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背景:骨质疏松是脊髓损伤的常见并发症之一,其确切机制尚未完全清楚,损伤平面上下骨质是否均发生骨质疏松,不同学者观点不同。目的:观察脊髓损伤后继发骨质疏松的骨组织超微结构、血液生化改变情况,分析损伤平面上下骨质受累情况和程度。设计:随机对照实验。单位:吉林大学第二临床医院动物试验室,吉林大学第二临床医院检验科,吉林大学白求恩医学部电镜教研室。材料:实验于2002-05/2003-05在吉林大学第二临床医院动物试验室完成。Wistar大鼠110只,均为雄性,四五月龄,体质量为300～320g。方法:110只Wistar雄性大鼠随机选取10只为0周空白对照组,将余下大鼠随机分为实验组和对照组,1,2,3,7,11周对照组及实验组每组各10只。用1mL/L戊巴比妥钠腹腔注射麻醉,对照组仅行T10椎板切除不破坏硬膜,不损伤脊髓,实验组大鼠于T10椎体水平切除椎板,行Allen’s法损伤脊髓。分别于0周、术后1周,2周,3周,7周,11周周末将动物下腔静脉采血4mL后麻醉状态下处死,血液离心,应用7170AHI-TACHI自动生化分析仪自动采样化验血钙、血磷的含量及碱性磷酸酶活性。另取0周空白对照组,7周实验组,11周实验组右侧肱骨、胫骨各一用25mL/L乙二胺四乙酸脱钙1个月后制备切片,在醋酸双氧铀及柠檬酸双重染色下应用JEM-1200型透射电子显微镜观察肱骨外科颈、胫骨平台部以骨细胞为主的超微结构改变情况。结果:进入结果分析的大鼠为105只。①血生化检查结果:实验2周时血磷含量对照组明显低于实验组犤(1.54±0.21),(2.76±0.16)mmol/L,(P<0.01)犦;实验3周时血钙含量实验组明显高于对照组犤(2.52±0.06),(2.35±0.12)mmol/L,(P<0.01)犦;实验7周时碱性磷酸酶活性实验组明显高于对照组犤(155.86±20.42),(129.25±7.30)Nμ/mg,(P<0.01)犦。②电子显微镜下观察结果:术后7周实验组胫骨标本可见骨细胞与骨陷窝分离,个别细胞胞核不规则,可见絮状物质、线立体粗面内质网肿胀开空化;11周时,可见骨细胞与骨陷窝严重分离。术后7周实验组肱骨标本可见骨细胞与骨陷窝分离,有的细胞核固缩粗面内质网肿胀空化;11周时仍见上述改变,但细胞肿胀空化程度减轻。结论:脊髓损伤早期破骨细胞活性增强,成骨细胞活性降低,导致骨吸收增强,骨形成减弱。鼠损伤平面上下骨质均受累,但不同部位的骨骼继发骨质疏松的程度不同,骨质疏松后骨细胞超微结构改变显著。
BACKGROUND: Osteoporosis is one of the common complications of spinal cord injury. The exact mechanism is not yet fully understood. Whether osteoporosis occurs at the upper and lower levels of the upper and lower levels of the lesion is different. OBJECTIVE: To observe the ultrastructures and blood biochemical changes of secondary osteoporosis after spinal cord injury, and analyze the status and degree of bone involvement in the upper and lower sides of the lesion. Design: Randomized controlled experiment. Unit: The Second Clinical Hospital of Jilin University Animal Laboratory, Jilin University Second Clinical Laboratory, Jilin University, Bethune Department of Medicine, Department of Electron Microscopy. Materials: The experiment was performed at Animal Laboratory of the Second Clinical Hospital of Jilin University from May 2002 to May 2003. 110 Wistar rats, all male, four or five months old, body weight of 300 ~ 320g. Methods: Totally 110 Wistar male rats were randomly divided into control group (n = 10) and control group (n = 10). The remaining rats were randomly divided into experimental group and control group 10 only Rats were anesthetized by intraperitoneal injection of 1 mL / L pentobarbital sodium. The control group only underwent T10 laminectomy without damaging the spinal cord. In the experimental group, the lamina were resected at the level of T10 vertebral body and Allen’s method was used to injure the spinal cord. Blood samples were taken from the inferior vena cava of 4 mL at 0 week, 1 week, 2 weeks, 3 weeks, 7 weeks and 11 weeks after the operation. The blood was centrifuged and the blood samples were taken automatically by 7170AHI-TACHI automatic biochemical analyzer Calcium, phosphorus content and alkaline phosphatase activity. Another 0 week blank control group, 7-week experimental group, 11-week experimental group right humerus, tibia with 25mL / L ethylene diamine tetraacetic acid decalcification 1 month after the preparation of uranyl acetate and citric acid Double staining under the application of JEM-1200 transmission electron microscopy of the humeral surgical neck, tibial plateau dominated by bone cell ultrastructural changes. Results: 105 rats entered the result analysis. ① blood biochemical test results: the experimental group 2 weeks when the phosphorus content of the control group was significantly lower than the experimental group 犤 (1.54 ± 0.21), (2.76 ± 0.16) mmol / L, (P <0.01) 犦; 3 weeks of experimental serum calcium The experimental group was significantly higher than that of the control group (2.52 ± 0.06) and (2.35 ± 0.12) mmol / L, respectively (P <0.01), and the activity of alkaline phosphatase in the experimental group was significantly higher than that of the control group 20.42), (129.25 ± 7.30) Nμ / mg, (P <0.01) 犦. ② Under electron microscope, the osteoblasts were separated from the lacunae in the experimental group at 7 weeks after operation. The nuclei of individual cells showed irregular nuclei. The flocculent material was seen. When seen, bone cells and bone lacuna serious separation. At 7 weeks after operation, the humerus specimens of the experimental group showed separation of osteocytes from the lacunae, swelling and cavitation of some nucleus pyknotic rough endoplasmic reticulum, and the above changes were still seen at 11 weeks, but the extent of cell cavitation cavitation was alleviated. Conclusion: The activity of osteoclasts increased at the early stage of spinal cord injury, osteoblast activity decreased, resulting in increased bone resorption and decreased bone formation. The level of bone in both the injured and the injured sides of rats was affected, but the degree of secondary osteoporosis was different in different parts of the bone. The ultrastructure of osteocytes in osteoporosis changed significantly.