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Expression of microRNA-140 inside extracellular vesicles secreted by equine synovial membrane-derived mesenchymal stem cells

Grant number: 19/05558-6
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): November 01, 2019
Effective date (End): February 28, 2021
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Clinics and Surgery
Principal Investigator:Ana Liz Garcia Alves
Grantee:João Pedro Hübbe Pfeifer
Home Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Currently, the application of innovative biotechnologies can be observed in the seeking for an understanding of the developmental processes of musculoskeletal system diseases. These processes generate public health issues in humans and reduce life expectancy in humans and animals. In scientific research that use equine experimental model in orthopaedic diseases, such as Osteoarthritis, the results obtained from these findings could be extrapolated translationally for Human Medicine, since horses develop naturally Osteoarthritis, as well as humans. Aiming to use biotechnologies applicable to both species, it was decided to develop this project in order to evaluate biological factors such as microRNAs involved in the development of equine Osteoarthritis associated with therapeutic mechanisms in the area of regenerative therapy. The objective of this study is to characterize Extracellular Vesicles (EVs) secreted by Synovial Membrane-Derived Mesenchymal Stem Cells (SMMSC) and to evaluate the expression of intravesicular microRNA-140 in two cell culture models, monolayer cultured cells and in alginate hydrogel microcapsules. As well as, to compare the secretion of EVs and the expression of microRNA when challenged in the Osteoarthritis environment in vitro. It has been hypothesized that SMMSCs are capable of secreting EVs expressing microRNA-140 due to the proximity between SMMSCs and chondrocytes in the process of cell development and differentiation. In addition the expression of microRNA-140 is favored by the three-dimensional environment promoted by microcapsules in which the SMMSCs will be cultured. For this investigation, a total of 96 cultures (N=96) will be performed and bioengineering techniques and analytical biotechniques will be used, such as cells microencapsulation, quantification of eca-mir-150-5p expression (equine microRNA-140) by qPCR, global analysis of microRNAs expressed in EVs by transcriptome (NGS), analysis of concentration and size of EVs by Nanosight and transmission electronic microscopy. In order to carry out this project, we highlight the collaboration of Prof. Willian Zambuzzi (IBB, UNESP), who has experience in microvesicles and international research collaboration in this field. (AU)