Polyethyleneimine/DNA polyplexes with reduction-sensitive hyaluronic acid derivatives shielding for targeted gene delivery

The natural anionic polysaccharide hyaluronic acid (HA) was modified by introducing reduction-sensitive disulfide bond between the carboxyl groups and the backbone of HA (HA-SS-COOH). HA-SS-COOH and its corresponding unmodified stable analog HA were used to shield DNA/PEI polyplexes (DP) to form ternary complexes (DPS and DPH complexes). The shielding/deshielding effect was tested along with size, zeta potential, cell viability and transfection. Both DPS and DPH complexes showed increase in size, decrease in zeta potential and low cytotoxicity in physiological conditions due to the anionic shielding. In the reductive environment, only HA-SS-COOH coated ternary complexes (DPS) demonstrated the size increase and recovered high positive zeta potential. DPS complexes showed an up to 14-fold higher transfection than the stable coated one, indicating the efficiency of the reduction-responsive deshielding design. Moreover, the presence of extra free HA inhibited the transfection of DPS on HepG2 and B16F10 cells with HA receptor expression, while displaying no effect on non-targeted NIH3T3 cells. More rapid cellular association of DPS with HepG2 was observed, thus confirming the targeting reservation of disulfide bond modified HA. Intratumoral injection of DPS complexes resulted in much higher accumulation and luciferase expression in the tumor bearing C57BL/6 mice. Both in vitro and in vivo results demonstrated the successful combination of deshielding and target functions in HA derivatives for gene delivery.