rAAV-mediated overexpression of TGF-β via vector delivery in polymeric micelles stimulates the biological and reparative activities of human articular chondrocytes in vitro and in a human osteochondral defect model

Abstract

Recombinant adeno-associated virus (rAAV) vectors are clinically adapted vectors to durably treat human osteoarthritis (OA). Controlled delivery of rAAV vectors via polymeric micelles was reported to enhance the temporal and spatial presentation of the vectors into their targets. Here, we tested the feasibility of delivering rAAV vectors via poly (ethylene oxide) (PEO) and poly (propylene oxide) (PPO) (poloxamer and poloxamine) polymeric micelles as a means to overexpress the therapeutic factor transforming growth factor-beta (TGF-β) in human OA chondrocytes and in experimental human osteochondral defects. Application of rAAV-human transforming growth factor-beta using such micelles increased the levels of TGF-β transgene expression compared with free vector treatment. Overexpression of TGF-β with these systems resulted in higher proteoglycan deposition and increased cell numbers in OA chondrocytes. In osteochondral defect cultures, a higher deposition of type-II collagen and reduced hypertrophic events were noted. Delivery of therapeutic rAAV vectors via PEO-PPO-PEO micelles may provide potential tools to remodel human OA cartilage.

Biomaterial-Guided Recombinant Adeno-associated Virus Delivery from Poly(Sodium Styrene Sulfonate)-Grafted Poly(ɛ-Caprolactone) Films to Target Human Bone Marrow Aspirates

“…[17][18][19] While work thus far emphasized on delivering shortlived nonviral [20][21][22][23][24][25][26][27][28][29][30] and potentially oncogenic lentiviral vectors, [31][32][33][34] there is still little information on transferring the more effective, clinically preferred recombinant adenoassociated virus (rAAV) vectors using biomaterials. 17,19,[35][36][37][38] Interestingly, most studies focused on the value of hydrogel systems for rAAV-based cartilage 6 regenerative medicine (fibrin, alginate, poloxamers, poloxamines, self-assembling peptides, polypseudorotaxanes), [39][40][41][42][43][44][45][46] while no evidence described the potential of mechanically stable solid scaffolds to guide rAAV application in sites of cartilage injury.…”

Enhanced Chondrogenic Differentiation Activities in Human Bone Marrow Aspirates via sox9 Overexpression Mediated by pNaSS-Grafted PCL Film-Guided rAAV Gene Transfer

“…Scaffold-assisted gene therapy is a powerful tool to durably and safely enhance the processes of cartilage repair [10][11][12] via single-step, controlled delivery of clinically adapted rAAV gene transfer vectors [25][26][27][28][29][30][31][32][33]. In the current work, we tested the ability of mechanically stable, biocompatible PCL scaffolds functionalized with a bioactive pNaSS molecule to transfer rAAV vectors coding for the cartilage-specific SOX9 transcription factor to human bone marrow aspirates as a means to generate more efficient, non-invasive systems to treat focal cartilage lesions compared with a less stable scaffold-free vector application lacking scaffolding benefits for the cell targets [42] or with complex, indirect transplantation of rAAV-modified aspirates via such materials [43].…”

“…As a matter of fact, a number of studies reported the potential of applying rAAV for experimental cartilage research via hydrogel systems (alginate, fibrin, poloxamers/poloxamines, self-assembling peptides, polypseudorotaxanes) [25][26][27][28][29][30][31][32][33] while there is still little information on the potential benefits of solid, mechanically more stable biomaterials that may provide scaffolding and stability to the target cells [34] for rAAV-mediated gene transfer. In this regard, we recently provided evidence that biocompatible solid polyester poly(ε-caprolactone) (PCL) [35], an aliphatic polyester approved by the FDA [36,37], further grafted with poly(sodium styrene sulfonate) (pNaSS) to activate reparative cellular responses [38] is capable of supporting the delivery of reporter rAAV gene vectors to effectively modify human bone marrow aspirates [39].…”

Cyclodextrin Cationic Polymer-Based Nanoassemblies to Manage Inflammation by Intra-Articular Delivery Strategies

“…hMSCs were seeded in 24-well plates (2 × 10 4 cells/well) with growth medium for 12 h at 37 °C under 5% CO 2 . PolyCD-based nanoassemblies were then directly added to the cultures and live fluorescence was monitored in the samples by fluorescent microscopy using a rhodamine filter set (568 nm; Olympus CKX41; Hamburg, Germany) [ 55 , 57 ].…”