Evaluation of transfection with gemini nanoparticles for retinal neuroprotection in a dynamic microfluidic cell culture system

Abstract

<img src=” https://s3.amazonaws.com/production.scholastica/article/94464/large/prnano_1252024ga.jpg?1709170827”> Glaucoma is a neurodegenerative disease of the retina and optic nerve with elevated intraocular pressure as one of the main contributing factors. The traditional in vitro methods to evaluate new gene delivery systems for neuroprotective treatments of glaucoma require testing in cell monolayers under static conditions. Therefore, a microfluidic technology that provides a dynamic simulation of cell exposure to nanoparticles (NPs) could be a useful model for rapid screening. In this study, CellASIC ONIX2 microfluidic platform was used to evaluate and optimize culture and transfection conditions to assess cell viability and transfection efficiency (TE) of gemini NPs carrying GFP-encoding plasmid in A7 astrocytes, one of the targeted retinal cells for glaucoma treatment. The average TE (inlet-middle-outlet of culture chambers) by gemini nanoplexes, 18-7NH-18 NPXs and 18-7Np3-18 NPXs was > 85% and > 25%, respectively, after 5 h of perfusion with a combined average FI (caFI) of about 600 and < 100, respectively, whereas TE after Lipofectamine 3000 was > 70% and caFI of < 100. The results have shown that comparatively to 2D monolayers, the microfluidics technique provides better exposure of cells to NPs, suitable for kinetic mapping of gene expression, and a useful tool for cell growth and live evaluation of gene delivery system interaction with cells.