Shi R, Borgens R B, Blight A R
Center for Paralysis Research, Department of Basic Medical Sciences, Purdue University, West Lafayette, Indiana, USA.
J Neurotrauma. 1999 Aug;16(8):727-38. doi: 10.1089/neu.1999.16.727.
We describe a technique using the water-soluble polymer polyethylene glycol (PEG) to reconnect the two segments of completely transected mammalian spinal axons within minutes. This was accomplished by fusing completely severed strips of isolated guinea pig thoracic white matter maintained in vitro in a double sucrose gap recording chamber. The faces of the severed segments were pressed together, and PEG (MW 1,400-3,500 d; approximately 50% by weight in distilled water) was applied directly to this region through a micropipette and removed by aspiration within 2 min. Successful fusion was documented by the immediate restored conduction of compound action potentials through the original transection and by the variable numbers of fused axons in which anatomical continuity was shown to be restored by high-resolution light microscopy and by the diffusion of intracellular fluorescent dyes through fused axons. These data support the conclusion that some severed and subsequently PEG-fused spinal axons both demonstrate restored anatomical continuity and also are physiologically competent to conduct action potentials. This work adds to our previous demonstration that PEG application can immediately repair severely crushed, rather than cut, spinal cord white matter, and may lead to novel treatments for acute trauma to the central and peripheral nervous systems.
我们描述了一种技术,该技术使用水溶性聚合物聚乙二醇(PEG)在数分钟内重新连接完全横断的哺乳动物脊髓轴突的两个部分。这是通过在双蔗糖间隙记录室中融合体外培养的分离豚鼠胸段白质的完全切断的条带实现的。将切断部分的表面压在一起,通过微量移液器将PEG(分子量1400 - 3500道尔顿;在蒸馏水中约50%重量)直接应用于该区域,并在2分钟内通过抽吸去除。通过复合动作电位立即恢复通过原始横断处的传导,以及通过高分辨率光学显微镜显示解剖连续性恢复的融合轴突数量可变,以及细胞内荧光染料通过融合轴突扩散,记录了成功的融合。这些数据支持以下结论:一些切断后经PEG融合的脊髓轴突既显示出恢复的解剖连续性,在生理上也能够传导动作电位。这项工作补充了我们之前的证明,即应用PEG可以立即修复严重挤压而非切断的脊髓白质,并且可能为中枢和周围神经系统的急性创伤带来新的治疗方法。