Skopalik Josef, Polakova Katerina, Havrdova Marketa, Justan Ivan, Magro Massimiliano, Milde David, Knopfova Lucia, Smarda Jan, Polakova Helena, Gabrielova Eva, Vianello Fabio, Michalek Jaroslav, Zboril Radek
Department of Pharmacology, Masaryk University, Brno, Czech Republic.
Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry and Analytical Chemistry, Faculty of Science, Palacky University, Olomouc, Czech Republic.
Int J Nanomedicine. 2014 Nov 20;9:5355-72. doi: 10.2147/IJN.S66986. eCollection 2014.
Cell therapies have emerged as a promising approach in medicine. The basis of each therapy is the injection of 1-100×10(6) cells with regenerative potential into some part of the body. Mesenchymal stromal cells (MSCs) are the most used cell type in the cell therapy nowadays, but no gold standard for the labeling of the MSCs for magnetic resonance imaging (MRI) is available yet. This work evaluates our newly synthesized uncoated superparamagnetic maghemite nanoparticles (surface-active maghemite nanoparticles - SAMNs) as an MRI contrast intracellular probe usable in a clinical 1.5 T MRI system.
MSCs from rat and human donors were isolated, and then incubated at different concentrations (10-200 μg/mL) of SAMN maghemite nanoparticles for 48 hours. Viability, proliferation, and nanoparticle uptake efficiency were tested (using fluorescence microscopy, xCELLigence analysis, atomic absorption spectroscopy, and advanced microscopy techniques). Migration capacity, cluster of differentiation markers, effect of nanoparticles on long-term viability, contrast properties in MRI, and cocultivation of labeled cells with myocytes were also studied.
SAMNs do not affect MSC viability if the concentration does not exceed 100 μg ferumoxide/mL, and this concentration does not alter their cell phenotype and long-term proliferation profile. After 48 hours of incubation, MSCs labeled with SAMNs show more than double the amount of iron per cell compared to Resovist-labeled cells, which correlates well with the better contrast properties of the SAMN cell sample in T2-weighted MRI. SAMN-labeled MSCs display strong adherence and excellent elasticity in a beating myocyte culture for a minimum of 7 days.
Detailed in vitro tests and phantom tests on ex vivo tissue show that the new SAMNs are efficient MRI contrast agent probes with exclusive intracellular uptake and high biological safety.
细胞疗法已成为医学领域一种很有前景的治疗方法。每种疗法的基础都是将1 - 100×10⁶具有再生潜力的细胞注射到身体的某个部位。间充质基质细胞（MSCs）是目前细胞治疗中最常用的细胞类型，但目前尚无用于磁共振成像（MRI）标记MSCs的金标准。本研究评估了我们新合成的未包覆超顺磁性磁赤铁矿纳米颗粒（表面活性磁赤铁矿纳米颗粒 - SAMNs）作为一种可用于临床1.5 T MRI系统的MRI细胞内造影剂探针。
分离大鼠和人类供体的MSCs，然后在不同浓度（10 - 200 μg/mL）的SAMN磁赤铁矿纳米颗粒中孵育48小时。测试细胞活力、增殖和纳米颗粒摄取效率（使用荧光显微镜、xCELLigence分析、原子吸收光谱和先进的显微镜技术）。还研究了迁移能力、分化标志物簇、纳米颗粒对长期活力的影响、MRI中的造影特性以及标记细胞与心肌细胞的共培养。
如果浓度不超过100 μg铁氧化物/mL，SAMNs不会影响MSCs的活力，且该浓度不会改变其细胞表型和长期增殖特性。孵育48小时后，与用Resovist标记的细胞相比，用SAMNs标记的MSCs每个细胞的铁含量增加了一倍多，这与SAMN细胞样品在T2加权MRI中更好的造影特性密切相关。在搏动的心肌细胞培养中，SAMN标记的MSCs至少7天内表现出强粘附性和出色的弹性。
对离体组织进行的详细体外测试和模型测试表明，新型SAMNs是高效的MRI造影剂探针，具有独特的细胞内摄取和高生物安全性。
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