Galactosylation of Chitosan-Graft-Spermine as a Gene Carrier for Hepatocyte Targeting In Vitro and In Vivo

Abstract

Polyethyleneimine (PEI) has been described as a highly efficient gene carrier due to its efficient proton sponge effect within endosomes. However, many studies have demonstrated that PEI is toxic and associated with a lack of cell specificity despite high transfection efficiency. In order to minimize the toxicity of PEI, we prepared chitosan-graft-spermine (CHI-g-SPE) in a previous study. CHI-g-SPE showed low toxicity and high transfection efficiency. However, this compound also had limited target cell specificity. In the present study, we synthesized galactosylated CHI-g-SPE (GCS) because this modified GCS could be delivered specifically into the liver due to hepatocyte-specific galactose receptors. The DNA-binding properties of GCS at various copolymer/DNA weight ratios were evaluated by a gel retardation assay. The GCS copolymer exhibited significant DNA-binding ability and efficiently protected DNA from nuclease attack. Using energy-filtered transmission electron microscopy (EF-TEM), we observed dense spherical, nano-sized GCS/DNA complexes with a homogenous distribution. Most importantly, GCS was associated with remarkably low cytotoxicity compared to PEI in HepG2, HeLa, and A549 cells. Moreover, GCS carriers specifically delivered the gene-of-interest into hepatocytes in vitro as well as in vivo. Our results suggest that the novel GCS described here is a safe and highly efficient carrier for hepatocyte-targeted gene delivery.

Codelivery of temozolomide and siRNA with polymeric nanocarrier for effective glioma treatment

“…16 In particular, poly(ethylenimine) (PEI) is acknowledged as the most promising one not only because it readily complexes siRNA but also because it facilitates the lyosomal escape of nanocarriers through proton sponge effect. 17 However, the issue may become much more complicated in designing vectors for the codelivery purpose. A main reason is that two different types of therapeutic agents often possess very different physicochemical properties and thus require different structural materials to accommodate them.…”

PK11195-chitosan-graft-polyethylenimine-modified SPION as a mitochondria-targeting gene carrier

“…Besides, PK-CP-SPION maintained excellent DNA protection ability upon the addition of nucleases ( Figure 3B), which further assured the efficient gene delivery to targeting sites [36]. Physicochemical properties, such as particle size and surface charge could greatly affect the cellular uptake of nanoparticles [37,38], thus the hydrodynamic diameter and zeta potential of PK-CP-SPION/DNA complexes at various weight ratios were investigated. As shown in Figure 3(C), the particle size of the complexes decreased (from 120.1 nm to 49.7 nm) whilst the zeta potential increased (from À25.9 to 32.7 mV) as the weight ratio increased.…”

Glycosylation-mediated targeting of carriers

“…[29][30][31][32][33], poly-glutamic acid (PLGA) [34][35][36], polyethlenimine (PEI) [37][38][39][40][41][42], chitosan [43][44][45][46][47], dendrimer/α-cyclodextrin conjugate [48], and their derivatives [49,50] have been studied for galactosylated macromolecular carriers for hepatocyte-selective targeting. The design of carrier types based on the physicochemical properties and galactose density of the carriers are of significance for targeting efficacy by galactosylated macromolecular carriers.…”