Stability, intracellular delivery, and release of siRNA from chitosan nanoparticles using different cross-linkers

Maria Abdul Ghafoor Raja, Haliza Katas, Thum Jing Wen

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Chitosan (CS) nanoparticles have been extensively studied for siRNA delivery; however, their stability and efficacy are highly dependent on the types of cross-linker used. To address this issue, three common cross-linkers; tripolyphosphate (TPP), dextran sulphate (DS) and poly-D-glutamic acid (PGA) were used to prepare siRNA loaded CS-TPP/DS/PGA nanoparticles by ionic gelation method. The resulting nanoparticles were compared with regard to their physicochemical properties including particle size, zeta potential, morphology, binding and encapsulation efficiencies. Among all the formulations prepared with different cross linkers, CS-TPP-siRNA had the smallest particle size (ranged from 127 ± 9.7 to 455 ± 12.9 nm) with zeta potential ranged from +25.1 ± 1.5 to +39.4 ± 0.5 mV, and high entrapment (>95%) and binding efficiencies. Similarly, CS-TPP nanoparticles showed better siRNA protection during storage at 4°C and as determined by serum protection assay. TEM micrographs revealed the assorted morphology of CS-TPP-siRNA nanoparticles in contrast to irregular morphology displayed by CS-DS-siRNA and CS-PGA-siRNA nanoparticles. All siRNA loaded CS-TPP/DS/PGA nanoparticles showed initial burst release followed by sustained release of siRNA. Moreover, all the formulations showed low and concentration-dependent cytotoxicity with human colorectal cancer cells (DLD-1), in vitro. The cellular uptake studies with CS-TPP-siRNA nanoparticles showed successful delivery of siRNA within cytoplasm of DLD-1 cells. The results demonstrate that ionically cross-linked CS-TPP nanoparticles are biocompatible nonviral gene delivery system and generate a solid ground for further optimization studies, for example with regard to steric stabilization and targeting.

Original languageEnglish
Article numbere0128963
JournalPLoS One
Volume10
Issue number6
DOIs
Publication statusPublished - 11 Jun 2015

Fingerprint

Chitosan
nanoparticles
tripolyphosphates
small interfering RNA
chitosan
Nanoparticles
Small Interfering RNA
Dextran Sulfate
dextran
glutamic acid
Glutamic Acid
sulfates
Zeta potential
Particle Size
particle size
Particle size
Gene Transfer Techniques
triphosphoric acid
Gelation
Cytotoxicity

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Stability, intracellular delivery, and release of siRNA from chitosan nanoparticles using different cross-linkers. / Raja, Maria Abdul Ghafoor; Katas, Haliza; Wen, Thum Jing.

In: PLoS One, Vol. 10, No. 6, e0128963, 11.06.2015.

Research output: Contribution to journalArticle

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