Adhesion of transplanted chondrocytes onto cartilage in vitro and in vivo.
OBJECTIVE
The specific objectives of this study using organ culture were (1) to transplant chondrocytes onto an intact cartilage surface; (2) to genetically modify endogenous and transplanted chondrocytes; and (3) to assess the ability of these cells to continually express a gene product. The specific objective with in vivo experiments was to transplant chondrocytes with intraarticular injections to cartilage.


METHODS
Fluorescent membrane and intracellular dyes were used in conjunction with confocal microscopy to observe the integration of transplanted chondrocytes into cartilage both in vitro and in vivo. The distribution and duration of binding of rat, canine, and bovine chondrocytes to cartilage explants and the duration of expression of genes transduced into the transplanted chondrocytes were also determined. We used the vector AdlacZ, an E1 and E3 deleted replication defective adenoviral vector that contains the beta-galactosidase gene driven by the beta-actin promoter and the cytomegalovirus enhancer.


RESULTS
The transplanted chondrocytes had a patchy distribution after in vitro or in vivo transplantation and buried themselves within the cartilage over time. Chondrocytes infected with the adenoviral vector AdlacZ soon or well after transplant to cartilage explants were maintained on the cartilage and continued throughout the duration of each trial to produce beta-galactosidase coded by the adenoviral vector. The cartilage plugs were infected with AdlacZ at 2 days or one, 2, 5, or 8 weeks after the chondrocytes were transplanted. The cartilage slices were then cultured from 15 days for chondrocytes infected at 8 weeks to 60 days for chondrocytes infected at 2 days post-transplant before determining the expression of beta-galactosidase.


CONCLUSIONS
These results support the possibility of repairing cartilage by intraarticular injections of chondrocytes. Transduction of chondrocytes with genes producing a variety of matrix promoting proteins should further enhance the reconstruction of osteoarthritic cartilage.
Doherty
P J
PJ
Department of Medicine, The Toronto Hospital, Ontario, Canada.
Zhang
H
H
Manolopoulos
V
V
Trogadis
J
J
Tremblay
L
L
Marshall
K W
KW
D000818:Animals; D002232:Carbocyanines; D002356:Cartilage / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D002417:Cattle; D019902:Chondrocytes / Q000637:transplantation*; D004285:Dogs; D005456:Fluorescent Dyes; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D007270:Injections, Intra-Articular; D007763:Lac Operon / Q000235:genetics; D010003:Osteoarthritis / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D014162:Transfection
C035005:3,3'-dihexadecylindocarbocyanine; D002232:Carbocyanines; D005456:Fluorescent Dyes