Department of Orthopedic Surgery, Hirosaki University Graduate School of Medicine, Hirosaki, Aomori, Japan.
Department of Pathology and Molecular Medicine, Hirosaki University Graduate School of Medicine, Hirosaki, Aomori, Japan.
Spine (Phila Pa 1976). 2017 Dec 1;42(23):E1342-E1349. doi: 10.1097/BRS.0000000000002266.
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.
N/A.
在后纵韧带骨化（OPLL）鼠模型中的基础实验。
评估提踵行走（ttw）小鼠肌肉和脂肪组织来源的间充质干细胞（MSCs）的成骨潜能，在 ttw 小鼠中，颈椎 OPLL 压迫脊髓并导致运动和感觉功能障碍。
在人类中，MSCs 被认为与颈椎 OPLL 的发病机制有关。ttw 小鼠的颈椎 OPLL 导致脊髓慢性受压。在 OPLL 动物模型中，很少有研究比较 MSC 成骨潜能与行为变化。
我们比较了 ttw 小鼠（4 至 20 周龄）和对照野生型小鼠（无骨肥厚）的 MSCs 的成骨潜能和行为特征。通过微计算机断层扫描和组织病理学监测韧带骨化；在 8 周时，用荧光抗体双重染色 MSCS 的标志物（CD45 和 CD105）。使用 Basso 小鼠量表评估运动功能，使用热和机械测试评估感觉功能。通过茜素红 S 吸收、成骨矿化染色和实时定量聚合酶链反应评估脂肪和肌肉来源 MSC 的成骨潜能。
脊柱韧带骨化始于 ttw 小鼠 8 周龄，骨化面积随年龄增加而增加。免疫荧光染色鉴定出骨化区的 MSCs。ttw 小鼠在 8 周龄时变得低敏感，Basso 小鼠量表评分显示运动功能障碍始于 12 周龄。4、8 和 20 周龄时，ttw 小鼠来源的脂肪和肌肉 MSC 的茜素红 S 染色显示出更高的成骨潜能。实时定量聚合酶链反应显示 ttw MSC 强烈表达成骨相关基因。
ttw 小鼠肌肉和脂肪组织来源的 MSCs 具有较高的成骨潜能。
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