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Polymeric microparticle synthesis via droplet microfluidics for sustained release of non-viral vectors applied to gene therapy

Grant number: 17/20341-8
Support type:Scholarships in Brazil - Master
Effective date (Start): March 01, 2018
Effective date (End): November 30, 2018
Field of knowledge:Engineering - Chemical Engineering
Cooperation agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:Lucimara Gaziola de la Torre
Grantee:Bruna Gregatti de Carvalho
Home Institution: Faculdade de Engenharia Química (FEQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
Associated research grant:15/20206-8 - Modulation of monocytes, macrophages and pericytes by the colony stimulating factor genes to treat murine limb ischemia, AP.TEM

Abstract

This research project aims to develop alginate/fibroin and alginate/chondroitin sulphate polymer blends to synthesize hybrid hydrogel microparticles. These hybrid microparticles will be used as micro-carriers in gene therapy for the sustained release of non-viral vectors. Gene therapy requires the transfer of genetic materials by transfection or transduction of cells; and thus needs specific delivery systems, such as viral or non-viral vectors. In this project, DNA will be incorporated into cationic liposomes (LCs) before microencapsulation. Liposomes are auto-aggregating amphiphilic lipid vesicles, which resemble cellular membranes. LCs will be produced in microfluidic hydrodynamic flow focusing and complexed with plasmid DNA (pDNA) (LC- pDNA) that encodes the Green Fluorescent Protein (GFP) (model). The micro-carriers for the LC-pDNA encapsulation will be produced by droplet microfluidics. Droplet microfluidics is a promising tool for synthesis of such structures; wherein it is possible to control size, polydispersity, shape, and internal structure. First, in this project, hybrid microparticles will be synthesized in microchannels, which will be compared with a conventional bulk process. Secondly, physical chemistry and mechanical properties of the microparticles will be characterized. Finally, these microparticles will be used as micro-carriers for transfection mammalian cells and sustained release of LCs. Transfection efficiency will be determined by measuring GFP expression level using fluorescence microscopy, and compared with the conventional methods. At the end of this project, it is expected that the work will contribute to the field of droplet-microfluidics, microencapsulation, sustained release, and gene therapy. (AU)