In vitro biocompatibility of chitosan-based materials to primary culture of hippocampal neurons
In vitro biocompatibility of chitosan-based materials to primary culture of hippocampal neurons
复制标题
壳聚糖基材料对海马神经元原代培养物的体外生物相容性
DOI：
10.1007/s10856-009-3702-8
复制
发表时间：
2009-07-01
影响因子：
3.7
通讯作者：
Ding, Fei
中科院分区：
文献类型：
--
作者：
He, Qianru;Zhang, Tianyi;Ding, Fei
天然生物材料壳聚糖是一种很有前途的神经引导导管材料，广泛应用于周围神经修复。本研究旨在探讨壳聚糖对原代培养的海马神经元的生物相容性。由壳聚糖纤维或膜制成的基底被发现支持所附海马神经元的存活和生长，通过使用光学和电子显微镜以及神经丝200、生长相关蛋白-43、微管相关蛋白2、β-微管蛋白III和突触素的免疫细胞化学。MTT法显示，培养不同时间后，壳聚糖纤维或膜提取液中的海马神经元细胞活力与羟基磷灰石提取液或普通神经元培养液中的海马神经元细胞活力无显著性差异，但明显高于有机锡提取液。Western分析显示，在壳聚糖提取物和普通神经元培养基中培养的海马神经元之间，生长相关蛋白-43和beta-tubulin III的蛋白水平没有显着差异。这些结果共同表明，壳聚糖是生物相容性的原代培养的海马神经元细胞表型和功能没有细胞毒性作用，提高了潜在的可能性，使用壳聚糖中枢神经系统治疗。
The natural biomaterial chitosan has been widely used as a promising nerve guidance conduit material for peripheral nerve repair. This study aimed to investigate in vitro biocompatibility of chitosan to primarily cultured hippocampal neurons, one type of central nervous system (CNS) cells. The substrate made up of chitosan fibers or membranes was found to support the survival and growth of the attached hippocampal neurons by using light and electron microscopy as well as immunocytochemistry for neurofilament 200, growth-associated protein-43, microtubule-associated protein 2, beta-tubulin III and synaptophysin. MTT assay indicated that the cell viability of hippocampal neurons in chitosan fiber or membrane extract was not significantly different from that in hydroxyapatite extract or plain neuronal medium, but significantly higher than that in organotin extract after culture for different times. Western analysis revealed that no significant difference in the protein level of growth-associated protein-43 and beta-tubulin III was detected between hippocampal neurons cultured in chitosan extract and in plain neuronal culture medium. The results collectively demonstrate that chitosan is biocompatible to primary culture of hippocampal neurons without cytotoxic effects on cell phenotype and functions, raising a potential possibility of using chitosan for CNS therapy.