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

• Grant number: 17/20341-8
• Support Opportunities: Scholarships in Brazil - Master
• Start date: March 01, 2018
• End date: November 30, 2018
• Field of knowledge: Engineering - Chemical Engineering
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Lucimara Gaziola de la Torre
• Grantee: Bruna Gregatti de Carvalho
• Host 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)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

CARVALHO, BRUNA G.; VIT, FRANCIELE F.; CARVALHO, HERNANDES F.; HAN, SANG W.; DE LA TORRE, LUCIMARA G.. Layer-by-Layer Biomimetic Microgels for 3D Cell Culture and Nonviral Gene Delivery. Biomacromolecules, DEC 2021. (18/18523-3, 15/20206-8, 18/19537-8, 17/20341-8)