Folic Acid and Glucose Functionalized PEG-PEI-Based Multifunctional Nanocapsules for Targeted Gene Delivery to Hs-578T Cancer Cells

Document Type : Original Article

Authors

Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran

Abstract

Introduction: Targeted gene delivery can overcome limitations of conventional chemotherapy. In this study, PEI/PEG-based multifunctional nanocapsules with dual targeting were designed by simultaneous modification with folic acid and glucose to enhance gene transfer efficiency into Hs-578T breast cancer cells.
Materials and Methods: The synthesized nanocapsules were characterized using 1H-NMR, FTIR, and TGA. Particle size, morphology, and zeta potential were evaluated by DLS and TEM. Cytotoxicity, DNA binding and protection, and transfection efficiency of the target sequence (pEGFP-N1) were assessed using MTT assay, gel electrophoresis, flow cytometry, and fluorescence microscopy.
Results: DLS analysis showed that PPF, PPG, and PPGF nanocapsules had average sizes ranging from 174 to 301 nm, with folic acid–modified nanocapsules exhibiting larger sizes than glucose-modified ones. Increasing the nanoparticle/DNA ratio reduced particle size and shifted the surface charge from −7.4 to +6.2 mV. DNA release studies demonstrated significantly higher release from PPG, PPF, and PPGF nanocapsules at pH=5.5 compared to pH=7.4, reaching 64.96% in the presence of dextran (100 µg.mL-1). Moreover, folic acid modification enhanced DNA release compared to glucose. MTT results indicated that PEI/DNA complexes induced over 50% cytotoxicity at concentrations above 30 µg/mL, whereas modified nanocapsules showed improved biocompatibility. Gel electrophoresis, flow cytometry, and fluorescence microscopy confirmed effective DNA binding, enzymatic protection, and efficient gene delivery, with dual-ligand modification achieving the highest transfection efficiency.
Conclusion: Dual targeting with folic acid and glucose significantly enhances targeted gene delivery efficiency.

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