Haro H, Murakami S, Komori H, Okawa A, Shinomiya K
Department of Orthopaedic Surgery, Faculty of Medicine, Tokyo Medical and Dental University.
Spine (Phila Pa 1976). 1997 May 15;22(10):1098-104. doi: 10.1097/00007632-199705150-00009.
Immunohistologic analysis was performed on surgically removed samples of herniated nucleus pulposus to examine the expression of stromelysin-1. We performed in vitro and in vivo experiments to determine whether recombinant human (rh) stromelysin-1 is capable of degrading nucleus pulposus.
To analyze the production of stromelysin-1 in various types of herniated nucleus pulposus, and to examine the effects of this recombinant protein on nucleus pulposus tissues.
The authors previously demonstrated a progressive decrease in herniated nucleus pulposus size in some of the transligamentous and sequestration types of herniated nucleus pulposus using magnetic resonance imaging. An increased production of stromelysin-1, a cartilage proteoglycan degrading enzyme, in herniated nucleus pulposus was reported recently. The authors speculated that if stromelysin-1 is involved in the degradation of herniated nucleus pulposus, stromelysin-1 itself may be used as a chemonucleolytic agent.
Immunohistologic analysis using streptoavidin-biotin method was performed on 20 herniated nucleus pulposus samples to investigate the expression of stromelysin-1. Five herniated nucleus pulposus samples were incubated in a tissue culture medium in the presence or absence of rh stromelysin-1. After 24 hours of incubation, their weight changes were measured, and the loss of proteoglycan was assessed by Safranin O staining. Rat nucleus pulposus tissues were obtained from coccygeal intervertebral discs, and autologous subcutaneous transplantation was performed. Rh stromelysin-1 was injected into the grafted materials, and the reduction in size was followed by two-dimensional measurements from the skin surface, using engineer's calipers.
Immunohistologic analysis demonstrated the production of stromelysin-1 in the granulation tissues of herniated nucleus pulposus. When stromelysin-1 was injected into the murine nucleus pulposus tissues, they reduced in size more rapidly than the control group. In addition, human herniated nucleus pulposus materials obtained at surgery showed significant weight loss when treated with stromelysin-1 in an organ culture system. Safranin O staining revealed extensive depletion of proteoglycan in these herniated nucleus pulposus samples.
Stromelysin-1 is a possible key enzyme in herniated nucleus pulposus resorption, and stromelysin-1 may be a good candidate for use in chemonucleolysis. Administration of human stromelysin-1 may physiologically facilitate the resorption process of herniated nucleus pulposus, increase the healing rate and decrease complications after chemonucleolysis.
对手术切除的髓核突出样本进行免疫组织学分析，以检测基质溶解素-1的表达。我们进行了体外和体内实验，以确定重组人基质溶解素-1是否能够降解髓核。
分析不同类型髓核突出中基质溶解素-1的产生情况，并研究这种重组蛋白对髓核组织的影响。
作者先前使用磁共振成像证明，在一些经韧带型和游离型髓核突出中，髓核突出的大小会逐渐减小。最近有报道称，髓核突出中软骨蛋白聚糖降解酶基质溶解素-1的产生增加。作者推测，如果基质溶解素-1参与了髓核突出的降解，那么基质溶解素-1本身可能可作为一种化学溶核剂。
对20个髓核突出样本采用链霉抗生物素蛋白-生物素方法进行免疫组织学分析，以研究基质溶解素-1的表达。将5个髓核突出样本在有或无重组人基质溶解素-1的情况下置于组织培养基中孵育。孵育24小时后，测量其重量变化，并通过番红O染色评估蛋白聚糖的丢失情况。从尾椎椎间盘获取大鼠髓核组织，并进行自体皮下移植。将重组人基质溶解素-1注入移植材料中，使用游标卡尺从皮肤表面进行二维测量，跟踪其大小的减小情况。
免疫组织学分析表明，髓核突出的肉芽组织中产生了基质溶解素-1。当将基质溶解素-1注入小鼠髓核组织时，它们的大小比对照组减小得更快。此外，在器官培养系统中用基质溶解素-1处理手术获取的人髓核突出材料时，其重量显著减轻。番红O染色显示这些髓核突出样本中的蛋白聚糖大量减少。
基质溶解素-1可能是髓核突出吸收的关键酶，并且基质溶解素-1可能是化学溶核的良好候选药物。给予人基质溶解素-1可能在生理上促进髓核突出的吸收过程，提高愈合率并减少化学溶核后的并发症。
