ACUTE MECHANICAL INJURY OF THE HUMAN INTERVERTEBRAL DISC: LINK TO DEGENERATION AND PAIN
ACUTE MECHANICAL INJURY OF THE HUMAN INTERVERTEBRAL DISC: LINK TO DEGENERATION AND PAIN
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DOI：
10.22203/ecm.v028a08
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发表时间：
2014-07-01
影响因子：
3.1
通讯作者：
Haglund, L.
中科院分区：
文献类型：
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作者：
Alkhatib, B.;Rosenzweig, D. H.;Haglund, L.
过度的机械负荷或椎间盘（IVD）的急性创伤被认为是导致退变和疼痛的原因。然而，机械损伤引发和促进变性的确切机制仍不清楚。本研究探讨了急性机械损伤后全器官人IVD培养物中的生化变化和细胞外基质破坏。将分离的健康人IVD快速压缩椎间盘高度的5%（未受伤）或30%（受伤）。30%的应变持续导致软骨终板破裂，证实椎间盘损伤。加载后三天，评估条件培养基的蛋白聚糖含量和释放的细胞因子。评估组织提取物的蛋白聚糖含量和聚集蛋白聚糖完整性。将条件培养基应用于PC 12细胞以评估诱导神经突生长的因子是否被释放。与对照组相比，IVD损伤导致显著的细胞死亡。损伤还导致组织蛋白多糖含量显著降低，培养基中蛋白多糖含量相应增加。由于基质金属蛋白酶和聚集蛋白聚糖酶活性增加，在损伤组织中观察到聚集蛋白聚糖片段化增加。损伤的IVD条件培养基含有显著升高的白细胞介素（IL）-5、IL-6、IL-7、IL-8、MCP-2、GRO α和TNF α，并且ELISA分析显示，与未损伤的培养基相比，神经生长因子水平显著升高。与未损伤的培养基相比，损伤的椎间盘培养基在PC 12细胞中引起显著的神经突发芽。离体人IVD的急性机械损伤引发与变性和背痛相关的因子和酶活性的释放。这项工作提供了直接证据，将体内急性创伤、炎症因子、新神经支配和潜在的变性和椎间盘源性疼痛联系起来。
Excessive mechanical loading or acute trauma to intervertebral discs (IVDs) is thought to contribute to degeneration and pain. However, the exact mechanisms by which mechanical injury initiates and promotes degeneration remain unclear. This study investigates biochemical changes and extracellular matrix disruption in whole-organ human IVD cultures following acute mechanical injury. Isolated healthy human IVDs were rapidly compressed by 5% (non-injured) or 30% (injured) of disc height. 30% strain consistently cracked cartilage endplates, confirming disc trauma. Three days post-loading, conditioned media were assessed for proteoglycan content and released cytokines. Tissue extracts were assessed for proteoglycan content and for aggrecan integrity. Conditioned media were applied to PC12 cells to evaluate if factors inducing neurite growth were released. Compared to controls, IVD injury caused significant cell death. Injury also caused significantly reduced tissue proteoglycan content with a reciprocal increase of proteoglycan content in culture media. Increased aggrecan fragmentation was observed in injured tissue due to increased matrix metalloproteinase and aggrecanase activity. Injured-IVD conditioned media contained significantly elevated interleukin (IL)-5, IL-6, IL-7, IL-8, MCP-2, GRO alpha, and MIG, and ELISA analysis showed significantly increased nerve growth factor levels compared to non-injured media. Injured-disc media caused significant neurite sprouting in PC12 cells compared to non-injured media. Acute mechanical injury of human IVDs ex vivo initiates release of factors and enzyme activity associated with degeneration and back pain. This work provides direct evidence linking acute trauma, inflammatory factors, neo-innervation and potential degeneration and discogenic pain in vivo.