Intercalation of shRNA-plasmid in Mg-Al layered double hydroxide nanoparticles and its cellular internalization for possible treatment of neurodegenerative diseases

摘要

In the present work, nanoconjugates of shRNA-plasmid and a non-viral nanoceramic vector, e.g., Mg-Al layered double hydroxide (Mg-Al LDH), were synthesized and intercalated. Subsequently, these particles with an average size of 40-60 nm, were transfected into mammalian neuroblastoma cells (SH-SY5Y). The as prepared Mg-Al LDH was able to protect the incorporated shRNA-plasmid against a range of pH values, DNasel, endonucleases, and serum components. To test the applicability of the nanoconjugate for future in-vivo studies, serum from three different model experimental animals viz, mouse, rat and guinea pig was used for the serum protection study. Additionally, we showed that prolonged storage at different temperatures does not affect the quality of the nanoconjugate. Using this nanoconjugate to transform cells, a maximum internalization of ~ 26% at 24h was achieved. Lastly, we demonstrated effective and safe delivery of the plasmid by measuring GFP production and shRNA-induced knockdown of TNF alpha.