Synthesis of Mannosylated Polyethylenimine and Its Potential Application as Cell-Targeting Non-Viral Vector for Gene Therapy

Abstract

Mannose polyethylenimine with a molecular weight of 25 k (Man-PEI25k) was synthesized via a phenylisothiocyanate bridge using mannopyranosylphenyl isothiocyanate as a coupling reagent, and characterized by 1 H NMR (nuclear magnetic resonance) and FT-IR (Fourier transform infrared spectroscopy) analysis. Spherical nanoparticles were formed with diameters of 80-250 nm when the copolymer was mixed with DNA at various charge ratios of copolymer/DNA (N/P). Gel electrophoresis demonstrated that the DNA had been condensed and retained by the PEI derivates at low N/P ratios. The Man-PEI25k/DNA complexes were less cytotoxic than the PEI complexes with a molecular weight of 25 k (PEI25k) at the same N/P ratio. Laser scan confocal microscopy and flow cytometry confirmed that the Man-PEI25k/DNA complexes gave higher cell uptake efficiency in (Dendritic cells) DC2.4 cells than HeLa cells. The transfection efficiency of Man-PEI25k was higher than that of PEI25k towards DC2.4 cells. These results indicated that Man-PEI25k OPEN ACCESSPolymers 2014, 6 2574 could be used as a potential DC-targeting non-viral vector for gene therapy.

Biopolymeric Materials Used as Nonviral Vectors: A Review

“…Table 2 presents the main factors evaluated in the articles about to the efficiency of gene transfection and bacterial transformation by different biopolymers and polysaccharides derivates: chitosan [19], dextran [20,21], quaternized cellulose [22] and quaternized chitosan [23]. Other types of substances were used as nonviral vectors such as: plasmid DNA nanostructures with ionic liquid [11], pegylated polyethyleneimine nanoparticles conjugated with folate and galactose [24], cationic liposomes modified with polyallylamine [25], biodegradable polylactic acid-polyethylene glycol-poly (L-lysine) copolymer [26], crotamine [27], poly (oligo-D-arginine) [28], poly (lactic acid-co-glycolic acid) scaffolds [29], polyethyleneimine/pDNA nanocomplexes with anionic hyaluronic acid [30], fourth generation cationic phosphorus (P4) containing dendrimers [31], and monosylated polyethyleneimine [32]. All the materials tested on these studies showed low cytotoxicity and enhanced significantly the efficiency (p < 0.05).…”

“…Polyethyleneimine (PEI) is one of the most successful and efficient nonviral gene transfer systems that have been reported to date. In four of the fifteen articles [19,24,30,32], PEI was used as the main polymer modified and used as a vector of gene transport.…”

Application of Mono‐ and Disaccharides in Drug Targeting and Efficacy

“…Also, due to the enhanced antigen uptake and presentation by APC, mannosylated antigen has been shown to be a potential approach to potentiate antigen immunogenicity. Mannosylated PEI is an alternative targeting approach to tumor‐associated macrophages (TAMs) and DC, and acts as a cell‐targeting non‐viral vector for gene therapy . Mannosylated dendrimer ovalbumin (MDO) potently elicited OVA‐specific T‐cell responses in vitro owing to its strong avidity for DC, indicating that it is a potent immune response inducer.…”

“…[99] MRs bind with high affinity to infectious agents containing terminal mannose residues,t riggering the transport of the stimulants into endocytic pathways, finally resulting in major histocompatibility complex( MHC) presentation and subsequentT -cell activa-tion. [100] The incorporation of mannose into variousn on-viral delivery carriers such as polyethyleneimine (PEI), [101][102][103][104][105][106][107][108] lipids, [31,[109][110][111] gelatin, [112] and catalase [113] has been demonstrated to produce an APCt argeting ability and high transfection efficiency.A lso, due to the enhanceda ntigen uptake and presentation by APC, mannosylated antigen has been shown to be apotential approachtopotentiate antigen immunogenicity. Mannosylated PEI is an alternative targetinga pproacht o tumor-associated macrophages (TAMs) and DC,a nd acts as a cell-targeting non-viral vector for gene therapy.…”

“…Mannosylated PEI is an alternative targetinga pproacht o tumor-associated macrophages (TAMs) and DC,a nd acts as a cell-targeting non-viral vector for gene therapy. [101][102][103][104][105][106][107][108] Mannosylated dendrimer ovalbumin (MDO) potently elicited OVA-specific T-cell responsesi nvitro owing to its strong avidity for DC, indicating that it is ap otent immune response inducer.F urthermore, the immunogenicityo fMDOi sa scribed not only to the intensification of antigen presentation, but also to the inductiono fD Cm aturation;t herefore, MDO is also considered to be ap otential vaccinec arrier. [114] Mannosylated dextran nanoparticles are another case of successful vaccine carriers.…”