A Proof-of-Concept Preparation of Lipid-Plasmid DNA Particles Using Novel Extrusion Based 3D Printing Technology, SMART

Gene therapy is a promising approach with delivery of messenger RNA, small interference RNA, and plasmid DNA to elicit a therapeutic action in vitro using cationic or ionizable lipid nanoparticles. In the present study, a novel extrusion based Sprayed Multi-Adsorbed droplet Reposing Technology (SMART) developed in-house was employed for preparation, characterization, and transfection abilities of the green fluorescence protein (GFP) plasmid DNA in cancer cells in vitro. The lipid composition (ionizable) and plasmid DNA (pDNA, GFP) were mixed in a 1:1 ratio using SMART technology at 1, 5, 8 & 10 N/P ratios. The particles were characterized to determine particle size (DLS), zeta potential and morphology using scanning electron microscopy (SEM). The particle size yielded for all N/P ratios is in the range of 100 nm to 200 nm. The in vitro transfection was carried out in MG63 cells showed optimal formulation N/P 8 with expression of GFP protein. The toxicity study through MTT assay showed N/P 8 with toxicity lower than other groups. The results showed that the processes developed using SMART technology are consistent and can be utilized for commercial applications.