Tracking of autologous adipose tissue-derived mesenchymal stromal cells with in vivo magnetic resonance imaging and histology after intralesional treatment of artificial equine tendon lesions--a pilot study.
Tracking of autologous adipose tissue-derived mesenchymal stromal cells with in vivo magnetic resonance imaging and histology after intralesional treatment of artificial equine tendon lesions--a pilot study.
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DOI：
10.1186/s13287-016-0281-8
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发表时间：
2016-02-01
影响因子：
7.5
通讯作者：
Stadler PM
中科院分区：
文献类型：
--
作者：
Geburek F;Mundle K;Conrad S;Hellige M;Walliser U;van Schie HT;van Weeren R;Skutella T;Stadler PM
脂肪组织来源的间充质基质细胞 (AT-MSC) 经常用于治疗马肌腱病。到目前为止，关于自体 AT-MSCs 病灶内注射到马浅屈肌腱 (SDFT) 后的命运的知识非常有限。本研究的目的是通过站立磁共振成像 (MRI) 和组织学，在 3 至 9 周的交错时间内监测病灶内注射的用超顺磁性氧化铁 (SPIO) 纳米粒子和绿色荧光蛋白 (GFP) 标记的自体 AT-MSC 的存在。四匹成年温血马接受单侧注射 10 × 106 个自体 AT-MSC 到手术创建的前肢 SDFT 病变中。在三匹马中施用的 AT-MSC 表达慢病毒转导的 GFP 报告基因，并与 SPIO 颗粒共同标记。在治疗后 3、5、7 和 9 周，通过对两匹马进行站立低场 MRI 重复检查以及对所有马进行尸检，使用普鲁士蓝染色、荧光显微镜检查以及使用抗 GFP 抗体进行免疫荧光和免疫组织化学评估 SDFT 中 AT-MSC 的存在。在每次 MRI 中，在 T2* 和 T1 加权序列中，直到观察期结束，在处理过的 SDFT 中都可以检测到标记有 SPIO 颗粒的 AT-MSC。尸检显示，所有处理过的肌腱均含有大量 SPIO 和 GFP 标记的细胞。站立低场 MRI 有可能成功追踪 SPIO 标记的 AT-MSC。组织学、荧光显微镜、免疫荧光和免疫组织化学是在病灶内注射到手术创建的马 SDFT 病灶后检测标记 AT-MSC 的有效工具。病灶内注射 10 × 106 AT-MSC 会导致手术造成的肌腱病灶内及其周围存在大量 AT-MSC，持续时间长达 9 周。将注射的 AT-MSC 整合到愈合的肌腱组织中是病灶内给药后的重要途径。必须谨慎选择注射技术以避免注射的细胞底物回流。体内低场 MRI 可用作非侵入性工具，用于监测患有 SDFT 肌腱病的马体内 AT-MSC 的归巢和植入。
Adipose tissue-derived mesenchymal stromal cells (AT-MSCs) are frequently used to treat equine tendinopathies. Up to now, knowledge about the fate of autologous AT-MSCs after intralesional injection into equine superficial digital flexor tendons (SDFTs) is very limited. The purpose of this study was to monitor the presence of intralesionally injected autologous AT-MSCs labelled with superparamagnetic iron oxide (SPIO) nanoparticles and green fluorescent protein (GFP) over a staggered period of 3 to 9 weeks with standing magnetic resonance imaging (MRI) and histology. Four adult warmblood horses received a unilateral injection of 10 × 106 autologous AT-MSCs into surgically created front-limb SDFT lesions. Administered AT-MSCs expressed lentivirally transduced reporter genes for GFP and were co-labelled with SPIO particles in three horses. The presence of AT-MSCs in SDFTs was evaluated by repeated examinations with standing low-field MRI in two horses and post-mortem in all horses with Prussian blue staining, fluorescence microscopy and with immunofluorescence and immunohistochemistry using anti-GFP antibodies at 3, 5, 7 and 9 weeks after treatment. AT-MSCs labelled with SPIO particles were detectable in treated SDFTs during each MRI in T2*- and T1-weighted sequences until the end of the observation period. Post-mortem examinations revealed that all treated tendons contained high numbers of SPIO- and GFP-labelled cells. Standing low-field MRI has the potential to track SPIO-labelled AT-MSCs successfully. Histology, fluorescence microscopy, immunofluorescence and immunohistochemistry are efficient tools to detect labelled AT-MSCs after intralesional injection into surgically created equine SDFT lesions. Intralesional injection of 10 × 106 AT-MSCs leads to the presence of high numbers of AT-MSCs in and around surgically created tendon lesions for up to 9 weeks. Integration of injected AT-MSCs into healing tendon tissue is an essential pathway after intralesional administration. Injection techniques have to be chosen deliberately to avoid reflux of the cell substrate injected. In vivo low-field MRI may be used as a non-invasive tool to monitor homing and engraftment of AT-MSCs in horses with tendinopathy of the SDFT.