WNT3A‐loaded exosomes enable cartilage repair (original) (raw)

2021, Journal of Extracellular Vesicles

Cartilage defects repair poorly. Recent genetic studies suggest that WNT3a may contribute to cartilage regeneration, however the dense, avascular cartilage extracellular matrix limits its penetration and signalling to chondrocytes. Extracellular vesicles actively penetrate intact cartilage. This study investigates the effect of delivering WNT3a into large cartilage defects in vivo using exosomes as a delivery vehicle. Exosomes were purified by ultracentrifugation from conditioned medium of either L-cells overexpressing WNT3a or control un-transduced L-cells, and characterized by electron microscopy, nanoparticle tracking analysis and marker profiling. WNT3a loaded on exosomes was quantified by western blotting and functionally characterized in vitro using the SUPER8TOPFlash reporter assay and other established readouts including proliferation and proteoglycan content. In vivo pathway activation was assessed using TCF/Lef:H2B-GFP reporter mice. Wnt3a loaded exosomes were injected into the knees of mice, in which large osteochondral defects were surgically generated. The degree of repair was histologically scored after 8 weeks. WNT3a was successfully loaded on exosomes and resulted in activation of WNT signalling in vitro. In vivo, recombinant WNT3a failed to activate WNT signalling in cartilage, whereas a single administration of WNT3a loaded exosomes activated canonical WNT signalling for at least one week, and eight weeks later, improved the repair of osteochondral defects. WNT3a assembled on exosomes, is efficiently delivered into cartilage and contributes to the healing of osteochondral defects. K E Y WO R D S cartilage, drug delivery, exosomes, joint repair, WNT3a  INTRODUCTION Cartilage defects are present in 61% of patients undergoing knee arthroscopy (Curl et al., 1997; Hjelle et al., 2002), can be disabling, and have the potential to progress into osteoarthritis (Dell'accio & Vincent, 2010). Cell based approaches to cartilage repair are effective, but, due to the largely autologous nature of the transplanted cells, such procedures are costly and laborious. A number of novel biological agents are being developed which harness and support the endogenous capacity of joint to repair (Sherwood, 2019). One major limitation in delivering such agents (often macromolecules) is the avascular nature of the cartilage tissue and the dense and strongly negatively charged extracellular matrix. Antibodies only penetrate the very superficial layers This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.