Charge‐Reversed Exosomes for Targeted Gene Delivery to Cartilage for Osteoarthritis Treatment

Abstract Gene therapy has the potential to facilitate targeted expression of therapeutic proteins to promote cartilage regeneration in osteoarthritis (OA). The dense, avascular, aggrecan‐glycosaminoglycan (GAG) rich negatively charged cartilage, however, hinders their transport to reach chondrocytes in effective doses. While viral vector mediated gene delivery has shown promise, concerns over immunogenicity and tumorigenic side‐effects persist. To address these issues, this study develops surface‐modified cartilage‐targeting exosomes as non‐viral carriers for gene therapy. Charge‐reversed cationic exosomes are engineered for mRNA delivery by anchoring cartilage targeting optimally charged arginine‐rich cationic motifs into the anionic exosome bilayer by using buffer pH as a charge‐reversal switch. Cationic exosomes penetrated through the full‐thickness of early‐stage arthritic human cartilage owing to weak‐reversible ionic binding with GAGs and efficiently delivered the encapsulated eGFP mRNA to chondrocytes residing in tissue deep layers, while unmodified anionic exosomes do not. When intra‐articularly injected into destabilized medial meniscus mice knees with early‐stage OA, mRNA loaded charge‐reversed exosomes overcame joint clearance and rapidly penetrated into cartilage, creating an intra‐tissue depot and efficiently expressing eGFP; native exosomes remained unsuccessful. Cationic exosomes thus hold strong translational potential as a platform technology for cartilage‐targeted non‐viral delivery of any relevant mRNA targets for OA treatment.