The genetic modification of freshly aspirated bone marrow may provide convenient tools to enhance the regenerative capacities of\ncartilage defects compared with the complex manipulation of isolated progenitor cells. In the present study, we examined the\nability and safety of recombinant adeno-associated virus (rAAV) serotype 2 vectors to deliver various reporter gene sequences in\nprimary human bone marrow aspirates over time without altering the chondrogenic processes in the samples. The results\ndemonstrate that successful rAAV-mediated gene transfer and expression of the lacZ and red fluorescent protein marker genes\nwere achieved in transduced aspirates at very high efficiencies (90ââ?¬â??94%) and over extended periods of time (up to 125 days) upon\ntreatment with hirudin, an alternative anticoagulant that does not prevent the adsorption of the rAAV-2 particles at the surface of\ntheir targets compared with heparin. Application of rAAV was safe, displaying neither cytotoxic nor detrimental effects on the\ncellular and proliferative activities or on the chondrogenic processes in the aspirates especially using an optimal dose of\n0.5 mg ml? 1 hirudin, and application of the potent SOX9 transcription factor even enhanced these processes while counteracting\nhypertrophic differentiation. The current findings demonstrate the clinical value of this class of vector to durably and safely modify\nbone marrow aspirates as a means to further develop convenient therapeutic approaches to improve the healing of cartilage\ndefects.
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