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Evaluation of Polymeric Gene Delivery Nanoparticles by Nanoparticle Tracking Analysis and High-throughput Flow Cytometry
Published on: March 1, 2013
Poly(α-glutamic acid) combined with polycation as serum-resistant carriers for gene delivery
Cuifeng Wang1, Min Feng, Jingjing Deng
1School of Pharmaceutical Sciences, Sun Yat-sen University, University Town, Guangzhou 510006, PR China.
International Journal of Pharmaceutics
|August 4, 2010
Summary
Poly(alpha-glutamic acid) (PGA) addition to DNA/poly(ethyleneimine) (PEI) complexes overcomes serum inhibition, maintaining gene delivery efficiency. This terplex system shows enhanced cellular uptake and reduced cytotoxicity, making it promising for in vivo applications.
Area of Science:
- Biomaterials Science
- Gene Therapy
- Polymer Chemistry
Background:
- Cationic polymers are crucial for gene delivery but their efficiency decreases in serum.
- Serum proteins negatively charge and aggregate polyplexes, hindering in vivo applications.
- Poly(alpha-glutamic acid) (PGA) can shield polyplexes from serum proteins.
Purpose of the Study:
- To evaluate the serum-resistant gene delivery potential of DNA/PEI/PGA terplexes.
- To compare the transfection efficiency of terplexes with traditional PEI and Lipofectamine 2000 in serum-containing media.
- To assess the cytotoxicity and cellular uptake of the novel terplex system.
Main Methods:
- Formation and characterization of DNA/PEI/PGA terplexes.
- In vitro transfection efficiency assays in serum-free and serum-containing media.
- Cytotoxicity assays at varying C/N ratios.
- Confocal laser scanning microscopy (CLSM) for cellular uptake studies.
Main Results:
- DNA/PEI/PGA terplexes formed stable complexes with near-neutral surface charge.
- PEI/PGA carriers exhibited no cytotoxicity at C/N ratios > 0.3.
- Terplexes maintained high transfection efficiency in serum, unlike PEI 25 kDa and Lipofectamine 2000.
- CLSM confirmed enhanced cellular import of pDNA delivered by terplexes in serum.
Conclusions:
- PGA effectively overcomes the inhibitory effect of serum on polycation-based gene delivery.
- DNA/PEI/PGA terplexes offer a promising, serum-resistant platform for gene therapy.
- PGA is a versatile reagent for enhancing the in vivo applicability of gene delivery vectors.
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