Regeneration of hyaline articular cartilage with irradiated transforming growth factor beta1-producing fibroblasts.

The regeneration of hyaline articular cartilage by cell-mediated gene therapy using transforming growth factor beta(1) (TGF-beta(1))-producing fibroblasts (NIH 3T3-TGF-beta(1)) has been reported previously. In this study, we investigated whether TGF-beta(1)-producing fibroblasts irradiated with a lethal dose of radiation are still capable of inducing the regeneration of hyaline articular cartilage. NIH 3T3TGF-beta(1) fibroblasts were exposed to doses of 20, 40, or 80 Gy, using a irradiator, and then injected into artificially made partial defects on the femoral condyle of rabbit knee joints. The rabbits were killed 3 or 6 weeks postinjection and hyaline articular cartilage regeneration was evaluated by histological and immunohistochemical staining (n = 5 per each group). Irradiated NIH 3T3-TGFbeta(1) fibroblasts started to die rapidly 3 days after irradiation; moreover, the kinetics of their viability were similar regardless of the radiation intensity. TGF-beta1 expression, measured by ELISA, showed that the TGF-beta(1) protein produced from the irradiated cells peaked 5 days after irradiation and thereafter declined rapidly. Complete filling of the defect with reparative tissue occurred in all the groups, although variations were observed in terms of the nature of the repair tissue. Histological and immunohistochemical staining of the repair tissue showed that the tissue newly formed by irradiated NIH 3T3-TGF-beta(1) fibroblasts after exposure to 20 Gy had hyaline cartilage-like characteristics, as was observed in the nonirradiated controls. On the other hand, the repair tissue formed by NIH 3T3-TGF-beta(1) fibroblasts irradiated with 40 or 80 Gy showed more fibrous cartilage-like tissue. These results suggest that TGF-beta(1)-producing fibroblasts irradiated up to a certain level of lethal dose (i.e., 20 Gy) are able to induce normal-appearing articular cartilage in vivo. Therefore, irradiated heterologous cell-mediated TGF-beta(1) gene therapy may be clinically useful and an efficient method of regenerating hyaline articular cartilage.

[1]  T. Takayama,et al.  Amelioration of murine dextran sulfate sodium‐induced colitis by ex vivo extracellular superoxide dismutase gene transfer , 2006, Inflammatory bowel diseases.

[2]  R. L. Reis,et al.  Biological response to pre-mineralized starch based scaffolds for bone tissue engineering , 2005, Journal of materials science. Materials in medicine.

[3]  S. Kim,et al.  Regeneration of hyaline cartilage by cell-mediated gene therapy using transforming growth factor beta 1-producing fibroblasts. , 2001, Human gene therapy.

[4]  Anders Lindahl,et al.  Autologous Chondrocyte Transplantation , 2002, The American journal of sports medicine.

[5]  B. Johnstone,et al.  Autologous mesenchymal progenitor cells in articular cartilage repair. , 1999, Clinical orthopaedics and related research.

[6]  S. O’Driscoll.  Current Concepts Review - The Healing and Regeneration of Articular Cartilage* , 1998 .

[7]  S D Gillogly,et al.  Treatment of articular cartilage defects of the knee with autologous chondrocyte implantation. , 1998, The Journal of orthopaedic and sports physical therapy.

[8]  P. D. Di Cesare,et al.  Chondrocyte transplantation and experimental treatment options for articular cartilage defects. , 1997, American journal of orthopedics.

[9]  E. Hunziker,et al.  Repair of Partial-Thickness Defects in Articular Cartilage: Cell Recruitment from the Synovial Membrane* , 1996, The Journal of bone and joint surgery. American volume.

[10]  C. Ohlsson,et al.  Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. , 1994, The New England journal of medicine.

[11]  S W O'Driscoll,et al.  Durability of regenerated articular cartilage produced by free autogenous periosteal grafts in major full-thickness defects in joint surfaces under the influence of continuous passive motion. A follow-up report at one year. , 1988, The Journal of bone and joint surgery. American volume.

[12]  D. Grande,et al.  Healing of experimentally produced lesions in articular cartilage following chondrocyte transplantation , 1987, The Anatomical record.

[13]  H. Mankin,et al.  The response of articular cartilage to mechanical injury. , 1982, The Journal of bone and joint surgery. American volume.

[14]  H J Mankin,et al.  The reaction of articular cartilage to injury and osteoarthritis (second of two parts). , 1974, The New England journal of medicine.

[15]  Mankin Hj,et al.  The reaction of articular cartilage to injury and osteoarthritis (first of two parts). , 1974 .

[16]  G. Bentley,et al.  Homotransplantation of Isolated Epiphyseal and Articular Cartilage Chondrocytes into Joint Surfaces of Rabbits , 1971, Nature.

[17]  P. Chesterman,et al.  Homotransplantation of articular cartilage and isolated chondrocytes. An experimental study in rabbits. , 1968, The Journal of bone and joint surgery. British volume.

[18]  H J Mankin,et al.  Articular cartilage: tissue design and chondrocyte-matrix interactions. , 1998, Instructional course lectures.

[19]  J. Mansour,et al.  Repair of large full-thickness articular cartilage defects with allograft articular chondrocytes embedded in a collagen gel. , 1998, Tissue engineering.

[20]  木村 友厚.  Chondrocytes embedded in collagen gels maintain cartilage phenotype during long-term cultures , 1985 .

[21]  G. Bentley,et al.  Isolated epiphyseal chondrocyte allografts into joint surfaces , 1978 .