title: Overexpression of TGF-β via rAAV-Mediated Gene Transfer Promotes the Healing of Human Meniscal Lesions Ex Vivo on Explanted Menisci.
abstract: BACKGROUND
  Direct application of therapeutic gene vectors derived from the adeno-associated virus (AAV) might be beneficial to improve the healing of meniscal tears.
  
  
  OBJECTIVE
  To test the ability of recombinant AAV (rAAV) to overexpress the potent transforming growth factor-β (TGF-β) in primary cultures of human meniscal fibrochondrocytes, in human meniscal explants, and in experimental human meniscal lesions as a new tool to enhance meniscal repair.
  
  
  METHODS
  Controlled laboratory study.
  
  
  METHODS
  The effects of the candidate treatment on the proliferative and metabolic activities of meniscal cells were monitored in vitro for up to 21 days and in situ in intact and injured human menisci for up to 15 days using biochemical, immunohistochemical, histological, and histomorphometric analyses.
  
  
  RESULTS
  Efficient production of TGF-β via rAAV was achieved in vitro and in situ, both in the intact and injured meniscus. Application of the rAAV TGF-β vector stimulated the levels of cell proliferation and matrix synthesis (type I collagen) compared with control gene transfer in all systems tested, especially in situ in regions of poor healing capacity and in sites of meniscal injury. No adverse effects of the candidate treatment were observed at the level of osseous differentiation, as tested by immunodetection of type X collagen. Most remarkably, a significant reduction of the amplitude of meniscal tears was noted after TGF-β treatment, an effect that was associated with increased expression levels of the α-smooth muscle actin contractile marker.
  
  
  CONCLUSIONS
  Overexpression of TGF-β via rAAV gene transfer is capable of modulating the reparative activities of human meniscal cells, allowing for the healing of meniscal lesions by convenient injection in sites of injury.
  
  
  CONCLUSIONS
  Direct gene-based approaches using rAAV have strong potential to develop new therapeutic options that aim at treating human meniscal defects.
authors: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany mmcucchiarini@hotmail.com.
authors: Magali
authors: M
authors: Cucchiarini
authors: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany.
authors: Katharina
authors: K
authors: Schmidt
authors: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany.
authors: Janina
authors: J
authors: Frisch
authors: Department of Orthopaedic Surgery, Saarland University Medical Center, Homburg/Saar, Germany.
authors: Dieter
authors: D
authors: Kohn
authors: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany Department of Orthopaedic Surgery, Saarland University Medical Center, Homburg/Saar, Germany.
authors: Henning
authors: H
authors: Madry
mesh_terms: D000368:Aged; D002357:Cartilage Diseases / Q000235:genetics / Q000628:therapy*; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007718:Knee Injuries / Q000235:genetics / Q000628:therapy*; D008875:Middle Aged; D016212:Transforming Growth Factor beta / Q000235:genetics*; D014945:Wound Healing / Q000235:genetics
chemical_list: D016212:Transforming Growth Factor beta
keywords: TGF-β; gene therapy; human meniscus; meniscal lesions; rAAV