In vivo gene delivery to articular chondrocytes mediated by an adeno-associated virus vector.
OBJECTIVE
(1) To investigate the efficiency of direct in vivo adeno-associated virus (AAV) vector-mediated gene transduction to chondrocytes in relation to normal and injured articular cartilage. (2) To evaluate the effects of ultra-violet light-activated gene transduction (LAGT) in chondrocytes in vivo. (3) To determine dissemination of active rAAV vector after intra-articular administration.


METHODS
Rabbit knees with either normal or injured cartilage received an intra-articular injection with 1.5x10(12) infectious rAAV-eGFP particles. The right knees received rAAV-eGFP alone, whereas the left knees were given LAGT-treatment. The transduction efficiencies were determined at 1 and 3 weeks after infection by fluorescence-activated cell scanning. The occurrence of active shedding was monitored in serum and various tissues.


RESULTS
After 1 week, 7% of the chondrocytes in normal cartilage were transduced by direct rAAV transduction technique. Chondrocytes in cartilage defects demonstrated higher transduction rates compared to chondrocytes in normal cartilage. LAGT increased the cellular eGFP expression in the internal zones to 12%, but did not have any effect in the external zones in defects. Finally, infectious particles were not detected in either serum or tissue samples.


CONCLUSIONS
Direct rAAV-mediated gene transfer in vivo to articular chondrocytes is possible. LAGT improves rAAV transduction of chondrocytes in vivo but appears to have a very limited range of effect induction. Expression of eGFP was not determined in other tissues than synovium and cartilage in the treated joints.
Ulrich-Vinther
M
M
Department of Molecular Biology, University of Aarhus, Aarhus, Denmark.
Duch
M R
MR
Søballe
K
K
O'Keefe
R J
RJ
Schwarz
E M
EM
Pedersen
F S
FS
D000818:Animals; D017209:Apoptosis / Q000528:radiation effects; D002358:Cartilage, Articular / Q000293:injuries / Q000378:metabolism* / Q000473:pathology; D002470:Cell Survival / Q000528:radiation effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism* / Q000473:pathology / Q000528:radiation effects; D000229:Dependovirus / Q000235:genetics*; D005260:Female; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006614:Hindlimb; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D013583:Synovial Membrane / Q000378:metabolism / Q000473:pathology / Q000528:radiation effects; D014161:Transduction, Genetic
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins