Fabrication of mRNA encapsulated Lipid nanoparticles usingState-of-the-artSMART-MaGIC Technology and transfectionin vitro

Abstract

AbstractThe messenger ribose nucleic acid (mRNA)in lieuof Corona virus of 2019 (COVID-19) vaccines were effectively delivered through Lipid nanoparticles (LNP) which were proved effective carriers for clinical applications. In the present work, mRNA (erythropoietin (EPO)) encapsulated LNPs were prepared using a next generation state-of-the-art patented Sprayed Multi Absorbed-droplet Reposing Technology (SMART) coupled with Multi-channeled and Guided Inner-Controlling printheads (MaGIC) technologies. The LNP-mRNA were synthesized at different N/P ratios and the particles were characterized for particle size & zeta potential (Zetasizer), encapsulation or complexation (gel retardation assay) and transfection (Fluorescence microscopy) in MG63 sarcoma cellsin vitro. The results showed a narrow distribution of mRNA-lipid particles of 200 nm when fabricated with SMART alone and then the size was reduced to approximately 50 nm with the combination of SMART-MaGIC technologies. The gel retardation assay showed that the N/P>1 showed strong encapsulation of mRNA with lipid. Thein vitroresults showed the toxicity profile of the lipids where N/P ratio of 5 is the optimized with >50% cell viability. The functional LNP-mRNA were prepared and analyzed with SMART-MaGIC technologies which could be a potential new fabrication method of mRNA loaded LNPs.HighlightsThe particle size was reduced to around 50 nm with implementation of SMART-MaGIC.The loading efficiency is 100%.The functionality of the mRNA is unaffected during the preparation process.The transfection facilitated transient expression of the proteinin vitro.The EPO mRNA is more effective than EPO protein to reduce chemo-toxic effectsin vitro.