新鲜羊膜对周围神经损伤修复影响的实验研究

发布时间：2018-08-08 17:55

【摘要】：目的:周围神经损伤是临床上较常见的致残性疾病,直接损害了劳动力资源,给社会和家庭生活造成沉重的负担。周围神经损伤修复后再生受多种因素的影响,吻合口处的微环境及局部的癜痕形成严重影响了神经损伤后修复效果,为此,改善局部微环境及减少局部瘢痕有利于周围神经再生促进神经功能恢复。羊膜具有半透性,保证营养物质顺利透过,促进雪旺细胞增殖,加速神经轴索生长,抑制周围成纤维细胞的生物活性,限制神经周围组织侵入神经断端,抑制周围瘢痕形成,防止神经粘连及神经压迫等。本实验构建大鼠坐骨神经损伤模型,在显微镜下行神经吻接术,吻接处给予人新鲜羊膜包裹,通过大体观察、光镜观察、透射电镜、免疫组化、神经电生理检测、坐骨神经功能指数等指标分析,评价新鲜羊膜促进周围神经损伤后修复再生的效果。方法:选用成年、健康SD大鼠60只,雌雄不限,体重220克~250克。取一侧腿制备坐骨神经急性损伤模型。随机分A组(修复神经后用人新鲜羊膜包裹吻合口),B组(修复神经后用生物膜包裹吻合口),C组(修复神经后不做任何处理)三组,每组20只。术后2、4、8和12周各行大体观察、光镜观察、免疫组织化学检测(s-100蛋白在神经组织中的表达情况);分别于术后4、8、12周各行透射电镜观察、轴突成像分析(计数有髓神经纤维数目、轴突直径及髓鞘厚度)、复合肌肉动作电位检测(测量并记录潜伏期、波幅和神经传导速度)、计算坐骨神经功能指数及其恢复率。所有数据均进行统计学处理。结果:大体观察:三组大鼠术后手术切口都未出现感染,7天后切口拆线,切口一期愈合;三组大鼠术后2周大部出现下肢稍肿胀,活动明显减少且跛行。2周后肿胀、精神状态及饮食逐渐恢复,A、B组大鼠跛行时间较C组短,活动量恢复更早。A、B组和C组相比足底和足趾皮肤溃疡大鼠数量少且溃疡恢复时间短。术后2周羊膜、生物膜局部稍有吸收,4周大部吸收,8周完全吸收。A组、B组神经和周围组织稍有粘连且粘连较疏松,活动度可。C组神经和周围组织广泛紧密粘连,钝性不易分离,活动度差。光镜观察:术后2、4、8、12周A组、B组神经恢复情况明显优于C组。电镜观察:术后4周A、B、C三组神经纤维再生均不明显,板层结构均不清晰。术后8周、12周A、B组比C组神经纤维再生数量更多、髓鞘厚度厚、板层结构更清晰,轴突直径更大。免疫组织化学检测:A、B组中S-100蛋白表达及分布均优于同时期C组。轴突图像分析:A、B组神经吻合口远端有髓神经纤维直径、髓鞘厚度及横截面有髓神经纤维数目均优于同时期C组,结果有统计学意义(P0.05)。神经电生理检测:术后术侧神经的诱发电位潜伏期、波幅及传导速度检测示:A、B组比C组潜伏期短、波幅高及神经传导速度快,结果有统计学意义(P0.05)。坐骨神经功能指数:A组、B组坐骨神经功能指数均明显高于同时期C组,统计学分析有显著性差异(P0.05)。结论:人羊膜作为一种生物材料修复周围神经损伤有比较好的效果。可以减轻受损神经与周围组织的粘连和减少神经吻合处的瘢痕形成;可促进神经纤维再生、轴突直径增大、髓鞘增厚;可以减轻神经切口处炎性反应、免疫反应。
[Abstract]:Objective: peripheral nerve injury is a common disabling disease in the clinic, which directly damages the labor resources and causes a heavy burden on social and family life. The regeneration of peripheral nerve injury is affected by many factors, and the microenvironment and the local scar at the anastomotic stoma seriously affect the repair effect after the nerve injury. For this reason, The improvement of local microenvironment and reduction of local scar is beneficial to the regeneration of peripheral nerve and the recovery of nerve function. The amniotic membrane has semimodialysis, ensures the smooth transmission of nutrients, promotes the proliferation of Schwann cells, accelerates the growth of the nerve axons, inhibits the biological activity of the peripheral fibroblasts, limits the surrounding tissue to invade the nerve broken ends and inhibits the surrounding tissue. In this experiment, the rat sciatic nerve injury model was constructed. The rat model of sciatic nerve injury was constructed, the nerve kissing was carried out under the microscope, and the fresh amniotic membrane was wrapped in the kissing place. The indexes of the fresh sheep were evaluated through gross observation, light microscopy, transmission electron microscopy, immunohistochemistry, neurophysiological test, and the sciatic nerve function index. The effect of membrane on regeneration of peripheral nerve injury. Methods: 60 healthy SD rats, male and female, 220 gram of body weight, 220 gram of body weight were used to prepare the acute injury model of the sciatic nerve in one side of the leg. The group of A (repaired the nerve after the nerve was repaired with the fresh amniotic membrane), and the group B (repair the nerve after the nerve membrane was wrapped with the anastomosis), and the C group (repair) The three groups, 20 rats in each group, were observed in 2,4,8 and 12 weeks after operation, with light microscopy and immunohistochemical staining (the expression of S-100 protein in the nerve tissue); the transmission electron