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Formulating and Characterizing Lipid Nanoparticles for Gene Delivery using a Microfluidic Mixing Platform
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RNA lipid nanoparticles stabilized during nebulization through excipient selection.

Kai V Slaughter1,2, Daniela Isaacs-Bernal2,3, Eric N Donders1,2,3

  • 1Institute of Biomedical Engineering, University of Toronto 164 College Street Toronto Ontario M5S 3G9 Canada molly.shoichet@utoronto.ca.

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|June 16, 2025
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Summary
This summary is machine-generated.

This study stabilizes lipid nanoparticles (LNPs) for pulmonary RNA delivery by optimizing nebulization buffer composition. Adjusting pH, adding poloxamer 188, and including glucose enhances LNP stability and maintains RNA bioactivity for effective inhalation therapies.

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Area of Science:

  • Biotechnology
  • Nanomedicine
  • Pulmonary Drug Delivery

Background:

  • Lipid nanoparticles (LNPs) are promising for pulmonary RNA delivery.
  • Nebulization subjects LNPs to shear forces, causing destabilization and cargo loss.

Purpose of the Study:

  • To develop a generalizable method for stabilizing LNPs during nebulization.
  • To optimize nebulization buffer composition for improved LNP stability and RNA delivery.

Main Methods:

  • Investigated the impact of buffer composition (pH, additives like poloxamer 188, and osmolarity agents like glucose) on LNP characteristics.
  • Assessed nanoparticle size, RNA encapsulation efficiency, and LNP recovery.
  • Evaluated the bioactivity of nebulized LNPs through cellular uptake and siRNA delivery assays.

Main Results:

  • A pH 5.0 citrate buffer minimized encapsulated RNA loss.
  • Poloxamer 188 addition maintained LNP size and improved material recovery.
  • Glucose ensured an iso-osmotic solution, crucial for LNP stability.
  • Nebulized LNPs retained bioactivity, enabling functional siRNA delivery to target cells.

Conclusions:

  • Optimizing nebulization buffer composition is a versatile strategy to stabilize inhalable LNP-based RNA therapies.
  • This approach enhances LNP integrity and preserves therapeutic RNA cargo during aerosolization.
  • The findings support the development of effective inhaled RNA-based treatments for pulmonary disorders.