Alkylated Sulfonium Modification of Low Molecular Weight Polyethylenimine to Form Lipopolymers as Gene Vectors

Hydrophobic modification of low molecular weight polyethylenimine (PEI) is an efficient method to form ideal gene-transfer carriers. Sulfonium-a combination of three different functional groups, was conjugated onto PEI 1.8k at a conjugation ratio of 1:0.1 to form a series of sulfonium PEI (SPs). These SPs were hydrophobically modified and characterized by Fourier transform infrared and HNMR. DNA-condensing abilities of SPs were tested with gel retardation experiment, and their cytotoxicity was evaluated via the MTT assay. The particle size and zeta potential of SP/DNA nanoparticles were measured and evaluated for cellular uptake and transfection ability on HepG2 cell line. The results showed that the sulfonium moiety was attached to PEI 1.8k with a high yield at a conjugation ratio of 1:0.1. SPs containing longer alkyl chains condensed DNA completely at an SP/DNA weight ratio of 2:1. The formed nanoparticle size was in the range of 168-265 nm, and the zeta potential was +16-45 mV. The IC₅₀ values of SPs were 6.5-43.2 μg/mL. The cytotoxicity of SPs increased as the hydrophobic chain got longer. SP/DNA showed much stronger cellular uptakes than PEI 25k; however, pure SPs presented almost no gene transfection on cells. Heparin release experiment showed that SP's strong binding of DNA resulted in low release of DNA and thus hindered the gene transfection process. By mixing SP with PEI 1.8k, the mixture presented adjustable DNA binding and releasing. The mixture formed by 67% SP and 33% PEI 1.8k showed strong gene transfection. In conclusion, sulfonium is an effective linkage to carry hydrophobic groups to adjust cell compatibilities and gene transfection capabilities of PEI.