title: Chondrogenic Differentiation Processes in Human Bone Marrow Aspirates upon rAAV-Mediated Gene Transfer and Overexpression of the Insulin-Like Growth Factor I.
abstract: Direct therapeutic gene transfer in marrow concentrates is an attractive strategy to conveniently enhance the chondrogenic differentiation processes as a means to improve the healing response of damaged articular cartilage upon reimplantation in sites of injury. In the present study, we evaluated the ability of the clinically adapted recombinant adeno-associated virus (rAAV) vectors to mediate overexpression of the insulin-like growth factor I (IGF-I) in human bone marrow aspirates that may modulate the proliferative, anabolic activities, and chondrogenic differentiation potential in such samples in vitro. The results demonstrate that successful, significant rAAV-mediated IGF-I gene transfer and expression were achieved in transduced aspirates (up to 105.9±35.1 pg rhIGF-I/mg total proteins) over time (21 days) at very high levels (∼80% of cells expressing the candidate IGF-I transgene), leading to increased levels of proliferation, matrix synthesis, and chondrogenic differentiation over time compared with the control (lacZ) condition. Treatment with the candidate IGF-I vector also stimulated the hypertrophic and osteogenic differentiation processes in the aspirates, suggesting that the regulation of IGF-I expression through rAAV will be a prerequisite for future translation of the approach in vivo. However, these findings show the possible benefits of this vector class to directly modify marrow concentrates as a convenient tool for strategies that aim at improving the repair of articular cartilage lesions.
authors.affiliation: 1 Center of Experimental Orthopaedics, Saarland University Medical Center , Homburg/Saar, Germany .
authors.forename: Janina
authors.initials: J
authors.lastname: Frisch
authors.affiliation: 1 Center of Experimental Orthopaedics, Saarland University Medical Center , Homburg/Saar, Germany .
authors.forename: Ana
authors.initials: A
authors.lastname: Rey-Rico
authors.affiliation: 1 Center of Experimental Orthopaedics, Saarland University Medical Center , Homburg/Saar, Germany .
authors.forename: Jagadeesh Kumar
authors.initials: JK
authors.lastname: Venkatesan
authors.affiliation: 1 Center of Experimental Orthopaedics, Saarland University Medical Center , Homburg/Saar, Germany .
authors.forename: Gertrud
authors.initials: G
authors.lastname: Schmitt
authors.affiliation: 1 Center of Experimental Orthopaedics, Saarland University Medical Center , Homburg/Saar, Germany .|2 Department of Orthopedic Surgery, Saarland University Medical Center , Homburg/Saar, Germany .
authors.forename: Henning
authors.initials: H
authors.lastname: Madry
authors.affiliation: 1 Center of Experimental Orthopaedics, Saarland University Medical Center , Homburg/Saar, Germany .
authors.forename: Magali
authors.initials: M
authors.lastname: Cucchiarini
mesh_terms: D050156:Adipogenesis; D000368:Aged; D001853:Bone Marrow / Q000378:metabolism*; D002454:Cell Differentiation; D049109:Cell Proliferation; D020219:Chondrogenesis; D000229:Dependovirus / Q000378:metabolism*; D018014:Gene Transfer Techniques; D006801:Humans; D006984:Hypertrophy; D007334:Insulin-Like Growth Factor I / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D013396:Suction; D014161:Transduction, Genetic
chemical_list: D007334:Insulin-Like Growth Factor I