中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (18): 2890-2899.doi: 10.3969/j.issn.2095-4344.2017.18.017

• 细胞外基质材料 extracellular matrix materials • 上一篇    下一篇

新鲜羊膜修复急性坐骨神经损伤

马  韬,寇文冠,刘国立,白江博,于昆仑,田德虎   

  1. 河北医科大学第三医院,河北省石家庄市  050000
  • 收稿日期:2017-01-16 出版日期:2017-06-28 发布日期:2017-07-07
  • 通讯作者: 田德虎,博士,主任医师,教授,河北医科大学第三医院,河北省石家庄市 050000
  • 作者简介:马韬,男,1989年生,河北省张家口市人,汉族,硕士,主要从事手外科学方面的研究。
  • 基金资助:

    河北省自然科学基金(ZL20140066)

Fresh amniotic membrane for repair of acute peripheral nerve injury

Ma Tao, Kou Wen-guan, Liu Guo-li, Bai Jiang-bo, Yu Kun-lun, Tian De-hu
  

  1. Third Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China
  • Received:2017-01-16 Online:2017-06-28 Published:2017-07-07
  • Contact: Tian De-hu, M.D., Chief physician, Professor, Third Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China
  • About author:Ma Tao, Master, Third Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China
  • Supported by:

    the Natural Science Foundation of Hebei Province, No. ZL20140066

摘要:

文章快速阅读:

 

文题释义:
瓦勒溃变
:神经轴突断裂将会致使损伤平面以远的轴突得不到胞体的营养物质,因此损伤平面以远的神经纤维将最终全部发生崩解,同时大量许旺细胞分裂增殖并且重新形成新的细胞索(Bungner氏带),Bungner氏带可以引导再生轴突延伸并最终进入终末效应器组织,此过程被称为瓦勒溃变。瓦勒溃变形成的微环境十分适宜轴突再生。
许旺细胞:是周围神经系统的胶质细胞,它包绕周围神经轴突,参与并维持神经周围内环境的稳态并且在周围神经损伤、修复与再生中起着至关重要的作用。主要作用有:①激活单核/巨噬细胞吞噬轴突和髓鞘的残片,形成Bungner带支持、引导新生的轴突按正确的方向由近侧段向远端延伸;②分泌多种神经营养因子在神经损伤、修复、再生过程中起到维持受损神经元活性和显著促进神经轴突生长等作用;③许旺细胞膜表面表达多种黏附因子促进轴突再生;④随着神经损伤的修复,增殖的许旺细胞会将轴突完全包裹最终形成成熟的髓鞘。

背景: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。
目的:评价新鲜羊膜促进周围神经损伤后再生的效果。
方法:选用成年SD大鼠60只,制备单侧坐骨神经急性损伤模型,随机分为人羊膜组、生物膜组、空白对照组,每组20只,均在显微镜下行神经吻接术,吻接处给予人新鲜羊膜、生物膜包裹,空白对照组修复神经后不做任何处理。术后2,4,8,12周行大体观察、光镜观察、免疫组织化学检测;术后4,8,12周行透射电镜观察、轴突成像分析、复合肌肉动作电位检测、坐骨神经功能指数测定。
结果与结论:①大体观察: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。人羊膜组、生物膜组神经和周围组织稍有粘连且粘连较疏松,活动度可。空白对照组神经和周围组织广泛紧密粘连,钝性不易分离,活动度差;②光镜观察:术后2,4,8,12周,人羊膜组、生物膜组神经恢复情况明显优于空白对照组;③电镜观察:术后4周,3组神经纤维再生均不明显,板层结构均不清晰。术后8,12周,与空白对照组比较,人羊膜组、生物膜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;④免疫组织化学检测:人羊膜组、生物膜组中S-100蛋白表达及分布均优于同时期空白对照组;⑤轴突图像分析:人羊膜组、生物膜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空白对照组,差异有显著性意义(P < 0.05);⑥神经电生理检测:与空白对照组比较,人羊膜组、生物膜组潜伏期短、波幅高及神经传导速度快,差异有显著性意义(P < 0.05);⑦坐骨神经功能指数:人羊膜组、生物膜组坐骨神经功能指数均明显高于同时期空白对照组,差异有显著性意义(P < 0.05);⑧结果表明,人羊膜作为一种生物材料修复周围神经损伤效果较好,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。

关键词: 生物材料, 材料相容性, 人羊膜, 生物膜, 周围神经损伤, 神经电生理, 免疫组织化学, 河北省自然科学基金

Abstract:

BACKGROUND: To improve local microenvironment and reduce local scars is conducive to peripheral nerve regeneration that promotes nerve function recovery.
OBJECTIVE: To evaluate the effect of fresh amniotic membrane on the regeneration of tinjured peripheral nerve.
METHODS: Sixty healthy Sprague-Dawley rats were randomly divided into three groups (n=20 per group) after constructing a model of sciatic nerve injury of the unilateral leg. In group A, the nerve was wrapped with fresh human amnion at the anastomosis end after the repair of nerve. In group B, the nerve was wrapped with biofilm at the anastomosis end after the repair of nerve. In group C, no treatment was conducted after the repair of nerve (blank control). The effects were evaluated by anatomical observation, light microscope observation, immunohistochemical detection (2, 4, 8, 12 weeks after surgery), transmission electron microscope observation, axon imaging analysis, action potential detection, and sciatic nerve function index (4, 8, 12 weeks after surgery).
RESULTS AND CONCLUSION: (1) Gross observation. The amniotic membrane and biofilm were absorbed partially at postoperative 2 weeks, mostly at postoperative 4 weeks and completely at postoperative 8 weeks. In the groups A and B, the nerve was adhered slightly and loosely to the surrounding tissues, with a fair range of motion. In the group C, the nerve was tightly adhered to the surrounding tissues, with a poor range of motion. (2) Observation under light microscope. The nerve regeneration was better in the groups A and B than group C at 2, 4, 8, 12 weeks postoperatively. (3) Observation under electron microscope. Regenerated nerve fibers were rarely seen and lamellar structures were unclear in the three groups at 4 weeks postoperatively. Then, increased regenerated nerve fibers, thickened myelin sheath, clear lamellar structure and enlarged axon diameter were found in the groups A and B compared with the group C at 8 and 12 weeks postoperatively. (4) Immunohistochemical detection. The expression and distribution of S-100 protein in the groups A and B were better than those in the group C. (5) Axon image analysis. Groups A and B were superior to the group C in the diameter of myelinated nerve fibers, thickness of myelin sheath and number of regenerated nerve fibers. There was a significant difference by statistical analysis (P < 0.05). (6) Electrophysiological examination. Shorter latency period, higher amplitude and faster nerve conduction velocities were observed in the groups A and B compared with the group C (P < 0.05). (7) The sciatic function index. The sciatic function index in group A or B was significantly higher than that in group C (P < 0.05). To conclude, the human amniotic membrane can reduce adhesion between the damaged nerve and surrounding tissues, and prevent scarring at the anastomosis end. In addition, it promotes the regeneration of nerve fibers, increase axon diameter and myelin sheath thickness, and ease inflammatory and immune responses at the neural incision.

Key words: Amnion, Biofilms, Sciatic Nerve, Nerve Regeneration, Tissue Engineering

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