Hybrid lipid nanoparticles derived from human mesenchymal stem cell extracellular vesicles by microfluidic-sonication for collagen I mRNA delivery to human tendon progenitor stem cells

Tendon degeneration remains an intricate pathological process characterized by the coexistence of multiple dysregulated homeostasis processes, including the increase in collagen III production in comparison with collagen I. Mesenchymal stem cells-derived extracellular vesicles (MSC-EVs) remain a promising therapeutic tool thanks to their pro-regenerative properties and applicability as drug delivery systems, despite their drug loading limitations. Herein, we developed MSC-EVs derived hybrid lipid nanoparticles (MSC-Hyb NPs) using a microfluidic-sonication technique as an alternative platform for the delivery of collagen type I (COL 1A1) mRNA in pathological TSPCs. The produced MSC-Hyb NPs owned LNPs-like physicochemical characteristics with 178.6 nm size and 0.245 PDI values. Moreover, MSC-Hyb NPs encapsulated mRNA and included EVs-derived surface proteins such as CD63, CD81 and CD144. MSC-Hyb NPs remained highly biocompatible with TSPCs and proved to be functional mRNA delivery agents with certain limitations in comparison with lipid nanoparticles (LNPs). In vitro efficacy studies in TSPCs showed a 2-fold increase in procollagen type I carboxy-terminal peptide production comparable with the effect caused by LNPs. Therefore, our work provides an alternative production method for MSC-EVs derived hybrid NPs and supports their potential use as drug delivery systems for tendon regeneration.