Direct in vivo gene transfer to healing rat patellar ligament by intra-arterial delivery of haemagglutinating virus of Japan liposomes.
BACKGROUND
Manipulation of ligament healing has been a major focus of orthopaedic research. In recent years, gene transfer to healing ligament appears to be a feasible method for manipulating the healing process. In this study, we investigated the feasibility of gene transfer to healing rat patellar ligament by intra-arterial delivery.


METHODS
An attempt was made to transfer a reporter gene (Escherichia coli, beta-galactosidase gene) to healing rat patellar ligament using the haemagglutinating virus of Japan (HVJ) liposome-mediated gene transfer method. Three days after cutting the patellar tendons of 25 14-week-old male Wistar rats, HVJ-liposome complexes containing beta-galactosidase (beta-gal) cDNA were injected into the femoral artery of 15 Wistar rats as the experimental group. HVJ liposomes without DNA were injected into the femoral artery of 10 Wistar rats as the control group. Three rats from the experimental group and two control rats were killed 3, 7, 14, 28 and 56 days after the injection.


RESULTS
After X-gal staining, the rate of transfection in the experimental group (mean +/- SEM) was found to be 12.1% +/- 0.590%, 8.7% +/- 0.217%, 10.2% +/- 0.227%, 3.2% +/- 0.247% and 0.7% +/- 0.060% at post-injection days 3, 7, 14, 28 and 56 respectively. In control sections the number of blue-stained cells were very few at any point.


CONCLUSIONS
We succeeded in introducing a reporter gene into healing rat patellar ligament by infra-arterial delivery of HVJ-liposome complexes. This method appears to have the potential to be applicable for soft-tissue healing studies and also healing studies of other tissues and organs.
Ozkan
I
I
Osaka University Medical School, Japan. iozkan@superonline.com
Shino
K
K
Nakamura
N
N
Natsuume
T
T
Matsumoto
N
N
Horibe
S
S
Tomita
T
T
Kaneda
Y
Y
Ochi
T
T
D000818:Animals; D005263:Femoral Artery; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D007269:Injections, Intra-Arterial; D008081:Liposomes / Q000008:administration & dosage; D008297:Male; D017847:Patellar Ligament / Q000293:injuries*; D051381:Rats; D017208:Rats, Wistar; D018108:Respirovirus / Q000235:genetics*; D014162:Transfection; D014945:Wound Healing; D001616:beta-Galactosidase / Q000235:genetics
D008081:Liposomes; D001616:beta-Galactosidase