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mRNA delivery using non-viral PCL nanoparticles.

Ilaria E Palamà1, Barbara Cortese, Stefania D'Amone

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Messenger RNA (mRNA) nanoparticles offer a stable, non-viral gene therapy solution. These poly(ε-caprolactone) nanoparticles protect mRNA and release it effectively inside cells, overcoming delivery challenges.

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Area of Science:

  • Biomaterials Science
  • Nanotechnology
  • Gene Therapy

Background:

  • Messenger RNA (mRNA) is a viable alternative to plasmid DNA for non-viral gene therapy.
  • mRNA's instability in vivo hinders its therapeutic application.
  • Efficient intracellular delivery of mRNA remains a significant challenge.

Purpose of the Study:

  • To develop a nanoparticle system for stable and effective intracellular mRNA delivery.
  • To create a poly(ε-caprolactone) (PCL) nanoparticle formulation for mRNA delivery.
  • To investigate the pH-dependent release kinetics and biocompatibility of the developed nanoparticles.

Main Methods:

  • mRNA-protamine complexes were encapsulated within poly(ε-caprolactone) (PCL) nanoparticles (NPs).
  • Nanoparticle size, structure, and mRNA release kinetics at different pH values (7.4 and 5.0) were analyzed.
  • Cytotoxicity of the NPs was evaluated in NIH 3T3 fibroblasts, HeLa cells, and MG63 osteoblasts.

Main Results:

  • The developed PCL nanoparticles exhibited a core-shell structure with a diameter of approximately 247 nm.
  • mRNA release was pH-dependent, with significantly higher release at endosomal pH (5.0) compared to physiological pH (7.4).
  • The nanoparticles demonstrated no cytotoxicity to tested cell lines over 8 days of incubation.

Conclusions:

  • The developed mRNA-protamine-PCL nanoparticles offer a stable and effective system for intracellular mRNA delivery.
  • The pH-responsive release mechanism enhances therapeutic potential by targeting endosomal environments.
  • These biocompatible PCL nanoparticles address key challenges in mRNA-based gene therapy, paving the way for new therapeutic strategies.