Gene transfer on inorganic/organic hybrid silica nanosheets†
Abstract
Gene delivery is often accomplished by the forward or reverse transfection protocol. In either protocol, a transfection reagent (usually cationic) is added to increase the delivery efficiency. In this study, we employed a series of nanosheet networks to facilitate the delivery of naked plasmid DNA into human mesenchymal stem cells (hMSCs). By adding different chemicals into the reaction mixture for etching the silica glass, we were able to fabricate inorganic/organic hybrid nanosheet networks with different physico-chemical characteristics. We then analyzed the transfection efficiency on different nanosheets and the possible dependence of the transfection efficiency on the physico-chemical parameters of nanosheets. The results showed that all nanosheet networks were noncytotoxic and demonstrated a high cell survival rate (∼90%) after transfection. The transfection efficiency was critically determined by the aspect ratio (height/thickness of the wall) of the nanosheets. The effects of chemistry or other surface properties were not significant. Moreover, the transfection efficiency may be successfully predicted by the initial cell migration rate and the activation of integrin β3 on the nanosheets. Compared to the conventional method, transfection using concurrent cell/plasmid seeding on the nanosheets is not only more effective but also much safer. Future efforts may focus on combining the inorganic/organic hybrid nanosheets with soft substrates for in situ transfection.