On the Influence of Fabrication Methods and Materials for mRNA-LNP Production: From Size and Morphology to Internal Structure and mRNA Delivery Performance In Vitro and In Vivo.

Lipid nanoparticle (LNP) remains the most advanced platform for messenger RNA (mRNA) delivery. To date, mRNA LNPs synthesis is mostly performed by mixing lipids and mRNA with microfluidics. In this study, we developed a cost-effective microfluidics for synthesizing mRNA LNPs. It allows to fine-tune the LNPs characteristics without compromising LNP properties. We compared it with a commercial device (NanoAssemblr) and ethanol injection and investigated the influence of manufacturing conditions on the performance of mRNA LNPs. LNPs prepared by ethanol injection exhibited broader size distributions and more inhomogeneous internal structure (e.g., bleb-like substructures), while other LNPs showed uniform structures with dense cores. Small angel X-ray scattering (SAXS) data indicate a tighter interaction between mRNA and lipids within LNPs synthesized by custom device, compared to LNPs produced by NanoAssemblr. Moreover, the better transfection efficiency of polysarcosine (pSar)-modified LNPs may be attributed to a higher surface roughness than that of PEGylated LNPs. The manufacturing approach showed modest influence on mRNA expression in vivo. In summary, the home-developed cost-effective microfluidic device can synthesize LNPs and represents a potent alternative to NanoAssemblr. The preparation methods showed notable effect on LNPs' structure but a minor influence on mRNA delivery in vitro and in vivo. This article is protected by copyright. All rights reserved.

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This article is protected by copyright. All rights reserved.