Redox-Sensitive Gelatin/Silica-Aptamer Nanogels for Targeted siRNA Delivery.

RNA interference (RNAi) has potential advantages over other gene therapy approaches due to its high specificity and the ability to inhibit target gene expression. However, the stability and tissue-specific delivery of siRNA remain as the biggest obstacles for RNAi therapeutics. Here, we developed such a system by conjugating gelatin-based nanogels with the nucleolin-targeted AS1411 aptamer and deoxynucleotide-substituted siRNA together (Apt-GS/siRNA) via a disulfide linker to achieve transient docking of siRNA. These Apt-GS/siRNA nanogels demonstrated favorable release of siRNA under reducing conditions owing to disulfide cleavage. Furthermore, this smart system could electively release siRNA into the cytosol in nucleolin-positive cells (A549) by a glutathione-triggered disassembly and subsequently efficient RNAi for luciferase. Besides, disulfide-equipped Apt-GS nanogels showed good biocompatibility in vitro. Taken together, this redox-responsive, tumor-targeting smart nanogels display great potential in exploiting functionalized siRNA delivery and tumor therapy.

Enhanced activity and stability of papain by covalent immobilization on porous magnetic nanoparticles.

Papain enzyme was successfully immobilized by covalent bonding onto biocompatible Fe3O4/SF nanoparticles, which were prepared with the soft template of silk fibroin (SF). The optimized immobilization condition is pH6.0, hydrolysis time of 60min, and an enzyme/support ratio of 10.0mg/g. Compared with free papain, the immobilized papain exhibits a high effective activity, broader working pH and temperature. This immobilized papain can be separated from the solution by the external magnetic field for cyclic utilization, and 70% of initial activity was retained after eight consecutive operations while completely loss of proteolytic activity for the free papain. Furthermore, the immobilized papain maintained 85% of their initial activity after being stored for 28days. Kinetic parameters, maximum reaction rate (Vmax) and Michaelis constant (Km) of immobilized papain, were determined as 4.95mg/l·min and 0.23mg/ml, larger than its free counterpart. All the results above indicated that the immobilized papain onto magnetic Fe3O4/SF nanoparticles would have potential industrial and medical applications.