Department of Clinical Pharmacology, University Hospital of Tübingen, Tübingen, Germany.
Rejuvenation Res. 2011 Feb;14(1):3-16. doi: 10.1089/rej.2010.1130. Epub 2011 Feb 3.
Safe and effective cell delivery remains one of the main challenges in cell-based therapy of neurodegenerative disorders. Graft survival, sufficient enrichment of therapeutic cells in the brain, and avoidance of their distribution throughout the peripheral organs are greatly influenced by the method of delivery. Here we demonstrate for the first time noninvasive intranasal (IN) delivery of mesenchymal stem cells (MSCs) to the brains of unilaterally 6-hydroxydopamine (6-OHDA)-lesioned rats. IN application (INA) of MSCs resulted in the appearance of cells in the olfactory bulb, cortex, hippocampus, striatum, cerebellum, brainstem, and spinal cord. Out of 1 × 10⁶ MSCs applied intranasally, 24% survived for at least 4.5 months in the brains of 6-OHDA rats as assessed by quantification of enhanced green fluorescent protein (EGFP) DNA. Quantification of proliferating cell nuclear antigen-positive EGFP-MSCs showed that 3% of applied MSCs were proliferative 4.5 months after application. INA of MSCs increased the tyrosine hydroxylase level in the lesioned ipsilateral striatum and substantia nigra, and completely eliminated the 6-OHDA-induced increase in terminal deoxynucleotidyl transferase (TdT)-mediated 2'-deoxyuridine, 5'-triphosphate (dUTP)-biotin nick end labeling (TUNEL) staining of these areas. INA of EGFP-labeled MSCs prevented any decrease in the dopamine level in the lesioned hemisphere, whereas the lesioned side of the control animals revealed significantly lower levels of dopamine 4.5 months after 6-OHDA treatment. Behavioral analyses revealed significant and substantial improvement of motor function of the Parkinsonian forepaw to up to 68% of the normal value 40-110 days after INA of 1 × 10⁶ cells. MSC-INA decreased the concentrations of inflammatory cytokines-interleukin-1β (IL-1β), IL-2, -6, -12, tumor necrosis factor (TNF), interferon-γ (IFN-γ, and granulocyte-macrophage colony-stimulating factor (GM-CSF)-in the lesioned side to their levels in the intact hemisphere. IN administration provides a highly promising noninvasive alternative to the traumatic surgical procedure of transplantation and allows targeted delivery of cells to the brain with the option of chronic application.
安全有效的细胞递送仍然是神经退行性疾病细胞治疗的主要挑战之一。移植物的存活、治疗细胞在大脑中的充分富集以及避免其分布到外周器官，这些都受到递送方法的极大影响。在这里，我们首次证明了非侵入性的鼻腔内（IN）递送至单侧 6-羟多巴胺（6-OHDA）损伤大鼠的脑内的间充质干细胞（MSCs）。MSCs 的 IN 应用（INA）导致细胞出现在嗅球、皮质、海马、纹状体、小脑、脑干和脊髓中。在 IN 应用的 1×10⁶个 MSCs 中，通过增强型绿色荧光蛋白（EGFP）DNA 的定量评估，至少有 24%的细胞在 6-OHDA 大鼠的脑中存活了至少 4.5 个月。对增殖细胞核抗原阳性 EGFP-MSCs 的定量分析表明，应用后 4.5 个月，有 3%的应用 MSCs 具有增殖能力。MSCs 的 INA 增加了损伤同侧纹状体和黑质中的酪氨酸羟化酶水平，并完全消除了 6-OHDA 诱导的这些区域末端脱氧核苷酸转移酶（TdT）介导的 2'-脱氧尿苷、5'-三磷酸（dUTP）-生物素尼克末端标记（TUNEL）染色的增加。EGFP 标记的 MSCs 的 INA 防止了损伤半球中多巴胺水平的任何下降，而对照组动物的损伤侧在 6-OHDA 处理后 4.5 个月显示出明显较低的多巴胺水平。行为分析显示，IN 给予 1×10⁶个细胞后 40-110 天，帕金森病前爪的运动功能显著且实质性改善，达到正常值的 68%。MSC-INA 将损伤侧的炎症细胞因子-白细胞介素-1β（IL-1β）、IL-2、-6、-12、肿瘤坏死因子（TNF）、干扰素-γ（IFN-γ）和粒细胞-巨噬细胞集落刺激因子（GM-CSF）的浓度降低到与未损伤半球相同的水平。IN 给药为创伤性移植手术提供了一种极有前途的非侵入性替代方法，允许将细胞靶向递送至大脑，并可选择进行慢性应用。
