Chip-Free Microscale-Incubator-Based Synthesis of Chitosan-Based Gene Silencing Nanoparticles

Noemi Gaglianone, Michael Lykke Hvam, Hüsnü Aslan, Mingdong Dong, Ken Howard, Yi-Ping Ho

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review


The development of polymer-based nanoparticles to ferry siRNA continues to evolve. It is becoming increasingly apparent that gene silencing nanoparticles produced by conventional bulk manufacturing techniques often exhibit physicochemical heterogeneity within and between batches that can affect the biological performance. Here a new facile and robust "chip-free" method is presented, termed chip-free agitation-generated droplets (CAD) preparation, using chitosan-based gene silencing nanoparticles as an example. The CAD-prepared silencing particles, in comparison to the particles prepared by the conventional bulk protocol, exhibit lower surface charge (9 mV vs 21 mV at N/P = 5), higher stability (≈40% higher binding affinity and up to 30% less morphological deformation), and are less prone to aggregation measured by nanoparticle tracking analysis over a period of one month. Furthermore, these physical attributes contribute up to 19% higher in cell viability at N/P = 5, while the gene silencing of enhanced green fluorescent protein remains constant in a human cell line. Control of particle properties is necessary to advance siRNA-based delivery; the CAD preparation represents a physical complement to chemical design modifications, which can be readily transferred among research labs and utilized for alternative polymer systems. A new, facile, and robust "chip-free" method for nanoparticle preparation is presented using chitosan-based gene-silencing nanoparticles as an example. The prepared particles exhibit low surface charge, high stability and are less prone to aggregation. These physical attributes are observed to increase cell viability, whilst the gene silencing remains unaltered in a human cell line.

Original languageEnglish
JournalParticle & Particle Systems Characterization
Pages (from-to)279-285
Number of pages6
Publication statusPublished - May 2016


  • microscale counterion interactions
  • nanoparticles
  • siRNA


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