In vivo analysis of retroviral gene transfer to chondrocytes within collagen scaffolds for the treatment of osteochondral defects

Biomaterials. 2007 Oct;28(30):4480-7. doi: 10.1016/j.biomaterials.2007.06.027. Epub 2007 Jul 13.


To examine a retroviral gene transfer to chondrocytes in vitro and in vivo in tissue-engineered cell-collagen constructs articular chondrocytes from rabbits and humans were isolated and transduced with VSV.G pseudotyped murine leukemia virus-derived retroviral vectors. Viral supernatants were generated by transient transfection of 293T cells using the pBullet retroviral vector carrying the nlslacZ gene, a Moloney murine leukemia virus gag/pol plasmid and a VSV.G coding plasmid. Transduction efficiency was analyzed by fluorescence-activated-cell-sorter analysis and transduced autologous chondrocytes from rabbits were seeded on collagen-scaffolds and implanted into osteochondral defects in the patellar groove of the rabbit's femur (n=10). LacZ-expression was analyzed by X-gal staining on total knee explants and histological sections. Retroviral transduction efficiency exceeded 92.3% (SEM+/-3.5%) in rabbit articular chondrocytes, 74.7% (SEM+/-1.8%) in human articular chondrocytes and 52.7% (SEM+/-5.8%) in osteoarthritic human chondrocytes. Reporter gene expression remained high after 15 weeks in 75.7% (SEM+/-8.2%) of transduced rabbit articular chondrocytes. In vivo, intraarticular beta-galactosidase activity could be determined in the majority of implanted chondrocytes in the osteochondral defects after 4 weeks.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cartilage, Articular / cytology
  • Cell Culture Techniques
  • Cell Line
  • Cell Survival
  • Cells, Cultured
  • Chondrocytes / cytology
  • Chondrocytes / metabolism*
  • Chondrocytes / pathology
  • Collagen Type I / metabolism*
  • Female
  • Femur
  • Gene Expression
  • Gene Transfer Techniques*
  • Genetic Therapy
  • Genetic Vectors*
  • Humans
  • Immunohistochemistry
  • Implants, Experimental
  • Kidney / cytology
  • Lac Operon
  • Osteoarthritis / metabolism*
  • Osteoarthritis / pathology
  • Osteoarthritis / therapy
  • Rabbits
  • Retroviridae / genetics
  • Time Factors
  • Tissue Engineering / methods
  • Transduction, Genetic
  • Transplantation, Autologous
  • beta-Galactosidase / genetics


  • Collagen Type I
  • beta-Galactosidase