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

Peter Ueblacker, Bettina Wagner, Stephan Vogt, Gian Salzmann, Gabi Wexel, Achim Krüger, Christian Plank, Thomas Brill, Karin Specht, Tilla Hennig, Ulrike Schillinger, Andreas B. Imhoff, Vladimir Martinek, Bernd Gansbacher

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

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 β-galactosidase activity could be determined in the majority of implanted chondrocytes in the osteochondral defects after 4 weeks.

Original languageEnglish
Pages (from-to)4480-4487
Number of pages8
JournalBiomaterials
Volume28
Issue number30
DOIs
StatePublished - Oct 2007

Keywords

  • Chondrocytes
  • Gene transfer
  • Retrovirus
  • Tissue-engineering
  • Transplantation

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