Zheng Yijing, Zhou Yifei, Zhang Xiaolei, Chen Yuemiao, Zheng Xuhao, Cheng Tao, Wang Chaonan, Hu Xuqi, Hong Jianjun
Department of Orthopedics, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, P.R. China.
Department of Hematology, Wenzhou Central Hospital, Wenzhou, Zhejiang 325000, P.R. China.
Exp Ther Med. 2017 Jun;13(6):3195-3202. doi: 10.3892/etm.2017.4383. Epub 2017 Apr 26.
Achilles tendons have a very poor capacity for intrinsic regeneration. The cell-based treatment strategy for Achilles tendinitis includes the application of mesenchymal stem cells (MSCs), which have high proliferative and multipotent differentiation ability, and is a promising approach. The aim of the present study was to explore the tenogenic potential of human menstrual blood stromal stem cells (MenSCs) in a co-culture system and to compare the tenogenic capability under normoxic and hypoxic conditions. MenSCs were co-cultured indirectly with Achilles tendon cells in a Transwell co-culture system for 1, 2, or 3 weeks in two different concentrations of oxygen (20 and 2% O), whereas the control contained only MenSCs. The extracellular matrix of MenSCs in each system was evaluated by Alcian blue staining assay, histological staining, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and western blot analysis. Alcian blue staining assay revealed a significant increase (P<0.05) in proteoglycan secretion by the differentiated MenSCs. Identical results were obtained by RT-qPCR for collagen I, which was validated by western blot analysis. Considerably increased collagen I and collagen III gene expression levels were exhibited by cells in the co-culture treatment group when compared with the control (P<0.05); however, no significant difference was detected between the normoxic (20% O) and hypoxic treatment (2% O) groups. RT-qPCR was utilized to determine the expression levels of thrombospondin 4, scleraxis and tenascin C in the differentiated MenSCs; a significant increase in the expression of these specific genes was indicated in the co-culture treatment group compared with the control (P<0.05). Although the expression levels were markedly higher in hypoxia than in normoxia conditions, this difference was not significant. To conclude, the present study indicated that MenSCs manifested a strong proliferative and multipotent capacity for differentiation and differentiated into Achilles tenogenic cells. Therefore, the use of MenSCs may be considered in Achilles tendinitis therapy.
跟腱的自身再生能力非常差。基于细胞的跟腱炎治疗策略包括应用间充质干细胞（MSCs），其具有高增殖和多能分化能力，是一种很有前景的方法。本研究的目的是在共培养系统中探索人月经血基质干细胞（MenSCs）的成腱潜力，并比较常氧和低氧条件下的成腱能力。在Transwell共培养系统中，将MenSCs与跟腱细胞间接共培养1、2或3周，设置两种不同的氧浓度（20%O₂和2%O₂），而对照组仅含MenSCs。通过阿尔辛蓝染色试验、组织学染色、逆转录-定量聚合酶链反应（RT-qPCR）和蛋白质印迹分析对各系统中MenSCs的细胞外基质进行评估。阿尔辛蓝染色试验显示，分化后的MenSCs蛋白聚糖分泌显著增加（P<0.05）。RT-qPCR检测I型胶原得到相同结果，蛋白质印迹分析验证了该结果。与对照组相比，共培养治疗组细胞的I型胶原和III型胶原基因表达水平显著升高（P<0.05）；然而，常氧（20%O₂）和低氧治疗（2%O₂）组之间未检测到显著差异。利用RT-qPCR测定分化后MenSCs中血小板反应蛋白4、硬骨素和腱生蛋白C的表达水平；与对照组相比，共培养治疗组这些特定基因的表达显著增加（P<0.05）。虽然低氧条件下的表达水平明显高于常氧条件，但这种差异并不显著。总之，本研究表明MenSCs表现出强大的增殖和多能分化能力，并分化为跟腱成腱细胞。因此，在跟腱炎治疗中可考虑使用MenSCs。
