Fluorescence molecular tomography of DiR-labeled mesenchymal stem cell implants for osteochondral defect repair in rabbit knees
Fluorescence molecular tomography of DiR-labeled mesenchymal stem cell implants for osteochondral defect repair in rabbit knees
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DOI：
10.1007/s00330-016-4457-5
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发表时间：
2017-03-01
影响因子：
5.9
通讯作者：
Meier, Reinhard
中科院分区：
文献类型：
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作者：
Berninger, Markus T.;Mohajerani, Pouyan;Meier, Reinhard
为了评估使用近红外染料1，1 '-双十八烷基-3，3，3'标记兔间充质干细胞（MSCs）的效率，3 '-四甲基吲哚三羰花青碘化物（DiR）标记的骨髓间充质干细胞（MSCs）在兔骨软骨缺损修复中的应用（1.25-20 μ g/mL）。通过XTT-和Caspase-3/-7-测试评估细胞凋亡的活力和诱导。通过测量糖胺聚糖评价软骨形成潜力。标记的细胞和未标记的对照（n = 3）在软骨形成分化之前和之后经历FMT-XCT成像。在兔膝关节中通过手术创建骨软骨缺损（n = 6）。将未标记和标记的MSC植入纤维蛋白凝块中并通过FMT-XCT成像。使用多元回归模型进行统计分析，DiR标记的MSC在透射照明模式下的平面图像上产生剂量依赖性荧光信号。在低于10 μ g DiR/mL的浓度下，未检测到存活率或凋亡诱导的显著降低（p > 0.05）; MSC的软骨形成潜力未受到影响（p > 0.05）。标记的MSCs在骨软骨缺损中的FMT-XCT显示了移植的显著信号，（p <0.05），具有关于其骨软骨整合的额外高分辨率解剖信息。FMT-XCT允许检测骨软骨再生过程中的干细胞植入。标记的MSCs显示没有毒副作用或软骨形成的损害。荧光分子断层扫描允许检测MSCs的骨软骨缺损FMT-XCT有助于改善细胞植入和骨软骨再生过程的评估。
To assess labelling efficiency of rabbit mesenchymal stem cells (MSCs) using the near-infrared dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindotricarbocyanine iodide (DiR) and detection of labelled MSCs for osteochondral defect repair in a rabbit model using fluorescence molecular tomography-X-ray computed tomography (FMT-XCT).MSCs were isolated from New Zealand White rabbits and labelled with DiR (1.25-20 mu g/mL). Viability and induction of apoptosis were assessed by XTT- and Caspase-3/-7-testing. Chondrogenic potential was evaluated by measurement of glycosaminoglycans. Labelled cells and unlabeled controls (n = 3) underwent FMT-XCT imaging before and after chondrogenic differentiation. Osteochondral defects were created surgically in rabbit knees (n = 6). Unlabeled and labelled MSCs were implanted in fibrin-clots and imaged by FMT-XCT. Statistical analyses were performed using multiple regression models.DiR-labelling of MSCs resulted in a dose-dependent fluorescence signal on planar images in trans-illumination mode. No significant reduction in viability or induction of apoptosis was detected at concentrations below 10 mu g DiR/mL (p > .05); the chondrogenic potential of MSCs was not affected (p > .05). FMT-XCT of labelled MSCs in osteochondral defects showed a significant signal of the transplant (p < .05) with additional high-resolution anatomical information about its osteochondral integration.FMT-XCT allows for detection of stem cell implantation within osteochondral regeneration processes.DiR-labelling of MSCs shows no toxic side effects or impairment of chondrogenesis.Fluorescence molecular tomography allows for detection of MSCs for osteochondral defect repair.FMT-XCT helps to improve evaluation of cell implantation and osteochondral regeneration processes.