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Poly (amino ester) Composed of Poly (ethylene glycol) and Aminosilane Prepared by Combinatorial Chemistry as a Gene Carrier

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Abstract

Purpose

Application of combinatorial chemistry and high throughput screening for the synthesis and evaluation of mini-library of novel biodegradable poly (β-amino ester)s (PAE)s composed of γ-aminopropyl-triethoxysilane (APES) and poly (ethylene glycol) diacrylate (PEGDA) for gene delivery efficiency and safety in 293T and HeLa cells in the presence of and absence of serum.

Materials and methods

PAEs were synthesized at different mole ratios of APES and PEGDA by Michael addition reaction and synthesis was confirmed by 1H nuclear magnetic resonance (1H-NMR). Ninety six ratios of polyplexes were evaluated for luciferase and MTS assay in 293T and HeLa cells in the presence of and absence of serum. Relationship between transfection efficiency and DNA binding ability of PAEs was studied by gel electrophoresis. Particle sizes and molecular weight of selected PAEs were measured by dynamic light scattering and gel permeation chromatography multi-angle light scattering, respectively.

Results

1H-NMR confirmed the synthesis of PAEs. In both cell lines, transfection efficiency and cell viability were increased for PAEs obtained from R106 (0.7:1, APES:PEGDA) to R121 (6:1, APES:PEGDA) with a marginal increase in APES concentration. Transfection pattern was uniform in the absence of and presence of serum. In both cell lines, PAE obtained from R121 demonstrated high transfection efficiency and low cytotoxicity as compared to polyethylenimine (25 KDa) and Lipofectamine. PAE obtained from R121 showed good DNA binding and condensation with average particle sizes of 133 nm.

Conclusion

Addition of PEGDA over APES resulted in a novel PAE which has high safety and transfection efficiency. Transfection and cytotoxicity are very sensitive to monomer ratios and mainly governed by concentration of amine monomer.

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Abbreviations

AEAPMS:

N-β-(aminoethyl)-γ-aminopropyl-trimethoxysilane

APES:

γ-aminopropyl-triethoxysilane

DMEM:

Dulbaco’s modified Eagles media

PAE:

poly (β-amino ester)

PEGDA:

poly (ethylene glycol) diacrylate

PEI 25K:

high molecular weight polyethylenimine (25 KDa)

RLU/mg protein:

Relative light units per milligram of protein

R106 to R121:

reaction codes representing PAEs obtained from different mole ratios of APES and PEGDA

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Acknowledgements

This work is funded by Korea Research Foundation (D00248). We also acknowledge National Instrumental Centre for Environmental Management for measurement of NMR and particle size. Mr. Dhananjay Jere is supported by Korea Research Foundation.

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Authors and Affiliations

  1. School of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, South Korea

    Dhananjay Jere, Mi-Kyong Yoo, Rohidas Arote, Tae-Hee Kim,  Yun-Jaie Choi & Chong-Su Cho

  2. College of Veterinary Sciences, Seoul National University, Seoul, 151-742, South Korea

    Myung-Haing Cho

  3. Department of Polymer Science and Engneering, Sunchon National University, Sunchon, 540-742, South Korea

    Jae-Woon Nah

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  1. Dhananjay Jere
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Correspondence to Chong-Su Cho.

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Jere, D., Yoo, MK., Arote, R. et al. Poly (amino ester) Composed of Poly (ethylene glycol) and Aminosilane Prepared by Combinatorial Chemistry as a Gene Carrier. Pharm Res 25, 875–885 (2008). https://doi.org/10.1007/s11095-007-9448-4

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  • DOI: https://doi.org/10.1007/s11095-007-9448-4

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