Pentablock copolymers of pluronic F127 and modified poly(2-dimethyl amino)ethyl methacrylate for internalization mechanism and gene transfection studies

Abstract

Cationic polymers are one of the major nonviral gene delivery vectors investigated in the past decade. In this study, we synthesized several cationic copolymers using atom transfer radical polymerization (ATRP) for gene delivery vectors: pluronic F127-poly(dimethylaminoethyl methacrylate) (PF127-pDMAEMA), pluronic F127-poly (dimethylaminoethyl methacrylatetert-butyl acrylate) (PF127-p(DMAEMA-tBA)), and pluronic F127-poly(dimethylaminoethyl methacrylate-acrylic acid) (PF127-p(DMAEMA-AA)). The copolymers showed high buffering capacity and efficiently complexed with plasmid deoxyribonucleic acid (pDNA) to form nanoparticles 80-180 nm in diameter and with positive zeta potentials. In the absence of 10% fetal bovine serum, PF127-p(DMAEMA-AA) showed the highest gene expression and the lowest cytotoxicity in 293T cells. After acrylic acid groups had been linked with a fluorescent dye, the confocal laser scanning microscopic image showed that PF127-p(DMAEMA-AA)/pDNA could efficiently enter the cells. Both clathrin-mediated and caveolae-mediated endocytosis mechanisms were involved. Our results showed that PF127-p(DMAEMA-AA) has great potential to be a gene delivery vector.

Surface Charges and Shell Crosslinks Each Play Significant Roles in Mediating Degradation, Biofouling, Cytotoxicity and Immunotoxicity for Polyphosphoester-based Nanoparticles

“…The PPE nanoparticles developed in the current study can be rapidly synthesized and provide versatile nanostructures (anionic, neutral, cationic or zwitterionic), with control over their surface chemistries to accommodate and deliver anionic, cationic, and/or hydrophobic guest (macro)molecules. In addition, the cytotoxicity data reveal that PPE-based nanoparticulates may have a remarkable safety profile, as compared to commercially-available vectors, and degradable/non-degradable nanoparticles developed previously by our group 39 40 , and by others 41 42 .…”

Nano-On-Nano: Responsive Nanosubstrate-Mediated Liposome Delivery with High Cellular Uptake Efficiency

“…HEK-293T were seeded in 6-well plates at a density of 1 × 10 5 cells per well and incubated for 21 h with DMEM, 5% CO 2 at 37 °C. The culture media was removed after incubation and cells were once again incubated with 1 mL of endocytic inhibitors as chlorpromazine (10 μM), 30 sodium azide (0.1% w/v in medium), 31 genistein (200 μM), 32 wortmannin (50 nM), 32 and nocodazole (33 μM) 33 for 1 h at 37 °C. Further, cells were washed with PBS, trypsinized, centrifuged, and injected into liposomes coated on the nanosubstrate with a flow rate of 0.2 mL/min and seeded for 10 min thermal stimulation cycles were applied to detached calcein-loaded liposomes for cellular uptake.…”