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使用葡聚糖包被的超顺磁性氧化铁纳米颗粒对牙髓干细胞进行体外和体内MRI追踪

MRI-Tracking of Dental Pulp Stem Cells In Vitro and In Vivo Using Dextran-Coated Superparamagnetic Iron Oxide Nanoparticles.

作者信息

Zare Shahrokh, Mehrabani Davood, Jalli Reza, Saeedi Moghadam Mahdi, Manafi Navid, Mehrabani Golshid, Jamhiri Iman, Ahadian Samad

机构信息

Stem Cell Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Fars 71348-14336, Iran.

Department of Biochemistry, School of Biotechnology and Agriculture, Shiraz Branch, Islamic Azad University, Shiraz, Fars 71987-74731, Iran.

出版信息

J Clin Med. 2019 Sep 9;8(9):1418. doi: 10.3390/jcm8091418.

Abstract

The aim of this study was to track dental pulp stem cells (DPSCs) labeled with dextran-coated superparamagnetic iron oxide nanoparticles (SPIONs) using magnetic resonance imaging (MRI). Dental pulp was isolated from male Sprague Dawley rats and cultured in Dulbecco's modified Eagle's medium F12 (DMEM-F12) and 10% fetal bovine serum. Effects of SPIONs on morphology, viability, apoptosis, stemness, and osteogenic and adipogenic differentiation of DPSCs were assessed. Prussian blue staining and MRI were conducted to determine in vitro efficiency of SPIONs uptake by the cells. Both non-labeled and labeled DPSCs were adherent to culture plates and showed spindle-shape morphologies, respectively. They were positive for osteogenic and adipogenic induction and expression of cluster of differentiation (CD) 73 and CD90 biomarkers, but negative for expression of CD34 and CD45 biomarkers. The SPIONs were non-toxic and did not induce apoptosis in doses less than 25 mg/mL. Internalization of the SPIONs within the DPSCs was confirmed by Prussian blue staining and MRI. Our findings revealed that the MRI-based method could successfully monitor DPSCs labeled with dextran-coated SPIONs without any significant effect on osteogenic and adipogenic differentiation, viability, and stemness of DPSCs. We provided the in vitro evidence supporting the feasibility of an MRI-based method to monitor DPSCs labeled with SPIONs without any significant reduction in viability, proliferation, and differentiation properties of labeled cells, showing that internalization of SPIONs within DPSCs were not toxic at doses less than 25 mg/mL. In general, the SPION labeling does not seem to impair cell survival or differentiation. SPIONs are biocompatible, easily available, and cost effective, opening a new avenue in stem cell labeling in regenerative medicine.

摘要

本研究的目的是使用磁共振成像(MRI)追踪用葡聚糖包被的超顺磁性氧化铁纳米颗粒(SPIONs)标记的牙髓干细胞(DPSCs)。从雄性Sprague Dawley大鼠中分离牙髓,并在杜氏改良 Eagle 培养基 F12(DMEM-F12)和 10%胎牛血清中培养。评估了 SPIONs 对 DPSCs 的形态、活力、凋亡、干性以及成骨和成脂分化的影响。进行普鲁士蓝染色和 MRI 以确定细胞摄取 SPIONs 的体外效率。未标记和标记的 DPSCs 均分别贴附于培养板并呈现纺锤形形态。它们对成骨和成脂诱导以及分化簇(CD)73 和 CD90 生物标志物的表达呈阳性,但对 CD34 和 CD45 生物标志物的表达呈阴性。在剂量小于 25 mg/mL 时,SPIONs 无毒且不诱导凋亡。普鲁士蓝染色和 MRI 证实了 SPIONs 在 DPSCs 内的内化。我们的研究结果表明,基于 MRI 的方法可以成功监测用葡聚糖包被的 SPIONs 标记的 DPSCs,而对 DPSCs 的成骨和成脂分化、活力和干性没有任何显著影响。我们提供了体外证据,支持基于 MRI 的方法监测用 SPIONs 标记的 DPSCs 的可行性,且标记细胞的活力、增殖和分化特性没有任何显著降低,表明在剂量小于 25 mg/mL 时,SPIONs 在 DPSCs 内的内化无毒。总体而言,SPION 标记似乎不会损害细胞存活或分化。SPIONs 具有生物相容性、易于获得且成本效益高,为再生医学中的干细胞标记开辟了一条新途径。

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