Rat Intervertebral Disc Cells. AAV-hTGF ¦ÂI Vitro Transfection Re-transplantation Experiments

ObjectiveTo observe the effect of AAV-hTGFβ1 transfected cells transplantat-ion on height of intervertebral disc, collagen typeⅡand proteoglycan of rat.MethodsOne hundred of intervertebral discs from forty rats were evenly divided into three groups:blank control group of forty L6/S1 normal discs without treatment, puncture group of forty L5/L6 discs were treated with 0.01 mol/L PBS injection, experimental group of forty L4/L5 discs were injected with AAV-hTGFβ1 transfected disc cells. All the rats received the radiographic study before the operation. Twenty rats received the radiograp-hic study respectively 4 weeks and 8 weeks after operation, then sacrificed and subjected to Semi-quantitative reverse transcription polymerase chain reaction to measure collagen type II and proteoglycan content.Results4 weeks after operation, the %DHIs of puncture group were remarkab-ly decreased compared with experimental group and blank control group (F =321.22,q=30.44,32.56,p<0.05).there was no statistical difference between experimental group and blank control group (P>0.05). Collagen typeⅡand proteoglycan gene expression in puncture group was weaker than in experim-ental group and blank control group (F=55.05.34.79, q=8.96～12.43, P <0.05); there was no statistical difference between experimental group and blank control group (P>0.05).8 weeks after operation, the %DHIs of punctu-re group were remarkably decreased compared with experimental group and blank control group (F=376.28,q=22.93,24.49,P< 0.05).there was no stati stical difference between experimental group and blank control group (P>0. 05). Collagen type II and proteoglycan gene expression in puncture group was weaker than in experim-ental group and blank control group (F=53.42,32.56, q=6.49-11.37,P<0.05); there was no statistical difference between experimental group and blank control group (P>0.05).Conclusion1. The AAV vector efficiently transduced intervertebral disc cells of rats and provided a strong transgene expression.2. Our research also confirm that the cell transplantation of disc cells is feasible on the rats.3. Transplantation of AAV-hTGFβ1 transfected disc cells into the intervertebral discs could enhance the expression of typeⅡ collagen and proteoglycan.4. Transplantation of AAV-hTGFβ1 transfected disc cells into the intervertebral discs could contribute to the maintenance of intervertebral height...