TY - JOUR
T1 - Enhanced repair of the anterior cruciate ligament by in situ gene transfer
T2 - Evaluation in an in vitro model
AU - Pascher, Arnulf
AU - Steinert, Andre F.
AU - Palmer, Glyn D.
AU - Betz, Oliver
AU - Gouze, Jean Noel
AU - Gouze, Elvire
AU - Pilapil, Carmencita
AU - Ghivizzani, Stephen C.
AU - Evans, Christopher H.
AU - Murray, Martha Meaney
N1 - Funding Information:
A.P. was supported by a postdoctoral fellowship from the Fond zur Foerderung der Wissenschaftlichen Forschung, Austria, and A.F.S. was supported by a postdoctoral fellowship from the Deutsche Forschungsgemeinschaft, Germany, Grant STE-1051/1-1. Additional support was provided by the National Institute of Arthritis and Musculoskeletal and Skin Diseases, a part of the National Institutes of Health, Grant AR 46356 (M.M.M.), and by the National Football League Medical Charities Grant Program (M.M.M.).
PY - 2004/8
Y1 - 2004/8
N2 - The inability of the ruptured anterior cruciate ligament (ACL) of the knee joint to heal spontaneously presents numerous clinical problems. Here we describe a novel, gene-based approach to augment ACL healing. It is based upon the migration of cells from the ruptured ends of the ligament into a collagen hydrogel laden with recombinant adenovirus. Cells entering the gel become transduced by the vector, which provides a basis for the local synthesis of gene products that aid repair. Monolayers of bovine ACL cells were readily transduced by first-generation, recombinant adenovirus, and transgene expression remained high after the cells were incorporated into collagen hydrogels. Using an in vitro model of ligament repair, cells migrated from the cut ends of the ACL into the hydrogel and were readily transduced by recombinant adenovirus contained within it. The results of experiments in which GFP was used as the transgene suggest highly efficient transduction of ACL cells in this manner. Moreover, during a 21-day period GFP+ cells were observed more than 6 mm from the severed ligament. This distance is ample for the projected clinical application of this technology. In response to TGF-β1 as the transgene, greater numbers of ACL cells accumulated in the hydrogels, where they deposited larger amounts of type III collagen. These data confirm that it is possible to transduce ACL cells efficiently in situ as they migrate from the ruptured ACL, that transduction does not interfere with the cells' ability to migrate distances necessary for successful repair, and that ACL cells will respond in a suitable manner to the products of the transgenes they express. This permits optimism over a possible clinical use for this technology.
AB - The inability of the ruptured anterior cruciate ligament (ACL) of the knee joint to heal spontaneously presents numerous clinical problems. Here we describe a novel, gene-based approach to augment ACL healing. It is based upon the migration of cells from the ruptured ends of the ligament into a collagen hydrogel laden with recombinant adenovirus. Cells entering the gel become transduced by the vector, which provides a basis for the local synthesis of gene products that aid repair. Monolayers of bovine ACL cells were readily transduced by first-generation, recombinant adenovirus, and transgene expression remained high after the cells were incorporated into collagen hydrogels. Using an in vitro model of ligament repair, cells migrated from the cut ends of the ACL into the hydrogel and were readily transduced by recombinant adenovirus contained within it. The results of experiments in which GFP was used as the transgene suggest highly efficient transduction of ACL cells in this manner. Moreover, during a 21-day period GFP+ cells were observed more than 6 mm from the severed ligament. This distance is ample for the projected clinical application of this technology. In response to TGF-β1 as the transgene, greater numbers of ACL cells accumulated in the hydrogels, where they deposited larger amounts of type III collagen. These data confirm that it is possible to transduce ACL cells efficiently in situ as they migrate from the ruptured ACL, that transduction does not interfere with the cells' ability to migrate distances necessary for successful repair, and that ACL cells will respond in a suitable manner to the products of the transgenes they express. This permits optimism over a possible clinical use for this technology.
KW - Anterior cruciate ligament
KW - Collagen hydrogel
KW - Gene therapy
KW - TGF-β
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U2 - 10.1016/j.ymthe.2004.03.012
DO - 10.1016/j.ymthe.2004.03.012
M3 - Article
C2 - 15294179
AN - SCOPUS:4344595112
SN - 1525-0016
VL - 10
SP - 327
EP - 336
JO - Molecular Therapy
JF - Molecular Therapy
IS - 2
ER -