Lu Siliang, Li Ke, Yang Yinxiang, Wang Qian, Yu Yu, Wang Zhaoyan, Luan Zuo
The First Clinical Medical College, Guangxi Medical University, Nanning, 530021, Guangxi, People's Republic of China.
Laboratory of Pediatrics, The Sixth Medical Center of PLA General Hospital, Beijing, 100048, People's Republic of China.
Neuropsychiatr Dis Treat. 2022 Feb 23;18:413-426. doi: 10.2147/NDT.S350586. eCollection 2022.
Stem cell administration via the intranasal route has shown promise as a new therapy for hypoxic-ischemic encephalopathy (HIE). In this study, we aimed to improve the intranasal delivery of stem cells to the brain.
Human neural stem cells (hNSCs) were identified using immunofluorescence, morphological, and flow cytometry assays before transplantation, and cell migration capacity was examined using the transwell assay. Cerebral hypoxia-ischemia (HI) was induced in 7-day-old rats, followed by the intranasal transplantation of CM-Dil-labeled hNSCs. We examined various experimental conditions, including preconditioning hNSCs with hypoxia, catheter method, multiple low-dose transplantation, head position, cell appropriate concentration, and volume. Rats were sacrificed 1 or 3 days after the final intranasal administration, and parts of the nasal tissue and whole brain sections were analyzed under a fluorescence microscope.
The isolated hNSCs met the characteristics of neural stem cells. Hypoxia (5% O, 24 h) enhanced the surface expression of CXC chemokine receptor 4 (CXCR4) (9.21 ± 1.9% ~ 24.76 ± 2.24%, P < 0.01) on hNSCs and improved migration (toward stromal cell-derived factor 1 [SDF-1], 0.54 ± 0.11% ~ 8.65 ± 1.76%, P < 0.001; toward fetal bovine serum, 8.36 ± 0.81% ~ 21.74 ± 0.85%, P < 0.0001). Further improvement increased the number of surviving cell distribution with increased uniformity on the olfactory epithelium and allowed the cells to stay in the nasal cavity for at least 72 h, but they did not survive for longer than 48 h. Optimization of pre-transplantation conditions augmented the success rate of intranasally delivered cells to the brain (0-41.6%). We also tentatively identified that hNSCs crossed the olfactory epithelium into the tissue space below the lamina propria, with cerebrospinal fluid entering the cribriform plate into the subarachnoid space, and then migrated toward injured areas along the brain blood vessels.
This study offers some helpful advice and reference for addressing the problem of repeatability in the intranasal delivery of stem cells.
经鼻途径给予干细胞已显示出作为缺氧缺血性脑病（HIE）新疗法的潜力。在本研究中，我们旨在改善干细胞经鼻向脑内的递送。
在移植前，通过免疫荧光、形态学和流式细胞术检测鉴定人神经干细胞（hNSCs），并使用transwell检测法检测细胞迁移能力。在7日龄大鼠中诱导脑缺氧缺血（HI），随后经鼻移植CM-Dil标记的hNSCs。我们研究了各种实验条件，包括用缺氧预处理hNSCs、导管法、多次低剂量移植、头部位置、细胞合适浓度和体积。在最后一次经鼻给药后1天或3天处死大鼠，在荧光显微镜下分析部分鼻组织和全脑切片。
分离的hNSCs符合神经干细胞的特征。缺氧（5% O₂，24小时）增强了hNSCs表面CXC趋化因子受体4（CXCR4）的表达（9.21±1.9%24.76±2.24%，P<0.01），并改善了迁移（向基质细胞衍生因子1[SDF-1]迁移，0.54±0.11%8.65±1.76%，P<0.001；向胎牛血清迁移，8.36±0.81%~21.74±0.85%，P<0.0001）。进一步的改进增加了存活细胞分布的数量，提高了嗅上皮的均匀性，并使细胞在鼻腔中停留至少72小时，但它们存活不超过48小时。优化移植前条件提高了经鼻递送细胞进入脑内的成功率（0-41.6%）。我们还初步确定hNSCs穿过嗅上皮进入固有层下方的组织间隙，脑脊液进入筛板进入蛛网膜下腔，然后沿脑血管向损伤区域迁移。
本研究为解决干细胞经鼻递送的可重复性问题提供了一些有益的建议和参考。
