Effects of growth differentiation factor-5 on the intervertebral disc -: In vitro bovine study and in vivo rabbit disc degeneration model study
Effects of growth differentiation factor-5 on the intervertebral disc -: In vitro bovine study and in vivo rabbit disc degeneration model study
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DOI：
10.1097/01.brs.0000248428.22823.86
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发表时间：
2006-12-01
期刊：
影响因子：
3
通讯作者：
Masuda, Koichi
中科院分区：
文献类型：
--
作者：
Chujo, Takehide;An, Howard S.;Masuda, Koichi
研究设计。重组人生长与分化因子-5 （rhGDF-5）对牛椎间盘细胞基质代谢影响的体外研究及rhGDF-5对兔环形穿刺模型的体内影响研究。测定rhGDF-5对椎间盘的修复能力。背景资料摘要。rhGDF-5是发育中的肌肉骨骼系统中的一种重要蛋白，其在体外和体内对退变椎间盘的修复作用尚不清楚。体外分别用rhGDF-5（100或200 ng/mL）培养牛髓核和纤维环细胞。测定第7、14、21天的脱氧核糖核酸和蛋白聚糖含量，以及蛋白聚糖和胶原蛋白的合成。在体内，16只青春期新西兰大白兔在2个腰椎间盘穿刺。4周后，向髓核注射磷酸缓冲盐水或rhGDF-5 （10 ng、1和100 μ g）。随访16周，观察椎间盘高度、磁共振成像及组织学分级。体外培养14 d后，rhGDF-5显著提高了两种细胞的脱氧核糖核酸和蛋白聚糖含量。200 ng/mL rhGDF-5显著刺激第21天蛋白多糖合成（髓核+138%，纤维环+24%）和胶原合成（髓核+95%，纤维环+23%）。体内注射rhGDF-5后椎间盘高度恢复，磁共振成像评分和组织学分级评分提高，差异均有统计学意义（P < 0.05 ~ 0.001）。单次注射rhGDF-5对椎间盘具有修复能力，可能是基于其在体外增强细胞外基质生成的作用。
Study Design. In vitro studies on the effects of recombinant human growth and differentiation factor-5 (rhGDF-5) on matrix metabolism of bovine intervertebral disc cells and an in vivo study on the effect of rhGDF-5 in the rabbit anular puncture model.Objective. To determine the reparative capacity of rhGDF-5 on the intervertebral disc.Summary of Background Data. The in vitro and in vivo effects of rhGDF-5, a crucial protein in the developing musculoskeletal system, on repair of the degenerated intervertebral disc remain unidentified.Methods. In vitro, bovine nucleus pulposus and anulus fibrosus cells were cultured with or without rhGDF-5 (100 or 200 ng/mL). On days 7, 14, and 21, the contents of deoxyribonucleic acid and proteoglycan, and the synthesis of proteoglycan and collagen were assessed. In vivo, 16 adolescent New Zealand white rabbits received anular punctures in 2 lumbar discs. Four weeks later, phosphate buffered saline or rhGDF-5 (10 ng, 1 and 100 mu g) was injected into the nucleus pulposus. The rabbits were followed up for 16 weeks for disc height, magnetic resonance imaging, and histologic grading.Results. In vitro, rhGDF-5 increased the deoxyribonucleic acid and proteoglycan contents of both cell types significantly after day 14. rhGDF-5 at 200 ng/mL significantly stimulated proteoglycan synthesis (nucleus pulposus: +138%, anulus fibrosus: +24%) and collagen synthesis (nucleus pulposus: +95%, anulus fibrosus: +23%) at day 21. In vivo, the injection of rhGDF-5 resulted in a restoration of disc height, improvement of magnetic resonance imaging scores, and histologic grading scores with statistical significance (P < 0.05 - 0.001).Conclusion. A single injection of rhGDF-5 has a reparative capacity on intervertebral discs, presumably based on its effects to enhance extracellular matrix production in vitro.