Sukhinich Kirill K, Dashinimaev Erdem B, Vorotelyak Ekaterina A, Aleksandrova Maria A
Koltzov Institute of Developmental Biology, Russian Academy of Sciences, Moscow, Russia.
Pirogov Russian National Research Medical University, Moscow, Russia.
Plast Reconstr Surg Glob Open. 2020 Feb 11;8(2):e2610. doi: 10.1097/GOX.0000000000002610. eCollection 2020 Feb.
The regeneration of the peripheral nerves after injuries is still a challenging fundamental and clinical problem. The cell therapy and nerve guide conduit construction are promising modern approaches. Nowadays, different sources of cells for transplantation are available. But it is little known about the interaction between fetal central nervous system cells and peripheral nerve tissue. In this study, we analyzed the development of the fetal neocortex and spinal cord solid grafts injected into the gelatin hydrogel conduits and their effects on sciatic nerve regeneration after cut injury.
Frontal neocortex tissue was obtained from E19.5 and spinal cord tissue was obtained from E14.5 fetuses harvested from transgenic EGFP mice. The grafts were injected into the hydrogel conduits which were connected to the nerve stumps after cut injury. The recovery of motor function was estimated with walking track analysis at 2, 5, and 8 weeks after surgery. Then immunohistochemical study was performed.
The histological examination showed that only fetal neocortex solid graft cells had survived after implantation. Immunostaining revealed that some of the transplanted cells expressed neural markers such as neurofilament protein and NeuN. But the cells mostly differentiated in glial lineage, which was confirmed with immunostaining for GFAP and S100β. The walking-track analysis has shown that 8 weeks after surgery bioengineered conduit differed significantly from the control.
We revealed that the hydrogel conduit is suitable for nerve re-growth and that the fetal neocortex grafted cells can survive and differentiate. Bioengineered conduit can stimulate functional recovery after the nerve injury.
周围神经损伤后的再生仍然是一个具有挑战性的基础和临床问题。细胞治疗和神经导管构建是很有前景的现代方法。如今，有不同来源的细胞可用于移植。但关于胎儿中枢神经系统细胞与周围神经组织之间的相互作用却知之甚少。在本研究中，我们分析了注入明胶水凝胶导管的胎儿新皮质和脊髓实体移植物的发育情况及其对坐骨神经切断损伤后再生的影响。
从E19.5的胎儿获取额叶新皮质组织，从转基因EGFP小鼠的E14.5胎儿获取脊髓组织。将移植物注入水凝胶导管，导管在切断损伤后与神经残端相连。在术后2周、5周和8周通过行走轨迹分析评估运动功能的恢复情况。然后进行免疫组织化学研究。
组织学检查显示，植入后仅胎儿新皮质实体移植物细胞存活。免疫染色显示，一些移植细胞表达神经标志物，如神经丝蛋白和神经元核抗原（NeuN）。但这些细胞大多分化为胶质细胞谱系，这通过胶质纤维酸性蛋白（GFAP）和S100β的免疫染色得到证实。行走轨迹分析表明，术后8周生物工程导管与对照组有显著差异。
我们发现水凝胶导管适合神经再生，胎儿新皮质移植细胞能够存活并分化。生物工程导管可刺激神经损伤后的功能恢复。
