小孔径壳聚糖微载体支持诱导多能干细胞来源神经干细胞的表面黏附生长并改善小鼠嗅觉丧失的鼻内治疗

文献信息

中文摘要

鼻内递送神经干细胞为中枢神经系统修复提供了一种非侵入性策略，但其疗效受到快速黏液纤毛清除和细胞滞留不良的限制。尽管基于壳聚糖的载体已被探索，但指导神经干细胞在鼻内微环境中行为的结构原则仍不明确。我们比较了通过冷冻控制相分离制备的三种孔结构壳聚糖微载体（CSMs）（平均表面孔径分别为4.7±1.7、15.9±3.2和20.5±0.7 µm），并使用诱导多能干细胞来源的神经干细胞（iPSC-NSCs）进行研究。细胞反应并未随孔径线性变化：最小孔径制剂CSM-196支持以表面黏附为主的生长，而两种较大孔径制剂均支持以浸润为主的生长。CSM-196还与增殖活性增加、自发神经元分化以及在没有外源性诱导因子的情况下神经营养因子分泌升高相关。在甲巯咪唑诱导的嗅觉丧失模型中，CSM-196-iPSC-NSC制剂在嗅觉行为恢复、嗅上皮厚度、神经元标志物表达和上皮增殖方面，较以相同途径和方案单独递送细胞产生了更大的改善。这一基于壳聚糖的平台提供了一种以表面黏附为导向的细胞递送策略，值得进一步评估用于鼻内神经再生治疗。

英文摘要

Intranasal delivery of neural stem cells offers a non-invasive strategy for central nervous system repair, yet its efficacy is limited by rapid mucociliary clearance and poor cellular retention. Although chitosan-based carriers have been explored, the architectural principles governing neural stem cell behavior in the intranasal microenvironment remain poorly defined. We compared chitosan microcarriers (CSMs) with three pore architectures (mean surface pore sizes 4.7 ± 1.7, 15.9 ± 3.2, and 20.5 ± 0.7 µm), fabricated via freezing-controlled phase separation, using induced pluripotent stem cell-derived neural stem cells (iPSC-NSCs). Cellular responses did not vary linearly with pore size: the smallest-pore formulation, CSM-196, supported surface-adhesion-dominant growth, whereas both larger-pore formulations supported infiltration-dominant growth. CSM-196 was also associated with increased proliferative activity, spontaneous neuronal differentiation, and elevated neurotrophic factor secretion without exogenous induction factors. In a methimazole-induced anosmia model, the CSM-196-iPSC-NSC formulation produced greater improvements in olfactory behavioral recovery, olfactory epithelial thickness, neuronal marker expression, and epithelial proliferation than cell-only delivery administered by the same route and schedule. This chitosan-based platform provides a surface-adhesion-oriented cell-delivery strategy that warrants further evaluation for intranasal neural regenerative therapy.