Department of Laboratory Medicine, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing Drum Tower Hospital, Nanjing, Jiangsu 210008, P.R. China.
Department of Medical Imaging, Shanghai Jiahui International Hospital, Shanghai 200233, P.R. China.
Mol Med Rep. 2020 Oct;22(4):3327-3337. doi: 10.3892/mmr.2020.11431. Epub 2020 Aug 13.
Endothelial progenitor cells (EPCs) have been discovered to be relevant to the prognosis of cardiovascular diseases. Previous research has demonstrated that EPCs serve vital roles in the occurrence and development of atherosclerosis. Significant improvements have been made in MRI technology and in the experimental use of EPCs for therapeutic angiogenesis and vascular repair. Nevertheless, the migratory, adhesive, proliferative and angiogenic properties of EPCs remain unknown. The aims of the present study were to investigate the potential of using non‑invasive monitoring with ultrasmall superparamagnetic iron oxide nanoparticle (USPION)‑labeled endothelial progenitor cells (EPCs) after transplantation, and to assess the treatment outcomes in an atherosclerotic rabbit model. EPCs derived from rabbit peripheral blood samples were labeled with USPION‑poly‑l‑lysine (USPION‑PLL). The morphology, proliferation, adhesive ability and labeling efficiency of the EPCs were determined by optical and electron microscopy. Moreover, biological activity was assessed by flow cytometry. In addition, T2‑weighted image fast spin‑echo MRI was used to detect cell labeling. USPION content in the labeled EPCs was determined by Prussian blue staining and scanning electron microscopy. Rabbit atherosclerosis model was established using a high‑fat diet. USPION‑labeled EPCs were transplanted into rabbits, and in vivo MRI was performed 1 and 7 days after transplantation. It was found that EPCs cultured on Matrigel formed capillary‑like structures, and expressed the surface markers CD133, CD31, CD34 and vascular endothelial growth factor receptor 2 (VEGFR2). The optimal USPION concentration was 32 µg/ml, as determined by adhesion and proliferation assays. It was identified that USPION‑PLL nanoparticles were 10‑20 nm in diameter. Histopathological analysis results indicated that 1 day after transplantation of the labeled EPCs, blue‑stained granules were observed in the intima of vascular lesions in rabbit models after Prussian blue staining. Therefore, the present results suggest that USPION‑labeled EPCs may play a role in repairing endothelial injury and preventing atherosclerosis in vivo.
内皮祖细胞 (EPCs) 已被发现与心血管疾病的预后相关。先前的研究表明,EPCs 在动脉粥样硬化的发生和发展中起着至关重要的作用。MRI 技术和 EPC 治疗性血管生成和血管修复的实验应用取得了重大进展。然而,EPC 的迁移、黏附、增殖和血管生成特性仍不清楚。本研究旨在探讨使用超顺磁性氧化铁纳米颗粒(USPION)标记的内皮祖细胞(EPCs)移植后进行非侵入性监测的潜力,并评估其在动脉粥样硬化兔模型中的治疗效果。从兔外周血样本中分离出 EPCs,并用 USPION-聚-L-赖氨酸(USPION-PLL)进行标记。通过光学显微镜和电子显微镜观察 EPCs 的形态、增殖、黏附能力和标记效率。此外,通过流式细胞术评估其生物学活性。此外,还使用 T2 加权快速自旋回波 MRI 检测细胞标记。通过普鲁士蓝染色和扫描电子显微镜测定标记 EPCs 中的 USPION 含量。使用高脂饮食建立兔动脉粥样硬化模型。将 USPION 标记的 EPCs 移植到兔体内,移植后 1 天和 7 天进行体内 MRI 检查。结果发现,EPCs 在 Matrigel 上培养形成毛细血管样结构,并表达表面标志物 CD133、CD31、CD34 和血管内皮生长因子受体 2(VEGFR2)。通过黏附和增殖试验确定最佳 USPION 浓度为 32μg/ml。结果表明,USPION-PLL 纳米颗粒的直径为 10-20nm。组织病理学分析结果表明,移植标记的 EPCs 1 天后,普鲁士蓝染色兔模型血管病变内膜可见蓝色颗粒。因此,本研究结果表明,USPION 标记的 EPCs 可能在体内修复内皮损伤和预防动脉粥样硬化中发挥作用。