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Self-Assembled Polypeptide Nanogels with Enzymatically Transformable Surface as a Small Interfering RNA Delivery

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Enzyme-responsive nanogels offer a novel platform for delivering therapeutic macromolecules like small interfering RNA (siRNA). These nanogels shield charges for stability, then release them upon enzyme exposure for efficient cellular delivery and gene silencing.

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Area of Science:

  • Biomaterials Science
  • Nanotechnology
  • Drug Delivery Systems

Background:

  • Cationic nanogels are promising for macromolecule delivery but face instability due to electrostatic interactions.
  • Existing charge-shielding methods like PEGylation reduce delivery efficiency.
  • Biodegradable polymer coatings are synthetically challenging to control.

Purpose of the Study:

  • To develop and evaluate enzyme-responsive nanogels for enhanced therapeutic cargo delivery.
  • To create a nanogel system that shields cationic charges for stability and exposes them upon enzymatic degradation for cellular uptake.
  • To assess the potential of these nanogels as carriers for small interfering RNA (siRNA).

Main Methods:

  • Synthesized maltopentaose functionalized cholesteryl poly(l-lysine) nanogels (CbAmyPL).
  • Utilized tandem enzymatic polymerization with glycogen phosphorylase and glycogen branching enzyme to create a branched amylose shell.
  • Characterized nanogel properties (ζ potential) and siRNA complex stability using Foster resonance energy transfer (FRET).
  • Evaluated nanogel cytotoxicity and siRNA delivery efficiency in murine renal carcinoma (Renca) cells with and without α-amylase.

Main Results:

  • CbAmyPL nanogels exhibited neutral ζ potential, which became positive after α-amylase degradation.
  • Nanogels formed stable complexes with siRNA, resistant to serum proteins and enzymatic degradation.
  • CbAmyPL/siRNA complexes demonstrated enhanced vascular endothelial growth factor (VEGF) mRNA knockdown (50%) in Renca cells in the presence of α-amylase, compared to without enzyme (20%).
  • The nanogels showed no significant cytotoxicity.

Conclusions:

  • Enzyme-responsive nanogels provide a stable and effective platform for siRNA delivery.
  • The proposed enzymatic functionalization strategy overcomes limitations of conventional nanocarrier coatings.
  • This technology holds promise for advancing therapeutic macromolecule delivery applications.