Reinecke J A, Wehling P, Robbins P, Evans C H, Sager M, Schulze-Allen G, Koch H
Orthogen Gentechnologie GmbH, Düsseldorf.
Z Orthop Ihre Grenzgeb. 1997 Sep-Oct;135(5):412-6. doi: 10.1055/s-2008-1039409.
It is well known that cytokines are involved in the induction of intervertebral disc and articular cartilage destruction. Therapeutic proteins are of great potential as locally produced drugs after transfer of their cognate genes to the sites of interest.
Chondrocytic cells from bovine os coccygis and discal chondrocytes from 6 wistar rats were isolated and cultured in vitro. The bacterial beta-galactosidase (LacZ) gene and the cDNA of the human interleukin-1 receptor antagonist (IL-1ra) were introduced into the cells by retrovirus mediated gene transfer. LacZ activity was determined by Xgal staining, IL-1ra protein was determined by ELISA.
Our study confirms that isolation and cultivation of bovine chondrocytic end plate cells and of rat discal cells in possible. Transfer of both LacZ and of the IL-1ra cDNA to cultured cells was successfull.
The introduction of exogenous therapeutical genes into cells from the intervertebral disc and the end plate opens the possibility for a local gene therapy of IVD degeneration. This therapy has the potential to be specific, effective and appropriate to the chronicity of the disease.
众所周知，细胞因子参与椎间盘和关节软骨破坏的诱导过程。治疗性蛋白质在将其同源基因转移至靶位点后作为局部产生的药物具有巨大潜力。
从牛尾骨分离软骨细胞以及从6只Wistar大鼠分离椎间盘软骨细胞，并进行体外培养。通过逆转录病毒介导的基因转移将细菌β-半乳糖苷酶（LacZ）基因和人白细胞介素-1受体拮抗剂（IL-1ra）的cDNA导入细胞。通过Xgal染色测定LacZ活性，通过ELISA测定IL-1ra蛋白。
我们的研究证实了牛软骨终板细胞和大鼠椎间盘细胞的分离与培养是可行的。LacZ和IL-1ra cDNA成功转移至培养细胞。
将外源性治疗基因导入椎间盘和终板细胞为椎间盘退变的局部基因治疗开辟了可能性。这种治疗具有针对疾病、有效且适合疾病慢性特征的潜力。
