Department of Anesthesiology, Emory University School of Medicine, Atlanta, GA, USA.
Cell Transplant. 2013;22(11):2001-15. doi: 10.3727/096368912X657909. Epub 2012 Oct 12.
Transplantation of bone marrow mesenchymal stem cells (BMSCs) provides a promising regenerative medicine for stroke. Whether BMSC therapy could repair ischemia-damaged neuronal circuits and recover electrophysiological activity has largely been unknown. To address this issue, BMSCs were implanted into the ischemic barrel cortex of adult mice 1 and 7 days after focal barrel cortex stroke. Two days after the first transplantation (3 days after stroke), the infarct volume determined by TTC staining was significantly smaller in BMSC-treated compared to vehicle-treated stroke mice. The behavioral corner test showed better long-term recovery of sensorimotor function in BMSC-treated mice. Six weeks poststroke, thalamocortical slices were prepared and neuronal circuit activity in the peri-infarct region of the barrel cortex was determined by extracellular recordings of evoked field potentials. In BMSC-transplanted brain slices, the ischemia-disrupted intracortical activity from layer 4 to layer 2/3 was noticeably recovered, and the thalamocortical circuit connection was also partially restored. In contrast, much less and slower recovery was seen in control animals of barrel cortex stroke. Immunohistochemical staining disclosed that the density of neurons, axons, and blood vessels in the peri-infarct region was significantly higher in BMSC-treated mice, accompanied with enhanced local blood flow recovery. Western blotting showed that BMSC treatment increased the expression of stromal cell-derived factor-1 (SDF-1), vascular endothelial growth factor (VEGF), and brain-derived neurotrophic factor (BDNF) in the peri-infarct region. Moreover, the expression of the axonal growth associated protein-43 (GAP-43) was markedly increased, whereas the axonal growth inhibiting proteins ROCK II and NG2 were suppressed in the BMSC-treated brains. BMSC transplantation also promoted directional migration and survival of doublecortin (DCX)-positive neuroblasts in the peri-infarct region. The present investigation thus provides novel evidence that BMSC transplantation has the potential to repair the ischemia-damaged neural networks and restore lost neuronal connections. The recovered circuit activity likely contributes to the improved sensorimotor function after focal ischemic stroke and BMSC transplantation.
骨髓间充质干细胞(BMSCs)移植为中风提供了一种有前途的再生医学方法。BMSC 疗法是否能够修复缺血性损伤的神经元回路并恢复电生理活性,在很大程度上尚不清楚。为了解决这个问题,在局灶性大脑皮层卒中后 1 天和 7 天,将 BMSCs 移植到成年小鼠的缺血性大脑皮层桶状皮层中。第一次移植后两天(卒中后 3 天),与 vehicle 处理的卒中小鼠相比,TTC 染色确定的梗死体积在 BMSC 处理的小鼠中明显较小。角测试显示,BMSC 处理的小鼠在感觉运动功能方面有更好的长期恢复。卒中后 6 周,制备丘脑皮质切片,并通过皮层桶状区梗死周围的诱发电场电位的细胞外记录来确定神经元回路活动。在 BMSC 移植的脑片中,从第 4 层到第 2/3 层的缺血性破坏的皮层内活动明显恢复,并且丘脑皮质回路连接也部分恢复。相比之下,在对照动物的大脑皮层卒中中,恢复的程度要低得多,速度也要慢得多。免疫组织化学染色显示,在 BMSC 处理的小鼠中,梗死周围区的神经元、轴突和血管密度明显更高,并且局部血流恢复增强。Western blot 显示,BMSC 处理增加了基质细胞衍生因子-1(SDF-1)、血管内皮生长因子(VEGF)和脑源性神经营养因子(BDNF)在梗死周围区的表达。此外,轴突生长相关蛋白-43(GAP-43)的表达明显增加,而在 BMSC 处理的大脑中,轴突生长抑制蛋白 ROCK II 和 NG2 的表达受到抑制。BMSC 移植还促进了双皮质素(DCX)阳性神经前体细胞在梗死周围区的定向迁移和存活。本研究因此提供了新的证据,表明 BMSC 移植具有修复缺血性损伤的神经网络和恢复丢失的神经元连接的潜力。恢复的回路活动可能有助于改善局灶性缺血性卒中和 BMSC 移植后的感觉运动功能。