Therapeutic Effects of rAAV-Mediated Concomittant Gene Transfer and Overexpression of TGF-β and IGF-I on the Chondrogenesis of Human Bone-Marrow-Derived Mesenchymal Stem Cells

Application of chondroreparative gene vectors in cartilage defects is a powerful approach to directly stimulate the regenerative activities of bone-marrow-derived mesenchymal stem cells (MSCs) that repopulate such lesions. Here, we investigated the ability of combined recombinant adeno-associated vi...

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Bibliographic Details
Main Authors: Stephanie Morscheid, Ana Rey-Rico, Gertrud Schmitt, Henning Madry, Magali Cucchiarini, Jagadeesh Kumar Venkatesan
Format: Article
Language:English
Published: MDPI AG 2019-05-01
Series:International Journal of Molecular Sciences
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Online Access:https://www.mdpi.com/1422-0067/20/10/2591
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Summary:Application of chondroreparative gene vectors in cartilage defects is a powerful approach to directly stimulate the regenerative activities of bone-marrow-derived mesenchymal stem cells (MSCs) that repopulate such lesions. Here, we investigated the ability of combined recombinant adeno-associated virus (rAAV) vector-mediated delivery of the potent transforming growth factor beta (TGF-&#946;) and insulin-like growth factor I (IGF-I) to enhance the processes of chondrogenic differentiation in human MSCs (hMSCs) relative to individual candidate treatments and to reporter (<i>lacZ</i>) gene condition. The rAAV-hTGF-&#946; and rAAV-hIGF-I vectors were simultaneously provided to hMSC aggregate cultures (TGF-&#946;/IGF-I condition) in chondrogenic medium over time (21 days) versus TGF-&#946;/<i>lacZ</i>, IGF-I/<i>lacZ</i>, and <i>lacZ</i> treatments at equivalent vector doses. The cultures were then processed to monitor transgene (co)-overexpression, the levels of biological activities in the cells (cell proliferation, matrix synthesis), and the development of a chondrogenic versus osteogenic/hypertrophic phenotype. Effective, durable co-overexpression of TGF-&#946; with IGF-I via rAAV enhanced the proliferative, anabolic, and chondrogenic activities in hMSCs versus <i>lacZ</i> treatment and reached levels that were higher than those achieved upon single candidate gene transfer, while osteogenic/hypertrophic differentiation was delayed over the period of time evaluated. These findings demonstrate the potential of manipulating multiple therapeutic rAAV vectors as a tool to directly target bone-marrow-derived MSCs in sites of focal cartilage defects and to locally enhance the endogenous processes of cartilage repair.
ISSN:1422-0067