Repair of articular cartilage defect by intraarticular administration of basic fibroblast growth factor gene, using adeno-associated virus vector.
The objective of this study was to establish the potency of adeno-associated virus (AAV) as a viral vector to transport the basic fibroblast growth factor (bFGF) gene into synovial tissue, and to evaluate the consequent repair of articular cartilage defects. In the in vitro study, LacZ- and bFGF-encoding genes were transduced into rabbit synoviocytes by recombinant adeno-associated virus (AAV) vector, and the cells were cultured for 2 weeks. The percentage of successfully transduced LacZ-positive cells was assessed by 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside staining, and the concentration of bFGF in the culture supernatant was confirmed by bFGF-specific enzyme-linked immunosorbent assay. In the in vivo study, 12- to 14-week-old Japanese white rabbits (all female) were used. AAV-bFGF was administered into an artificially created full-thickness defect (5 mm in diameter and 3 mm deep) in the patellar groove of the distal femur. Cartilage repair was subsequently monitored at 4, 8, and 12 weeks, by macroscopic and histological examination, and results were graded on the basis of semiquantitative scores. lacZ gene expression in synoviocytes reached more than 93% within the first 2 weeks, and the mean bFGF concentration in the culture supernatant of the bFGF gene-transduced group was significantly increased (p < 0.01). Semiquantitative macroscopic and histological assessment indicated that the average score was significantly better in the bFGF-transduced group throughout the observation period, suggesting better cartilage repair. These results demonstrate that gene transfer into synoviocytes, using the AAV vector, was a potent method of gene transduction. Moreover, after intraarticular administration of AAV-bFGF, constant expression of bFGF in the knee joints resulted in substantial cartilage regeneration that, with further long-term study, could possibly merit consideration for clinical application.
Hiraide
Atsuo
A
Department of Orthopedic Surgery, Yokohama City University School of Medicine, Yokohama, Japan. arzt2001@h9.dion.ne.jp
Yokoo
Naoki
N
Xin
Ke-Qin
KQ
Okuda
Kenji
K
Mizukami
Hiroaki
H
Ozawa
Keiya
K
Saito
Tomoyuki
T
D000818:Animals; D002358:Cartilage, Articular / Q000187:drug effects* / Q000293:injuries / Q000502:physiology; D020219:Chondrogenesis; D000229:Dependovirus / Q000235:genetics*; D016326:Extracellular Matrix Proteins; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000235:genetics* / Q000494:pharmacology; D015316:Genetic Therapy; D005822:Genetic Vectors; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007763:Lac Operon / Q000235:genetics*; D011817:Rabbits; D012038:Regeneration; D013582:Synovial Fluid / Q000166:cytology; D014161:Transduction, Genetic
D016326:Extracellular Matrix Proteins; D016222:Fibroblast Growth Factor 2