Enhanced Chondrogenic Differentiation Activities in Human Bone Marrow Aspirates via Overexpression Mediated by pNaSS-Grafted PCL Film-Guided rAAV Gene Transfer.

BACKGROUND

The delivery of therapeutic genes in sites of articular cartilage lesions using non-invasive, scaffold-guided gene therapy procedures is a promising approach to stimulate cartilage repair while protecting the cargos from detrimental immune responses, particularly when targeting chondroreparative bone marrow-derived mesenchymal stromal cells in a natural microenvironment like marrow aspirates.

METHODS

Here, we evaluated the benefits of providing a sequence for the cartilage-specific sex-determining region Y-type high-mobility group box 9 (SOX9) transcription factor to human marrow aspirates via recombinant adeno-associated virus (rAAV) vectors delivered by poly(ε-caprolactone) (PCL) films functionalized via grafting with poly(sodium styrene sulfonate) (pNaSS) to enhance the marrow chondrogenic potential over time.

RESULTS

Effective overexpression was observed in aspirates treated with pNaSS-grafted or ungrafted PCL films coated with the candidate rAAV-FLAG-h (FLAG-tagged rAAV vector carrying a human gene sequence) vector for at least 21 days relative to other conditions (pNaSS-grafted and ungrafted PCL films without vector coating). Overexpression of via rAAV /pNaSS-grafted or ungrafted PCL films led to increased biological and chondrogenic differentiation activities (matrix deposition) in the aspirates while containing premature osteogenesis and hypertrophy without impacting cell proliferation, with more potent effects noted when using pNaSS-grafted films.

CONCLUSIONS

These findings show the benefits of targeting patients' bone marrow via PCL film-guided therapeutic rAAV () delivery as an off-the-shelf system for future strategies to enhance cartilage repair in translational applications.