Layer-by-Layer Biomimetic Microgels for 3D Cell Culture and Nonviral Gene Delivery

Author(s)

Carvalho, Bruna G. ^[1] ; Vit, Franciele F. ^[1] ; Carvalho, Hernandes F. ^[2] ; Han, Sang W. ^[3] ; de la Torre, Lucimara G. ^[1]

Affiliation

^[1] Univ Campinas UNICAMP, Sch Chem Engn, Dept Mat & Bioproc Engn, BR-13083852 Campinas - Brazil

^[2] Univ Campinas UNICAMP, Inst Biol, Dept Struct & Funct Biol, BR-13083865 Campinas - Brazil

^[3] Fed Univ Sao Paulo UNIFESP, Dept Biophys, BR-04044010 Sao Paulo - Brazil

• Document type: Journal article
• Source: Biomacromolecules; DEC 2021.
• Web of Science Citations: 0

Abstract

Localized release of nucleic acid therapeutics is essential for many biomedical applications, including gene therapy, tissue engineering, and medical implant coatings. We applied the substrate-mediated transfection and layer-by-layer (LbL) technique to achieve an efficient local gene delivery. In the experiments presented herein, we embeded lipoplexes containing plasmid DNA encoding for enhanced green fluorescent protein (pEGFP) within polyelectrolyte alginate-based microgels composed of poly(allylamine hydrochloride) (PAH), chondroitin sulfate (CS), and poly-L-lysine (PLL) with diameters between 70 and 90 mu m. Droplet-based microfluidics was used as the main process to produce the alginate (ALG)-based microgels with discrete size, shape, and low coefficient of variation. The physicochemical and morphological properties of the polyelectrolyte microgels were characterized via optical microscopy, scanning electron microscopy (SEM), and zeta potential analysis. We found that polyelectrolyte microgels provide low cytotoxicity and cell- material interactions (adhesion, spreading, and proliferation). In addition, the microsystem showed the ability to load lipoplexes and a loading efficiency equal to 83%, and it enabled in vitro surface-based transfection of MCF-7 cells. This approach provides a new suitable route for cell adhesion and local gene delivery. (AU)

• FAPESP's process: 18/18523-3 - Polymeric microparticle synthesis via droplet microfluidics for sustained release of non-viral vectors applied to gene therapy
• Grantee: Bruna Gregatti de Carvalho
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 15/20206-8 - Modulation of monocytes, macrophages and pericytes by the colony stimulating factor genes to treat murine limb ischemia
• Grantee: Sang Won Han
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 18/19537-8 - MICROFLUIDICS AS A TECHNOLOGICAL PLATFORM FOR NANO & BIOTECHNOLOGY
• Grantee: Lucimara Gaziola de la Torre
• Support Opportunities: Regular Research Grants
• FAPESP's process: 17/20341-8 - Polymeric microparticle synthesis via droplet microfluidics for sustained release of non-viral vectors applied to gene therapy
• Grantee: Bruna Gregatti de Carvalho
• Support Opportunities: Scholarships in Brazil - Master