High Osteogenic Potential of Adipose- and Muscle-derived Mesenchymal Stem Cells in Spinal-Ossification Model Mice.
High Osteogenic Potential of Adipose- and Muscle-derived Mesenchymal Stem Cells in Spinal-Ossification Model Mice.
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DOI：
10.1097/brs.0000000000002266
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发表时间：
2017-12-01
期刊：
影响因子：
3
通讯作者：
Ishibashi Y
中科院分区：
文献类型：
--
作者：
Liu X;Kumagai G;Wada K;Tanaka T;Asari T;Oishi K;Fujita T;Mizukami H;Furukawa KI;Ishibashi Y
小鼠后纵韧带骨化(OPLL)模型的基础实验。目的评价从肌肉和脂肪组织中获取的间充质干细胞(MSCs)在脚尖行走(TTW)小鼠的成骨能力，在TTW小鼠中，颈椎OPLL压迫脊髓并导致运动和感觉障碍。在人类中，间充质干细胞参与了宫颈OPLL的发病机制。TTW小鼠的颈椎OPLL会导致脊髓的慢性压迫。很少有研究将骨髓间充质干细胞成骨潜能与OPLL动物模型中的行为变化进行比较。我们比较了TTW小鼠(4-20周龄)和对照野生型(无骨质增生症)小鼠的MSCs的成骨能力和行为特征。术后8周用计算机断层扫描和病理检查观察韧带骨化情况，并用抗骨髓间充质干细胞标志物(CD45和CD105)的荧光抗体对组织进行双重染色。使用Basso Mouse量表评估运动功能，并使用热和机械测试评估感觉功能。通过茜素红S吸光度、成骨矿化染色和实时定量聚合酶链式反应检测成骨相关基因，评价脂肪和肌肉间充质干细胞的成骨能力。TTW小鼠在8周龄时开始脊柱韧带骨化，骨化面积随年龄增长而增大。免疫荧光染色显示骨化区有MSCs。TTW小鼠在8周龄时变得迟钝，Basso老鼠量表评分显示12周龄时开始出现运动功能缺陷。茜素红S矿化染色显示，在4、8和20周龄时，TTW小鼠的脂肪和肌肉来源的MSCs的成骨潜能高于野生型小鼠。实时定量聚合酶链式反应显示TTW MSCs强表达成骨相关基因。TTW小鼠肌肉和脂肪组织来源的MSCs具有较高的成骨潜能。证据级别：不适用
Basic experiments in a mouse model of ossification of the posterior longitudinal ligament (OPLL). To assess the osteogenic potential of mesenchymal stem cells (MSCs) obtained from muscle and adipose tissue in Tiptoe-walking (ttw) mice, in which cervical OPLL compresses the spinal cord and causes motor and sensory dysfunction. In humans, MSCs have been implicated in the pathogenesis of cervical OPLL. Cervical OPLL in ttw mice causes chronic compression of the spinal cord. Few studies have compared the MSC osteogenic potential with behavioral changes in an OPLL animal model. We compared the osteogenic potential and behavioral characteristics of MSCs from ttw mice (4 to 20 weeks old) with those from control wild-type mice (without hyperostosis). Ligament ossification was monitored by micro-computed tomography and pathology; tissues were double stained with fluorescent antibodies against markers for MSCs (CD45 and CD105), at 8 weeks. The Basso Mouse Scale was used to assess motor function, and heat and mechanical tests to assess sensory function. The osteogenic potential of adipose and muscle MSCs was assessed by Alizarin Red S absorbance, staining for osteogenic mineralization, and real-time quantitative polymerase chain reaction for osteogenesis-related genes. Spinal-ligament ossification began in ttw mice at 8 weeks of age, and the ossified area increased with age. Immunofluorescence staining identified MSCs in the ossification area. The ttw mice became hyposensitive at 8 weeks of age, and Basso Mouse Scale scores showed motor-function deficits starting at 12 weeks of age. Alizarin Red S staining for mineralization showed a higher osteogenic potential in the adipose- and muscle-derived MSCs from ttw mice than from wild-type mice at 4, 8, and 20 weeks of age. Real-time quantitative polymerase chain reaction showed that ttw MSCs strongly expressed osteogenesis-related genes. MSCs derived from muscle and adipose tissue in ttw mice had a high osteogenic potential. Level of Evidence: N/A