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Exploring Advantages/Disadvantages and Improvements in Overcoming Gene Delivery Barriers of Amino Acid Modified Trimethylated Chitosan

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Abstract

Purpose

Present study aimed at exploring advantages/disadvantages of amino acid modified trimethylated chitosan in conquering multiple gene delivery obstacles and thus providing comprehensive understandings for improved transfection efficiency.

Methods

Arginine, cysteine, and histidine modified trimethyl chitosan were synthesized and employed to self-assemble with plasmid DNA (pDNA) to form nanocomplexes, namely TRNC, TCNC, and THNC, respectively. They were assessed by structural stability, cellular uptake, endosomal escape, release behavior, nuclear localization, and in vitro and in vivo transfection efficiencies. Besides, sodium tripolyphosphate (TPP) was added into TRNC to compromise certain disadvantageous attributes for pDNA delivery.

Results

Optimal endosomal escape ability failed to bring in satisfactory transfection efficiency of THNC due to drawbacks in structural stability, cellular uptake, pDNA liberation, and nuclear distribution. TCNC evoked the most potent gene expression owing to multiple advantages including sufficient stability, preferable uptake, efficient pDNA release, and high nucleic accumulation. Undesirable stability and insufficient pDNA release adversely affected TRNC-mediated gene transfer. However, incorporation of TPP could improve such disadvantages and consequently resulted in enhanced transfection efficiencies.

Conclusions

Coordination of multiple contributing effects to conquer all delivery obstacles was necessitated for improved transfection efficiency, which would provide insights into rational design of gene delivery vehicles.

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Abbreviations

ANOVA:

Analysis of variance

CLSM:

Confocal laser scanning microscopy

CTB:

Cholera toxin subunit B

DLS:

Dynamic light scattering

DMEM:

Dulbecco’s modified Eagle’s medium

EDC:

1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

GSH:

Glutathione

NC:

Nanocomplexes

NHS:

N-hydroxysuccinimide

pDNA:

Plasmid DNA

SEM:

Scanning electron microscopy

TR:

TMC-Arginine

TRNC:

TR/pDNA nanocomplexes

TRNC2:

TR/TPP/pDNA nanocomplexes

TC:

TMC-Cysteine

TCNC:

TC/pDNA nanocomplexes

TH:

TMC-Histidine

THNC:

TH/pDNA nanocmplexes

TMC:

Trimethyl chitosan

TNC:

TMC/pDNA nanocomplexes

TPP:

Sodium tripolyphosphate

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ACKNOWLEDGMENTS AND DISCLOSURES

The authors would like to thank for the financial support from the National Natural Science Foundation of China (No. 81172995 and No. 81273460).

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Authors and Affiliations

  1. State Key Laboratory of Genetic Engineering Department of Pharmaceutical Sciences, School of Life Sciences, Fudan University, Shanghai, 200433, China

    Hao Zheng, Cui Tang & Chunhua Yin

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  1. Hao Zheng
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  2. Cui Tang
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  3. Chunhua Yin
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Correspondence to Cui Tang.

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Zheng, H., Tang, C. & Yin, C. Exploring Advantages/Disadvantages and Improvements in Overcoming Gene Delivery Barriers of Amino Acid Modified Trimethylated Chitosan. Pharm Res 32, 2038–2050 (2015). https://doi.org/10.1007/s11095-014-1597-7

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  • DOI: https://doi.org/10.1007/s11095-014-1597-7

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