Wood Matthew D, Willits Rebecca Kuntz
Department of Biomedical Engineering, Saint Louis University, St Louis, MO, USA.
J Neural Eng. 2009 Aug;6(4):046003. doi: 10.1088/1741-2560/6/4/046003. Epub 2009 Jun 3.
Electrical therapies have been found to aid repair of nerve injuries and have been shown to increase and direct neurite outgrowth during stimulation. This enhanced neural growth existed even after the electric field (EF) or stimulation was removed, but the factors that may influence the enhanced growth, such as stimulation media or surface coating, have not been fully investigated. This study characterized neurite outgrowth and branching under various conditions: EF magnitude and application time, ECM surface coating, medium during EF application and growth supplements. A uniform, low-magnitude EF (24 or 44 V m(-1)) was applied to dissociated chick embryo dorsal root ganglia seeded on collagen or laminin-coated surfaces. During the growth period, cells were either exposed to NGF or N2, and during stimulation cells were exposed to either unsupplemented media (Ca(2+)) or PBS (no Ca(2+)). Parallel controls for each experiment included cells exposed to the chamber with no stimulation and cells remaining outside the chamber. After brief electrical stimulation (10 min), neurite length significantly increased 24 h after application for all conditions studied. Of particular interest, increased stimulation time (10-100 min) further enhanced neurite length on laminin but not on collagen surfaces. Neurite branching was not affected by stimulation on any surface, and no preferential growth of neurites was noted after stimulation. Overall, the results of this report suggest that short-duration electric stimulation is sufficient to enhance neurite length under a variety of conditions. While further data are needed to fully elucidate a mechanism for this increased growth, these data suggest that one focus of those investigations should be the interaction between the growth cone and the substrata.
电疗法已被发现有助于神经损伤的修复,并且已证明在刺激过程中能增加并引导神经突生长。即使在去除电场(EF)或刺激后,这种增强的神经生长依然存在,但可能影响增强生长的因素,如刺激介质或表面涂层,尚未得到充分研究。本研究对在各种条件下的神经突生长和分支进行了表征:EF强度和施加时间、细胞外基质(ECM)表面涂层、EF施加期间的培养基以及生长补充剂。将均匀的低强度EF(24或44 V m(-1))施加于接种在胶原蛋白或层粘连蛋白包被表面的解离鸡胚背根神经节。在生长期间,细胞要么暴露于神经生长因子(NGF)要么暴露于N2,在刺激期间细胞要么暴露于未添加补充剂的培养基(Ca(2+))要么暴露于磷酸盐缓冲液(PBS,无Ca(2+))。每个实验的平行对照包括未接受刺激的置于培养箱中的细胞以及留在培养箱外的细胞。经过短暂电刺激(10分钟)后,在所研究的所有条件下,施加刺激24小时后神经突长度显著增加。特别值得注意的是,增加刺激时间(10 - 100分钟)进一步增强了层粘连蛋白表面而非胶原蛋白表面的神经突长度。神经突分支在任何表面上均不受刺激影响,刺激后也未观察到神经突的优先生长。总体而言,本报告结果表明,短时间电刺激足以在多种条件下增强神经突长度。虽然需要更多数据来充分阐明这种生长增加的机制,但这些数据表明这些研究的一个重点应该是生长锥与基质之间的相互作用。