Department of Functional and Systems Neurobiology, Cajal Institute, CSIC Madrid, Spain.
J Biomater Appl. 2012 Mar;26(7):791-809. doi: 10.1177/0885328210379928. Epub 2010 Sep 28.
Chitosan (Ch) and some of its derivatives have been proposed as good biomaterials for tissue engineering, to construct scaffolds promoting tissue regeneration. In this work we made composite films from Ch and mixtures of Ch with gelatin (G) and poly-l-lysine (PLL), and evaluated the growth on these films of PC12 and C6 lines as well as neurons and glial cells derived from cerebral tissue and dorsal root ganglia (DRG). C6 glioma cells proliferated on Ch, G, and Ch + G films, although metabolic activity was decreased by the presence of the G in the mixtures. NGF-differentiated PC12 cells, adhered preferentially on Ch and films containing PLL. Unlike NGF-treated PC12 cells, cortical and hippocampal neurons showed good adhesion to Ch and Ch + G films, where they extended neurites. Astrocytes adhered on Ch, Ch + G, and Ch + PLL mixtures, although viability decreased during the culture time. Olfactory ensheathing cells (OEC) adhered and proliferated to confluency on the wells covered with Ch + G films. Neurites from DRGs exhibited high extension on these films. These results demonstrate that Ch + G films have excellent adhesive properties for both neurons and regeneration-promoting glia (OEC). These films also promoted neurite extension from DRG, making them good candidates for tissue engineering of nerve repair.
壳聚糖(Ch)及其一些衍生物已被提议作为组织工程的良好生物材料,用于构建促进组织再生的支架。在这项工作中,我们制备了壳聚糖与明胶(G)和聚-l-赖氨酸(PLL)混合物的复合膜,并评估了 PC12 和 C6 细胞系以及源自脑组织和背根神经节(DRG)的神经元和神经胶质细胞在这些膜上的生长情况。C6 神经胶质瘤细胞在壳聚糖、G 和 Ch+G 膜上增殖,尽管混合物中存在 G 会降低代谢活性。与 NGF 处理的 PC12 细胞不同,神经生长因子分化的 PC12 细胞优先附着在壳聚糖和含有 PLL 的膜上。与 NGF 处理的 PC12 细胞不同,皮质和海马神经元在壳聚糖和 Ch+G 膜上具有良好的粘附性,在这些膜上它们可以延伸神经突。星形胶质细胞附着在壳聚糖、Ch+G 和 Ch+PLL 混合物上,尽管在培养过程中细胞活力下降。嗅鞘细胞(OEC)附着在覆盖有 Ch+G 膜的孔上并增殖至汇合。来自 DRG 的神经突在这些膜上表现出高度的延伸。这些结果表明,Ch+G 膜对神经元和促进再生的神经胶质(OEC)均具有极好的粘附性能。这些膜还促进了来自 DRG 的神经突延伸,使其成为神经修复组织工程的良好候选材料。