specifying the nucleation ability, propagation, regeneration, and degradation. Another strong feature of the proposal is the development of methodological approaches that can be generalized for EVs investigation. This project becomes very innovative because very few laboratories over the world focus their research on the discovery of the EVs functions. Native and model of vesicles present a great biotechnological potential and can also be used for inhibitors selection in order to prevent the pathological calcification, as well as to provide the bone regeneration/reabsorption. (AU)