Stabilization of a hyaluronate-associated gene delivery system using calcium ions

A "DPH" ternary complex consisting of plasmid DNA (pDNA), intracellularly degradable polyethyleneimine, and hyaluronic acid (HA) is a promising non-viral gene carrier with low toxicity and good gene transfection efficiency. HA plays a key role in providing an optimal balance between DNA protection and release, but it causes aggregation due to the entanglement of HA chains of neighbouring DPH particles. Here we report that the addition of an optimal level of Ca²⁺ successfully prevents particle aggregation and maintains a relatively small size. The Ca-stabilized DPH is comparable to DPH in cytotoxicity and gene transfection efficiency. MW monitoring and conductometric titration suggest that such size stabilization effect is partly mediated by the complexation between HA and Ca²⁺, which enables intra- and intermolecular interactions of HAs.