Rodrigues J M, Luís A L, Lobato J V, Pinto M V, Lopes M A, Freitas M, Geuna S, Santos J D, Maurício A C
Centro de Estudos de Ciência Animal (CECA), Instituto de Ciências e Tecnologias Agrárias e Agro-Alimentares (ICETA) da Universidade do Porto, Campus Agrário de Vairão, Rua Padre Armando Quintas, 4485-661 Vairão, Portugal.
Biomed Mater Eng. 2005;15(6):455-65.
Entubulation repair of peripheral nerve injuries has a lengthy history. Several experimental and clinical studies have explored the effectiveness of many biodegradable and non-degradable tubes with or without addition of molecules and cells. The main objective of the present study was to develop an economical and also an easy way for culturing a neural cell line which was capable of growing, differentiating and producing locally nerve growth factors that are otherwise extremely expensive, inside 90 PLA/10 PLG nerve guides. For this purpose the authors have chosen the N1E-115 cell line, a clone of cells derived from mouse neuroblastoma C-1300 with the perspective of using this differentiated cellular system to cover the inside of 90 PLA/10 PLG nerve guides placed to bridge a gap in the rat sciatic nerve experimental model. The N1E-115 cells proliferate in normal culture medium but undergo neuronal differentiation in response to DMSO. Upon induction of differentiation, proliferation of N1E-115 cells ceases, extensive neurite outgrowth is observed and the membranes become highly excitable. While it is known that Ca2+ serves as an important intracellular signal for various cellular processes, such as growth and differentiation. It is also known that can be toxic to cells and is involved in the triggering of events leading to excitotoxic cell death in neurons. The [Ca2+]i in non-differentiated N1E-115 cells and after distinct periods of differentiation, have been determined by the epifluorescence technique using the Fura-2-AM probe. The results of this quantitative assessment revealed that N1E-115 cells which undergo neuronal differentiation for 48 hours in the presence of 1.5% DMSO are best qualified to be used to cover the interior of the nerve guides since the [Ca2+]i was not found to be elevated indicating thus that the onset the cell death processes was not occurred.
周围神经损伤的插管修复已有很长的历史。多项实验和临床研究探讨了许多可生物降解和不可生物降解的导管在添加或不添加分子及细胞情况下的有效性。本研究的主要目的是开发一种经济且简便的方法，用于在90%聚乳酸/10%聚乙醇酸神经导管内培养一种神经细胞系，该细胞系能够生长、分化并产生局部神经生长因子，而这些因子原本极其昂贵。为此，作者选择了N1E - 115细胞系，它是从小鼠神经母细胞瘤C - 1300衍生而来的细胞克隆，目的是利用这个分化的细胞系统覆盖置于大鼠坐骨神经实验模型间隙处的90%聚乳酸/10%聚乙醇酸神经导管内部。N1E - 115细胞在正常培养基中增殖，但对二甲基亚砜（DMSO）有反应时会发生神经元分化。诱导分化后，N1E - 115细胞的增殖停止，观察到广泛的神经突生长，且细胞膜变得高度兴奋。虽然已知钙离子（Ca2+）作为各种细胞过程（如生长和分化）的重要细胞内信号。但也已知其对细胞可能有毒性，并参与引发导致神经元兴奋性毒性细胞死亡的事件。使用Fura - 2 - AM探针通过落射荧光技术测定了未分化的N1E - 115细胞以及分化不同时间段后的细胞内钙离子浓度（[Ca2+]i）。这种定量评估的结果表明，在1.5% DMSO存在下经历48小时神经元分化的N1E - 115细胞最适合用于覆盖神经导管内部，因为未发现细胞内钙离子浓度升高，这表明细胞死亡过程尚未开始。
