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Related Experiment Videos

Defoliation and plasmid delivery with layer-by-layer coated colloids.

Uta Reibetanz1, Claudia Claus, Elke Typlt

  • 1Institute of Medical Physics and Biophysics, University of Leipzig, Härtelstrasse 16-18, 04107 Leipzig, Germany.

Macromolecular Bioscience
|February 4, 2006
PubMed
Summary

Protamine-coated silica particles efficiently enter cells, releasing plasmids for protein expression. This demonstrates effective gene delivery via non-receptor-mediated endocytosis and particle defoliation.

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Area of Science:

  • Biomaterials Science
  • Cell Biology
  • Gene Delivery

Background:

  • Polyelectrolyte multilayers (PEM) offer versatile surface modification for nanoparticles.
  • Cellular uptake and intracellular fate of coated nanoparticles are crucial for drug and gene delivery applications.

Purpose of the Study:

  • To investigate the cellular uptake, defoliation, and plasmid delivery efficiency of polyelectrolyte multilayer coated silica nanoparticles.
  • To determine the optimal layer configuration for nanoparticle uptake and payload release within cells.

Main Methods:

  • Confocal microscopy and flow cytometry were employed to track nanoparticle uptake and intracellular processes.
  • HEK 293T cells were incubated with silica particles coated with protamine and dextran sulfate in varying layer orders.

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  • Gene expression of enhanced green fluorescence protein (pEGFP-C1) and red fluorescence protein (pDsRed1-N1) was analyzed to confirm plasmid delivery.

    • Main Results:

    • Optimal cellular uptake was achieved when protamine served as the outermost layer of the polyelectrolyte coating.
    • Nanoparticle uptake was observed to proceed via a non-receptor-mediated endocytotic pathway.
    • Successful defoliation of the polyelectrolyte shell within the cells was confirmed by the release of plasmids and subsequent protein expression, along with the observation of fluorescent layer debris.

    Conclusions:

    • Protamine-coated silica nanoparticles demonstrate efficient cellular internalization and payload release.
    • The study validates polyelectrolyte multilayer coated nanoparticles as a viable system for intracellular plasmid delivery.
    • Defoliation of the coating and release of cargo into the cytoplasm are key events for successful gene expression.