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Local release of siRNA using polyplex-loaded thermosensitive hydrogels.

Lies A L Fliervoet1, Heyang Zhang, Emma van Groesen

  • 1Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences (UIPS), Faculty of Science, Utrecht University, PO Box 80082, 3508 TB Utrecht, the Netherlands. T.Vermonden@uu.nl.

Nanoscale
|May 6, 2020
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Summary

This study developed a thermosensitive hydrogel for sustained local delivery of small interfering RNA (siRNA) polyplexes. The hydrogel system demonstrated controlled release and maintained siRNA

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Area of Science:

  • Biomaterials Science
  • Polymer Chemistry
  • Gene Therapy

Background:

  • Clinical translation of RNA interference (RNAi) therapies is limited by challenges in delivering therapeutic nucleic acids like small interfering RNA (siRNA) locally without off-target effects.
  • Hydrogels offer a promising solution for localized and sustained release of siRNA cargo.
  • Developing effective delivery systems is crucial for advancing RNAi-based therapeutics.

Purpose of the Study:

  • To investigate the formation of siRNA polyplexes using poly(2-dimethylaminoethyl methacrylate) (PDMAEMA)-based polymers.
  • To load these polyplexes into a thermosensitive hydrogel for controlled local siRNA release.
  • To evaluate the stability, release kinetics, and biological activity of siRNA delivered via the hydrogel system.

Main Methods:

  • Formation and characterization of siRNA polyplexes with multifunctional NPD and control PD polymers.
  • Loading of polyplexes into a thermosensitive PNIPAM-PEG-PNIPAM hydrogel.
  • In vitro release studies of polyplexes and free siRNA from the hydrogel.
  • Assessment of polyplex integrity post-release.
  • Evaluation of the biological activity of released polyplexes through siRNA-induced gene silencing in FaDu cells.

Main Results:

  • Small polyplexes (10-20 nm) were formed at N/P charge ratios of 5 or higher.
  • Hydrogel formulation resulted in significantly more controlled and sustained siRNA release (128 hours) compared to free siRNA (50 hours).
  • Released polyplexes maintained their size and biological activity, effectively transfecting cells and inducing luciferase silencing.

Conclusions:

  • An injectable, thermosensitive hydrogel system was successfully developed for local and sustained siRNA delivery.
  • This hydrogel formulation enhances siRNA stability and controls its release profile.
  • The system shows potential for various therapeutic applications, including tumor treatment, by improving siRNA delivery efficacy.