Sustained transgene expression in cartilage defects in vivo after transplantation of articular chondrocytes modified by lipid-mediated gene transfer in a gel suspension delivery system

Henning Madry, Magali Cucchiarini, Ute Stein, Klaus Remberger, Michael D. Menger, Dieter Kohn, Stephen Trippel

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Background: Genetically modified chondrocytes may be able to modulate articular cartilage repair. To date, transplantation of modified chondrocytes into cartilage defects has been restricted to vital vectors. We tested the hypothesis that a recombinant gene can be delivered to sites of cartilage damage in vivo using chondrocytes transfected by a lipid-mediated gene transfer method. Methods: Isolated lapine articular chondrocytes were transfected with an expression plasmid vector carrying the P. pyralis luciferase gene using the reagent FuGENE 6. Transfected chondrocytes were encapsulated in alginate spheres and implanted into osteochondral defects in the knee joints of rabbits. Results: In vitro, luciferase activity in pCMVLuc-transfected spheres showed an early peak at day 2 post-transfection and remained elevated at day 32, the longest time point evaluated. The number of viable chondrocytes in non-transfected and transfected spheres increased over the period of cultivation. In vivo, luciferase activity was maximal at day 5 post-transfection, declined by day 16, but was still present at day 32. On histological analysis, the alginate-chondrocyre spheres filled the cartilage defects and were surrounded by a fibrous repair tissue composed of spindle-shaped cells. Conclusions: These data demonstrate the successful introduction of articular chondrocytes modified by lipid-mediated gene transfer in a gel suspension delivery system into osteochondral defects and the sustained expression of the transgene in vivo. This method may be used to define the effects of genes involved in cartilage repair and may provide alternative treatments for articular cartilage defects.

Original languageEnglish
Pages (from-to)502-509
Number of pages8
JournalJournal of Gene Medicine
Volume5
Issue number6
DOIs
StatePublished - Jun 2003

Fingerprint

Chondrocytes
Transgenes
Cartilage
Suspensions
Joints
Transplantation
Gels
Lipids
Genes
Luciferases
Articular Cartilage
Transfection
Knee Joint
Plasmids
Rabbits

Keywords

  • Alginate
  • Cartilage defects
  • Chondrocytes
  • Transfection
  • Transplantation

ASJC Scopus subject areas

  • Genetics

Cite this

Sustained transgene expression in cartilage defects in vivo after transplantation of articular chondrocytes modified by lipid-mediated gene transfer in a gel suspension delivery system. / Madry, Henning; Cucchiarini, Magali; Stein, Ute; Remberger, Klaus; Menger, Michael D.; Kohn, Dieter; Trippel, Stephen.

In: Journal of Gene Medicine, Vol. 5, No. 6, 06.2003, p. 502-509.

Research output: Contribution to journalArticle

Madry, Henning ; Cucchiarini, Magali ; Stein, Ute ; Remberger, Klaus ; Menger, Michael D. ; Kohn, Dieter ; Trippel, Stephen. / Sustained transgene expression in cartilage defects in vivo after transplantation of articular chondrocytes modified by lipid-mediated gene transfer in a gel suspension delivery system. In: Journal of Gene Medicine. 2003 ; Vol. 5, No. 6. pp. 502-509.
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abstract = "Background: Genetically modified chondrocytes may be able to modulate articular cartilage repair. To date, transplantation of modified chondrocytes into cartilage defects has been restricted to vital vectors. We tested the hypothesis that a recombinant gene can be delivered to sites of cartilage damage in vivo using chondrocytes transfected by a lipid-mediated gene transfer method. Methods: Isolated lapine articular chondrocytes were transfected with an expression plasmid vector carrying the P. pyralis luciferase gene using the reagent FuGENE 6. Transfected chondrocytes were encapsulated in alginate spheres and implanted into osteochondral defects in the knee joints of rabbits. Results: In vitro, luciferase activity in pCMVLuc-transfected spheres showed an early peak at day 2 post-transfection and remained elevated at day 32, the longest time point evaluated. The number of viable chondrocytes in non-transfected and transfected spheres increased over the period of cultivation. In vivo, luciferase activity was maximal at day 5 post-transfection, declined by day 16, but was still present at day 32. On histological analysis, the alginate-chondrocyre spheres filled the cartilage defects and were surrounded by a fibrous repair tissue composed of spindle-shaped cells. Conclusions: These data demonstrate the successful introduction of articular chondrocytes modified by lipid-mediated gene transfer in a gel suspension delivery system into osteochondral defects and the sustained expression of the transgene in vivo. This method may be used to define the effects of genes involved in cartilage repair and may provide alternative treatments for articular cartilage defects.",
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AB - Background: Genetically modified chondrocytes may be able to modulate articular cartilage repair. To date, transplantation of modified chondrocytes into cartilage defects has been restricted to vital vectors. We tested the hypothesis that a recombinant gene can be delivered to sites of cartilage damage in vivo using chondrocytes transfected by a lipid-mediated gene transfer method. Methods: Isolated lapine articular chondrocytes were transfected with an expression plasmid vector carrying the P. pyralis luciferase gene using the reagent FuGENE 6. Transfected chondrocytes were encapsulated in alginate spheres and implanted into osteochondral defects in the knee joints of rabbits. Results: In vitro, luciferase activity in pCMVLuc-transfected spheres showed an early peak at day 2 post-transfection and remained elevated at day 32, the longest time point evaluated. The number of viable chondrocytes in non-transfected and transfected spheres increased over the period of cultivation. In vivo, luciferase activity was maximal at day 5 post-transfection, declined by day 16, but was still present at day 32. On histological analysis, the alginate-chondrocyre spheres filled the cartilage defects and were surrounded by a fibrous repair tissue composed of spindle-shaped cells. Conclusions: These data demonstrate the successful introduction of articular chondrocytes modified by lipid-mediated gene transfer in a gel suspension delivery system into osteochondral defects and the sustained expression of the transgene in vivo. This method may be used to define the effects of genes involved in cartilage repair and may provide alternative treatments for articular cartilage defects.

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