Sustained Transgene Expression Via Substrate-Mediated Gene Transfer Results from Multiple Transfection Events

摘要

Sustained delivery of therapeutic genes in vitro and in vivo has a wide range of applications in studying biology and in developing therapies to treat disease. Non-viral vectors such as cationic polymers still present promising approaches; however, bolus transfection methods with polyethyleneimine (PEI)-based DNA polyplexes suffer from considerable levels of cytotoxicity and short-lived transgene expression levels. Here, we designed and characterized a hyaluronic acid (HA)-based porous hydrogel system for non-viral gene delivery by loading with surface-associated DNA polyplexes (Figure 1A). With this, we observed enhanced and sustained transgene expression over 30 days of cell culture, with better cell viability, both marked improvements over comparable bolus transfection techniques. Finally, we investigated mechanisms thought to be responsible for the sustained expression profile.