Ramalho Bruna Dos Santos, Almeida Fernanda Martins de, Sales Conrado Mendonça, de Lima Silmara, Martinez Ana Maria Blanco
Laboratório de Neurodegeneração e Reparo, Departamento de Patologia - Faculdade de Medicina, HUCFF, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Neural Regen Res. 2018 Jun;13(6):1046-1053. doi: 10.4103/1673-5374.233448.
In spite of advances in surgical care and rehabilitation, the consequences of spinal cord injury (SCI) are still challenging. Several experimental therapeutic strategies have been studied in the SCI field, and recent advances have led to the development of therapies that may act on the inhibitory microenvironment. Assorted lineages of stem cells are considered a good treatment for SCI. This study investigated the effect of systemic transplantation of mesenchymal stem cells (MSCs) in a compressive SCI model. Here we present results of the intraperitoneal route, which has not been used previously for MSC administration after compressive SCI. We used adult female C57BL/6 mice that underwent laminectomy at the T level, followed by spinal cord compression for 1 minute with a 30-g vascular clip. The animals were divided into five groups: sham (anesthesia and laminectomy but without compression injury induction), MSC i.p. (intraperitoneal injection of 8 × 10 MSCs in 500 µL of DMEM at 7 days after SCI), MSC i.v. (intravenous injection of 8 × 10 MSCs in 500 µL of DMEM at 7 days after SCI), DMEM i.p. (intraperitoneal injection of 500 µL of DMEM at 7 days after SCI), DMEM i.v. (intravenous injection of 500 µL of DMEM at 7 days after SCI). The effects of MSCs transplantation in white matter sparing were analyzed by luxol fast blue staining. The number of preserved fibers was counted in semithin sections stained with toluidine blue and the presence of trophic factors was analyzed by immunohistochemistry. In addition, we analyzed the locomotor performance with Basso Mouse Scale and Global Mobility Test. Our results showed white matter preservation and a larger number of preserved fibers in the MSC groups than in the DMEM groups. Furthermore, the MSC groups had higher levels of trophic factors (brain-derived neurotrophic factor, nerve growth factor, neurotrophin-3 and neurotrophin-4) in the spinal cord and improved locomotor performance. Our results indicate that injection of MSCs by either intraperitoneal or intravenous routes results in beneficial outcomes and can be elected as a choice for SCI treatment.
尽管外科治疗和康复取得了进展，但脊髓损伤（SCI）的后果仍然具有挑战性。在SCI领域已经研究了几种实验性治疗策略，最近的进展导致了可能作用于抑制性微环境的疗法的发展。各种干细胞谱系被认为是治疗SCI的良好方法。本研究调查了间充质干细胞（MSCs）全身移植在压迫性SCI模型中的效果。在这里，我们展示了腹腔内途径的结果，该途径以前未用于压迫性SCI后MSCs的给药。我们使用成年雌性C57BL/6小鼠，在T水平进行椎板切除术，然后用30克血管夹对脊髓进行1分钟的压迫。将动物分为五组：假手术组（麻醉和椎板切除术，但未诱导压迫性损伤）、MSCs腹腔注射组（SCI后7天腹腔注射500μL DMEM中8×10个MSCs）、MSCs静脉注射组（SCI后7天静脉注射500μL DMEM中8×10个MSCs）、DMEM腹腔注射组（SCI后7天腹腔注射500μL DMEM）、DMEM静脉注射组（SCI后7天静脉注射500μL DMEM）。通过Luxol固蓝染色分析MSCs移植对白质保留的影响。在甲苯胺蓝染色的半薄切片中计数保留纤维的数量，并通过免疫组织化学分析营养因子的存在。此外，我们用Basso小鼠量表和整体运动测试分析运动性能。我们的结果显示，与DMEM组相比，MSCs组白质保留且保留纤维数量更多。此外，MSCs组脊髓中营养因子（脑源性神经营养因子、神经生长因子、神经营养素-3和神经营养素-4）水平更高，运动性能得到改善。我们的结果表明，通过腹腔内或静脉内途径注射MSCs均可产生有益结果，可作为SCI治疗的一种选择。