microscopy was performed at the 4,8,12 weeks after the operation, and the axon imaging analysis (counting the number of myelinated nerve fibers, axon diameter and the thickness of myelin sheath) were observed, respectively. Combined muscle action potential detection (measuring and recording latency, amplitude and nerve conduction velocity) and calculating the sciatic nerve function index and its recovery rate. All data were statistically processed. Results: gross observation: three groups of rats were not infected after operation, 7 days after the incision was cut, the incision was healed, and the three groups of rats were 2 weeks after operation. There was a slight swelling of the lower extremities, decreased activity obviously and the limp.2 weeks after the swelling, the mental state and diet gradually resumed. The limping time of the group of A and B rats was shorter than that of the C group, and the amount of activity recovered earlier.A. The number of foot ulcers in the foot and toe of the B group and the C group was less and the recovery time was shorter. The amniotic membrane, the biofilm partially absorbed, and the large part of the biofilm after 2 weeks after the operation. Absorption, 8 weeks completely absorbed group.A, group B nerve and surrounding tissue slightly adhesions and adhesion more loose, activity can be.C group nerve and surrounding tissue widely close adhesion, blunt not easy to separate, poor activity. After 2,4,8,12 week A group, B group nerve recovery is obviously better than the C group. Electron microscope observation: 4 weeks A, B, C three groups of nerve fibers after the operation The regeneration was not obvious, and the lamellar structure was not clear. 8 weeks after the operation, 12 weeks A, the B group had more nerve regeneration than the C group, the thickness of the myelin sheath was thicker, the lamellar structure was clearer and the diameter of the axon was greater. The expression and distribution of S-100 protein in the group of B were better than that of the simultaneous C group. The axonal image analysis: A, B group of distal nerve anastomoses had medullary God. The diameter of the fibers, the thickness of myelin sheath and the number of myelinated nerve fibers in the cross section were better than those in the C group, and the results were statistically significant (P0.05). The neurophysiological tests showed that the latency of evoked potential, amplitude and conduction velocity of the postoperative lateral nerve were detected: A, the latency of the group B was shorter, the amplitude of wave was higher and the nerve conduction velocity was faster, and the results were statistically significant. (P0.05) the sciatic nerve function index: the function index of the sciatic nerve in group A and group B was significantly higher than that of the group C at the same time. There was a significant difference (P0.05). Conclusion: the amniotic membrane as a biological material has a better effect on the repair of peripheral nerve injury. It can reduce the adhesion of the damaged nerve to the surrounding tissue and reduce the nerve anastomosis. Scar formation; can promote nerve fiber regeneration, axon diameter increased, myelin thickening; can reduce the inflammation of nerve incision, immune response.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.3

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