Department of Biomedical Engineering, University of Michigan , Ann Arbor, MI 48109-2099, USA.
Tissue Eng Part C Methods. 2009 Dec;15(4):541-50. doi: 10.1089/ten.tec.2008.0302.
The purpose of this study was to evaluate the effects of alginate composition on the neurotrophic factor release, viability, and proliferation of encapsulated neural stem cells (NSCs), as well as on the mechanical stability of the scaffold itself. Four compositions were tested: a high guluronic acid (68%) and a high mannuronic acid (54%) content alginate, with or without a poly-L-lysine (PLL) coating layer. Enzyme-linked immunosorbent assay was used to quantify the release of brain-derived neurotrophic factor, glial-derived neurotrophic factor, and nerve growth factor from the encapsulated cells. All three factors were detected from encapsulated cells only when a high L-guluronic acid alginate without PLL was used. Additionally, capsules with this composition remained intact more frequently when exposed to solutions of low osmolarity, potentially indicating superior mechanical stability. Alginate beads with a PLL-coated, high D-mannuronic acid composition were the most prone to breakage in the osmotic pressure test, and were too fragile for histology and proliferation assays after 1 week in vitro. NSCs survived and proliferated in the three remaining alginate compositions similarly over the 21-day study course irrespective of scaffold condition. NSC-seeded alginate beads with a high L-guluronic acid, non-PLL-coated composition may be useful in the repair of injured nervous tissue, where the mechanism is the secretion of neuroprotective factors. We verify the neuroprotective effects of medium conditioned by NSC-seeded alginate beads on the serum withdrawal-mediated death of PC-12 cells here.
本研究旨在评估藻酸盐组成对神经干细胞(NSCs)包封后神经营养因子释放、活力和增殖的影响,以及对支架本身机械稳定性的影响。测试了四种组成:高古罗糖醛酸(68%)和高甘露糖醛酸(54%)含量的藻酸盐,有无聚-L-赖氨酸(PLL)涂层。酶联免疫吸附试验用于定量检测包封细胞释放脑源性神经营养因子、胶质源性神经营养因子和神经生长因子。只有使用不含 PLL 的高 L-古罗糖醛酸藻酸盐时,才能从包封细胞中检测到所有三种因子。此外,当暴露在低渗透压溶液中时,具有这种组成的胶囊更频繁地保持完整,这可能表明机械稳定性更高。具有 PLL 涂层的高 D-甘露糖醛酸组成的藻酸盐珠在渗透压试验中最容易破裂,并且在体外 1 周后,其组织学和增殖测定结果表明其过于脆弱。在 21 天的研究过程中,无论支架条件如何,NSCs 在其余三种藻酸盐组成中均能存活并增殖。具有高 L-古罗糖醛酸、非 PLL 涂层组成的 NSC 包封藻酸盐珠可能有助于受损神经组织的修复,其机制是分泌神经保护因子。在这里,我们验证了 NSC 包封藻酸盐珠条件培养基对 PC-12 细胞血清撤离介导的死亡的神经保护作用。