Wehling P, Schulitz K P, Robbins P D, Evans C H, Reinecke J A
Praxis und Klinik für Orthopädie und Neurochirurgie, Düsseldorf, Germany.
Spine (Phila Pa 1976). 1997 May 15;22(10):1092-7. doi: 10.1097/00007632-199705150-00008.
In the current study, chondrocytic cells from bovine intervertebral end plates were cultivated in vitro and modified genetically.
The authors intended to perform isolation and cultivation of cells from bovine end plates of the spine. They also intended to show, in principle, the feasibility of introducing exogenous genes into chondrocytic cells from bovine intervertebral end plates by way of retroviral vectors.
The involvement of cytokines in the destruction of articular cartilage is established. It appears possible that similar mechanisms may play a role in intervertebral disc degeneration and other spinal disorders. Conventional medication and surgery of intervertebral disc degeneration addresses neither the pathophysiology nor the chronicity of the disease. Therapeutic proteins carry great potential as locally produced drugs after transfer of their cognate genes to the sites of interest.
Vertebral end plate tissue was obtained from bovine os coccygis. Chondrocytic cells were isolated and cultured in vitro. The bacterial beta-galactosidase (LacZ) gene and, alternatively, the complementary DNA (DNA copy of the mRNA) of the human interleukin-1 receptor antagonist were introduced into the isolated cells by retrovirus mediated gene transfer. beta-galactosidase activity was determined by staining, and interleukin-1 receptor antagonist protein was quantitated by enzyme-linked immunosorbent assay.
Isolation and cultivation of chondrocytic end plate cells is possible. Native cells continue to grow in culture for more than 2 months. Transfer of the beta-galactosidase gene to cultured cells resulted in approximately 1% beta-galactosidase positive cells. Transfer of the interleukin-1 receptor antagonist complementary DNA resulted in the production of 24 ng/ml/10(6) cells interleukin-1 receptor antagonist protein in 48 hours.
The introduction of exogenous therapeutic genes into cells from the intervertebral end plate opens the possibility for a local gene-based treatment of intervertebral disc degeneration. This approach avoids some of the shortcomings of conventional drug- and surgery-based treatments and has the potential to be specific, effective, and appropriate to the chronicity of the disease.
在本研究中，对来自牛椎间终板的软骨细胞进行体外培养并进行基因改造。
作者旨在从牛脊柱终板中分离和培养细胞。他们还打算原则上证明通过逆转录病毒载体将外源基因导入牛椎间终板软骨细胞的可行性。
细胞因子参与关节软骨破坏已得到证实。类似的机制可能在椎间盘退变和其他脊柱疾病中起作用。椎间盘退变的传统药物治疗和手术治疗既不能解决疾病的病理生理学问题，也不能解决其慢性问题。治疗性蛋白质在将其同源基因转移到靶位点后作为局部产生的药物具有巨大潜力。
从牛尾骨获取椎体终板组织。分离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）基因以及人白细胞介素-1受体拮抗剂的互补DNA（mRNA的DNA拷贝）导入分离的细胞。通过染色测定β-半乳糖苷酶活性，通过酶联免疫吸附测定法定量白细胞介素-1受体拮抗剂蛋白。
可以分离和培养软骨终板细胞。原代细胞在培养中持续生长超过2个月。将β-半乳糖苷酶基因转移到培养细胞中导致约1%的β-半乳糖苷酶阳性细胞。转移白细胞介素-1受体拮抗剂互补DNA导致在48小时内每10⁶个细胞产生24 ng/ml的白细胞介素-1受体拮抗剂蛋白。
将外源治疗基因导入椎间终板细胞为基于局部基因的椎间盘退变治疗开辟了可能性。这种方法避免了传统药物和手术治疗的一些缺点，并且有可能具有特异性、有效性且适合疾病的慢性特点。
