Departmet of Neurosurgery, Xinjiang Autonomous Region People Hospital, Urumqi 830001, China.
Chin Med J (Engl). 2010 Sep;123(17):2424-31.
The most important objective of transplant studies in the injured spinal cord has been to provide a favorable environment for axonal growth. Moreover, the continuing discovery of new grafts is providing new potentially interesting transplant candidates. Our purpose was to observe the morphological and functional repair effects of the co-transplantation of neural stem cell (NSC), Schwann cells (SCs) and poly lactide-co-glycolide acid (PLGA) on the spinal cord injury of rats.
A scaffold of PLGA was fabricated. NSCs and SCs were cultured, with the NSCs labeled with 5-bromodeoxyuridine, and the complex of NSC/PLGA or NSC + SCs/PLGA were constructed. Thirty-six Wistar rats were randomly divided into three groups: group A (transplantation of PLGA), group B (transplantation of NSC/PLGA) and group C (transplantation of NSC + SCs/PLGA). The 3 mm length of the right hemicord was removed under the microscope in all rats. The PLGA or the complex of PLGA-cells were implanted into the injury site. Basso-Beattie-Bresnahan (BBB) locomotion scores, motor and somatosensory evoked potential of lower limbs were examined to learn the rehabilitation of sensory and motor function at 4 weeks, 8 weeks, 12 weeks and 24 weeks after injury. All the recovered spinal cord injury (SCI) tissues were observed with HE staining, immunohistochemistry, and transelectronmicroscopy to identify the survival, migration and differentiation of the transplanted cells and the regeneration of neural fibres at 4 weeks, 8 weeks, 12 weeks and 24 weeks after injury.
(1) From 4 weeks to 24 weeks after injury, the BBB locomotion scores of cell-transplanted groups were better than those of the non-cell-transplanted group, especially group C (P < 0.05). The amplitudes of the somatosensory evoked potential (SEP) and motor-evoked potential (MEP) were improved after injury in groups B and C, but the amplitude of SEP and MEP at 4 weeks was lower than that at 12 weeks and 24 weeks after injury. Compared with group B, the amplitude of SEP and MEP in group C was improved. The amplitude of SEP and MEP was not improved after injury in group A. (2) HE staining revealed the volume of the scaffold decreased and the number of cells in the scaffold increased. Newly-grown capillaries also could be seen. Immunohistochemistry staining showed the transplanted NSCs could survive and migrate until 24 weeks and they could differentiate into neurons and oligodendrocytes. The regenerated axons were observed in the scaffold-cell complex with transelectronmicroscopy. The above manifestations were more extensive in group C.
The transplanted NSC can survive and migrate in the spinal cord of rats up to 24 weeks after injury, and they can differentiate into various neural cells. Co-transplantation of cells/PLGA can promote the functional recovery of the injured spinal cord. The effect of co-transplanting NSC + SCs/PLGA is better than transplanting NSC/PLGA alone.
移植研究的最重要目标是为轴突生长提供有利的环境。此外,不断发现新的移植物为新的潜在有趣的移植候选物提供了可能。我们的目的是观察神经干细胞(NSC)、施万细胞(SCs)和聚乳酸-共-羟基乙酸(PLGA)共移植对大鼠脊髓损伤的形态和功能修复作用。
制备 PLGA 支架。培养 NSCs,用 5-溴脱氧尿苷标记 NSCs,构建 NSC/PLGA 复合物或 NSC+SCs/PLGA 复合物。36 只 Wistar 大鼠随机分为三组:A 组(移植 PLGA)、B 组(移植 NSC/PLGA)和 C 组(移植 NSC+SCs/PLGA)。所有大鼠在显微镜下切除右侧半脊髓 3mm 长。将 PLGA 或 PLGA-细胞复合物植入损伤部位。在损伤后 4 周、8 周、12 周和 24 周,通过 Basso-Beattie-Bresnahan(BBB)运动评分、下肢运动和体感诱发电位检查,了解感觉和运动功能的康复情况。在损伤后 4 周、8 周、12 周和 24 周,用 HE 染色、免疫组织化学和透射电镜观察所有恢复的脊髓损伤(SCI)组织,以鉴定移植细胞的存活、迁移和分化以及神经纤维的再生。
(1)损伤后 4 周至 24 周,细胞移植组的 BBB 运动评分优于非细胞移植组,尤其是 C 组(P<0.05)。B 组和 C 组损伤后体感诱发电位(SEP)和运动诱发电位(MEP)的振幅有所改善,但 4 周时 SEP 和 MEP 的振幅低于 12 周和 24 周时的振幅。与 B 组相比,C 组的 SEP 和 MEP 振幅有所提高。A 组损伤后 SEP 和 MEP 的振幅没有改善。(2)HE 染色显示支架体积减小,支架内细胞数量增加。还可以看到新生长的毛细血管。免疫组织化学染色显示移植的 NSCs 可以存活和迁移,直到损伤后 24 周,它们可以分化为神经元和少突胶质细胞。在支架-细胞复合物中用透射电镜观察到再生轴突。C 组的上述表现更为广泛。
移植的 NSCs 可在损伤后 24 周内在大鼠脊髓内存活和迁移,并可分化为多种神经细胞。细胞/PLGA 的共移植可促进损伤脊髓的功能恢复。共移植 NSC+SCs/PLGA 的效果优于单独移植 NSC/PLGA。