Using Gene Therapy to Protect and Restore Cartilage

Evans, Christopher H.; Ghivizzani, Steven C.; Smith, Peter J.; Shuler, Franklin D.; Zhang, Mi; Robbins, P. D.

doi:10.1097/00003086-200010001-00027

Cited by 87 publications

(57 citation statements)

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“…cDNA was reverse transcribed from 1 μg of total cellular RNA using ThermoScript RT-PCR system (Invitrogen). The amplification primer pair for collagen type II was selected as 5′-CTG CTC GCT GCC GCT GTC CTT-3′ and 5′-AAG GGT CCC AGG TTC TCC ATC-3′, aggrecan was 5′-TGA GGA GGG CTG GAA CAA GTA CC-3′ and 5′-GGA GGT GGT AAT TGC AGG GAACA-3 ′, and β-actin was 5′-CAG GTC ATC ACY ATY GGC AAT GAGC-3′ and 5′-CGG ATG TCM ACG TCA CAC TTC ATGA-3′ [7,42]. PCR reactions were performed using aliquots of the cDNA template with PlatinumPCR Supermix kit (Invitrogen).…”

Section: Gene Expression Analysis For Cartilage Matrix Componentsmentioning

confidence: 99%

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Adenoviral-mediated transfer of TGF-β1 but not IGF-1 induces chondrogenic differentiation of human mesenchymal stem cells in pellet cultures

Kawamura¹,

Chu²,

Sobajima³

et al. 2005

Experimental Hematology

102

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Objective. The objective of the present study was to investigate the potential of application of growth factor genes to induce chondrogenic differentiation of human-derived mesenchymal stem cells (MSCs). The growth factor genes evaluated in the present study were transforming growth factor 1 (TGF-β1) and insulin-like growth factor 1 (IGF-1).Methods. Human MSCs were transduced with the adenoviral vectors carrying either TGF-β1 or IGF-1 (AdTGF-β1 and AdIGF-1 respectively) or a combination of both growth factor genes at different multiplicities of infection (MOI) and were then made into pellets. Pellets were also made from nontransduced cells and maintained in culture medium supplemented with 10 ng/mL of TGF-β1. At specified time points, histological analysis, cartilage matrix gene expression, and immunofluorescence were performed to determine the extent of chondrogenic differentiation.Results. MSCs transduced with the AdTGF-β1 demonstrated robust chondrogenic differentiation, while those made from AdIGF-1 did not. AdTGF-β1 pellets demonstrated aggrecan gene expression as early as day 3 of pellet culture, while type II collagen gene expression was detected by day 10 of culture. The AdIGF-1, alone or in combination with TGF-β1 pellets, did not show any type II collagen gene expression at any time point. By immunofluoresecence, type X collagen was distributed throughout the matrix in TGF-β1 protein pellets while the growth factor gene pellets displayed scant staining. Conclusion.The results suggest that sustained administration of TGF-β1 may be more effective in suppressing terminal differentiation than intermittent dosing and thus effective for cartilage repair.

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Section: Gene Expression Analysis For Cartilage Matrix Componentsmentioning

confidence: 99%

“…As an attempt to circumvent the problems associated with the above-mentioned approaches for cartilage repair, novel approaches that include gene and stem cell therapies are being evaluated [6,7]. Adult-derived stem cells that can be harvested from bone, fat, or muscle offer alternative sources for chondrocytes [8][9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Adenoviral-mediated transfer of TGF-β1 but not IGF-1 induces chondrogenic differentiation of human mesenchymal stem cells in pellet cultures

Kawamura¹,

Chu²,

Sobajima³

et al. 2005

Experimental Hematology

102

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show abstract

“…In this respect, there are two obvious targets for intra-articular gene delivery, the synovial lining of the joint, and the articular cartilage (Figure 2). 80 All noncartilagenous, intra-articular surfaces are synovial. The synovial lining has a large surface area, and because it is in direct contact with the joint space, secreted products can diffuse from their synovial origin to other intra-articular tissues, including the cartilage.…”

Section: Strategies For Oa Gene Therapymentioning

confidence: 99%

Osteoarthritis gene therapy

et al. 2004

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“…[7][8][9][10][11] Sustained, controlled administration of the appropriate protein cocktail to articular tissues could be used to improve approaches to the treatment of injured cartilage. 12 This could occur either by fostering differentiation into and/or maintenance of the appropriate chondrocyte phenotype for synthesis of repair tissue that functionally resembles normal articular cartilage, or by protecting the cartilage from the effects of inflammation. Although recombinant proteins are difficult to apply effectively in vivo, gene delivery provides a mechanism to convert specific target cells into factories capable of sustained protein production.…”

Section: Candidate Cdnasmentioning

confidence: 99%