Human amniotic membrane repairs acute sciatic nerve injury in rat models

doi:10.3969/j.issn.2095-4344.1073

Abstract

Abstract:

BACKGROUND: Human amniotic membrane with low immunogenicity can reduce scar formation, inhibit inflammation and prevent angiogenesis.
OBJECTIVE: To observe the effect of human amniotic membrane on repair and regeneration process after acute nerve injury.
METHODS: Ninety adult male Sprague-Dawley rats provided by Laboratory Animal Center of Hebei Medical University were used for establishing the models of acute sciatic nerve injury and then randomly divided into amniotic membrane (nerve entrapment with amniotic membrane), chitosan (nerve entrapment with chitosan gel) and blank control (without any treatment) groups (n=30 per group). Gross, light microscope and transmission electric microscope observations, axon image analysis and electrophysiological examination were performed.
RESULTS AND CONCLUSION: Gross, light microscope and transmission electric microscope observations revealed: compared with the blank control group, the mild limb swelling, neurohyperemia, and edema were observed in the amniotic membrane group. In addition, the adhesion with the surrounding tissue was mild in the amniotic membrane group, the inflammatory reaction was mild and there were few fibroblasts and newborn capillaries. In the amniotic membrane group, function of organelles was active, there were few fibroblasts and less collagen, and the repair time was short, which were similar with the chitosan group. The axon image analysis showed that thickness of the myelin sheath and the diameter of myelinated fibers in the amniotic membrane and chitosan groups were superior to those in the blank control group (P < 0.05). Neurophysiological stimulation showed that there was no significant difference in the nerve conduction velocity among groups. The amplitude was significantly different among groups (P < 0.05). The latent period in the amniotic membrane and chitosan groups was significantly shorter than that in the blank control group (P < 0.05). To conclude, amniotic membrane can provide a relatively stable microenvironment for nerve repair, reduce adhesion, inhibit inflammatory reaction, promote the repair and regeneration of nerve, and show good therapeutic effect. Amniotic membrane has a long retention time in vivo, which is beneficial to nerve regeneration and microenvironment maintenance, and contributes to the recovery of nerve function. Therefore, amniotic membrane is better than the chitosan.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

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

CLC Number:

Cao Peng, Wang Haonan, Tian Weifeng, Sun Naichao, Bai Jiangbo, Yu Kunlun, Tian Dehu. Human amniotic membrane repairs acute sciatic nerve injury in rat models[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(7): 1046-1051.

Figures/Tables 3

2.1 实验动物数量分析 SD大鼠90只，随机分为3组，每组30只，均进入结果分析。 2.2 大体观察结果术后3组大鼠伤口均未见感染，无足底溃疡，未见自噬现像，Ⅰ期愈合。术后1周大鼠左下肢均出现肿胀，拖地现象，肌力差，刺激无收缩，未出现明显缩足及舔足动作。精神状态较差，饮食及尿便较前减少明显。下肢肿胀逐渐减轻，至术后8周羊膜组和几丁糖组跛足现象消失，对外界刺激出现明显缩足及舔足，空白对照组仍有跛足现象，对外界刺激感觉差。术后12周，大鼠对外界刺激敏感，空白对照组稍差。各组大鼠神经周围恢复情况，见图1。术后1，2周，3组大鼠损伤段神经压迹清晰可见，损伤神经菲薄，神经外膜充血，羊膜组和几丁糖组神经与周围组织粘连不明显，空白对照组神经与周围组织有薄膜性的粘连。术后4，8周，神经及周围组织充血水肿减轻，损伤段神经较前增粗，羊膜组及几丁糖组外膜充血基本消退，与周围组织粘连较疏松，空白对照组神经外膜充血较明显，质地较硬，与周围组织粘连较致密。羊膜大部分吸收，几丁糖完全吸收。术后12周，神经及周围组织未见明显充血水肿，损伤段神经增粗，接近正常，神经外膜充血消失，羊膜组和几丁糖组神经与周围组织粘连较疏松，羊膜完全吸收，空白对照组与周围组织粘连广泛而致密，钝性分离困难。 2.3 光镜观察结果见图2。术后1，2周，3组神经排列紊乱，失去正常形态，髓鞘肿胀，可见大量脱髓鞘的神经，视野内出现大量炎性细胞，可见巨噬细胞，其中羊膜组和几丁糖组炎性细胞较少，空白对照组炎性细胞较多。术后4周，羊膜组和几丁糖组神经髓鞘部分恢复正常形态，可见大量许旺细胞。3组神经炎症细胞较前均有所减少，羊膜组和几丁糖组减少更加明显，空白对照组神经细胞大部分未恢复正常形态，炎症细胞仍较多。术后8周，羊膜组和几丁糖组神经排列整齐，炎症细胞明显减少，可见散在许旺细胞核，可见新生毛细血管。空白对照组炎性细胞较多。术后12周，3组神经均恢复正常形态，羊膜组和几丁糖组神经束间较为疏松，空白对照组神经束间较为致密。 2.4 透射电镜观察结果见图3。术后2周，羊膜组和几丁糖组可见髓鞘融合、肿胀、空化，髓鞘不整齐，许旺细胞肿胀，核周系扩张，胞质肿胀，线粒体空泡化，未见明显成纤维细胞。空白对照组可见明显脱髓鞘反应，髓鞘板层结构剥离，髓鞘结构不规则，向内增生，许旺细胞内线粒体空泡化，嵴和膜溶解消失，内质网脱颗粒，核周系扩张，线粒体致密化。偶可见成纤维细胞包裹轴突。术后4周，羊膜组和几丁糖组髓鞘较规则，脱髓鞘反应减少，成纤维细胞及胶原纤维较少，细胞器较空白对照组多。空白对照组髓鞘欠规则，仍可见较多脱髓鞘反应，可见大量成纤维细胞和胶原纤维增生，粗面内质网较多，游离核糖体较发达，线粒体损伤较前减轻。术后8周，羊膜组及几丁糖组成纤维细胞少，轴突排列整齐，粗细较一致，轴突层板较清晰。空白对照组轴突排列较整齐，成纤维细胞和胶原纤维减少。偶可见损伤髓鞘。"

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