Ex vivo gene transfer to chondrocytes in full-thickness articular cartilage defects: a feasibility study

Kang, Richard W.; Marui, Takashi; Ghivizzani, Steven C.; Nita, Ioana M.; Georgescu, Helga I.; Suh, Jun-Kyo Francis; Robbins, Paul D.; Evans, Christopher H.

doi:10.1016/s1063-4584(97)80007-6

Cited by 126 publications

(74 citation statements)

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“…Although feasible, our initial experiments demonstrated several shortfalls of this approach, particularly with regard to consistency of application and vector containment. While this direct method greatly streamlined the process of gene delivery to osteochondral defects relative to ex vivo strategies, 23 control over the process was sacrificed. We routinely observed 100-fold variations in the level of transgenic expression between implants.…”

Section: Discussionmentioning

confidence: 99%

“…28 In a previous study, we found that a biologically compatible matrix implanted into an osteochondral defect in the femoral condyle of the rabbit will become increasingly cellular with time. 23 Since chondrocytes from the surrounding tissue are unlikely to traffic into the lesion, these observations provided evidence that cells from the bone marrow could infiltrate and colonize an implanted matrix. They likewise suggested that it may be possible to deliver transgenes directly to the cells that infiltrate osteochondral lesions from the underlying marrow.…”

Section: Introductionmentioning

confidence: 95%

“…22 Preclinical studies of gene transfer for cartilage repair have typically favored ex vivo approaches because they deliver chondrogenic cells and their expressed gene products to the defect site. In this regard, chondrocytes, periosteal cells and MSCs have been tested as vehicles for gene delivery in animal models, [23][24][25][26][27] and while there has been evidence of cartilage repair, clinical application would require harvesting and culturing of autologous cells for each patient. From our clinical experience with ex vivo gene delivery to joints, it has become evident that the facilities, labor, time and, therefore, expense of these procedures may make this approach impractical for mainstream application.…”

Section: Introductionmentioning

confidence: 99%

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Gene delivery to cartilage defects using coagulated bone marrow aspirate

et al. 2004

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The long-term goal of the present study is to develop a clinically applicable approach to enhance natural repair mechanisms within cartilage lesions by targeting bone marrow-derived cells for genetic modification. To determine if bone marrow-derived cells infiltrating osteochondral defects could be transduced in situ, we implanted collagen-glycosaminoglycan (CG) matrices preloaded with adenoviral vectors containing various marker genes into lesions surgically generated in rabbit femoral condyles. Analysis of the recovered implants showed transgenic expression up to 21 days; however, a considerable portion was found in the synovial lining, indicating leakage of the vector and/or transduced cells from the matrix. As an alternative medium for gene delivery, we investigated the feasibility of using coagulated bone marrow aspirates. Mixture of an adenoviral suspension with the fluid phase of freshly aspirated bone marrow resulted in uniform dispersion of the vector throughout, and levels of transgenic expression in direct proportion to the density of nucleated cells in the ensuing clot. Furthermore, cultures of mesenchymal progenitor cells, previously transduced ex vivo with recombinant adenovirus, were readily incorporated into the coagulate when mixed with fresh aspirate. These vector-seeded and cell-seeded bone marrow clots were found to maintain their structural integrity following extensive culture and maintained transgenic expression in this manner for several weeks. When used in place of the CG matrix as a gene delivery vehicle in vivo, genetically modified bone marrow clots were able to generate similarly high levels of transgenic expression in osteochondral defects with better containment of the vector within the defect. Our results suggest that coagulates formed from aspirated bone marrow may be useful as a means of gene delivery to cartilage and perhaps other musculoskeletal tissues. Cells within the fluid can be readily modified with an adenoviral vector, and the matrix formed from the clot is completely natural, native to the host and is the fundamental platform on which healing and repair of mesenchymal tissues is based.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

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Gene delivery to cartilage defects using coagulated bone marrow aspirate

et al. 2004

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“…Chondrocytes cultured in monolayer are receptive to transduction using the more common viral vectors, including adenovirus, [47][48][49] oncoretrovirus (MoMLV), 50 VSV-G pseudotyped lentivirus 33 and AAV. 44,48,51 Although chondrocytes have been somewhat resistant to transfection with plasmid DNA, formulation with certain commercially available lipid-based reagents such as FuGENE6t 52,53 and Lipofectint has been found to enhance the efficiency of DNA uptake, as has pretreatment of the cells with hyaluronidase 54 and mild detergent.…”

Section: Chondrocytes As Targets For Gene Deliverymentioning

confidence: 99%

“…Initial in vivo studies by Kang et al 50 showed that allogenic chondrocytes transduced with a retroviral vector encoding a marker gene, when embedded in a collagen gel and delivered to osteochondral defects in rabbits, could support transgenic expression for several weeks. Similar results have been obtained with chondrocyte-seeded collagen type I matrices following genetic modification of the cells with adenovirus.…”

Section: Chondrocytes As Targets For Gene Deliverymentioning

confidence: 99%