Despite their potential, PEI-based systems still face significant limitations in achieving the high delivery efficiencies required for clinical use . A critical factor influencing efficiency is the formation and stability of polyplexes . Strong binding and efficient nucleic acid release must be carefully balanced because overly stable polyplexes can hinder intracellular release, while weak complexes fail to protect and transport cargo effectively into cells . Previous studies have predominantly focused on optimizing transfection efficiency only by adjusting molecular weight , nitrogen-to-phosphate (N/P) ratios , or complex formation conditions .