Tenomodulin highly expressing MSCs as a better cell source for tendon injury healing.
Tenomodulin highly expressing MSCs as a better cell source for tendon injury healing.
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高表达肌腱调节蛋白的间充质干细胞是肌腱损伤愈合的更好细胞来源。
DOI：
10.18632/oncotarget.20495
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发表时间：
2017-09-29
期刊：
影响因子：
--
通讯作者：
Xu L
中科院分区：
文献类型：
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作者：
Hou Y;Ni M;Lin S;Sun Y;Lin W;Liu Y;Wang H;He W;Li G;Xu L
肌腱损伤是常见的骨科问题，可能会导致严重的发病率。骨髓间充质干细胞（mesenchymal stem cells，MSCs）在组织工程中显示出良好的应用前景，已被用于肌腱损伤的治疗。但骨髓间充质干细胞的成腱分化能力较低，阻碍了其应用。在本研究中，我们构建了Tenomodulin（Tnmd）启动子驱动的GFP表达慢病毒质粒。转染BMSCs后，用流式细胞仪检测GFP的表达，筛选高表达Tnmd的BMSCs。我们发现Tnmd表达水平高的MSCs具有更强的肌腱分化能力。此外，进行RNA测序以鉴定表达较高和较低水平Tnmd的BMSC之间的分子差异。最后，我们证明了GDF7在高表达Tnmd的BMSCs中上调，并在促进BMSCs的成腱分化中起重要作用。内源性GDF7基因沉默可显著抑制骨髓间充质干细胞的肌腱生成，而GDF7基因沉默可显著抑制Tnmd基因表达的骨髓间充质干细胞的肌腱生成，这主要是由于GDF7基因的表达增强了骨髓间充质干细胞的肌腱分化能力。此外，通过大鼠髌腱损伤模型评估具有较高Tnmd水平的BMSCs对肌腱愈合的影响。因此，Tnmd可以作为一种理想的细胞表面标记物，用于筛选具有更高成腱分化能力的BMSCs，而GDF7是MSCs成腱分化所必需的。
Tendon injuries are common orthopedic problems which may cause severe morbidity. MSCs (mesenchymal stem cells) have shown promising effect on tissue engineering and have been used for the treatment of tendon injury. But the low tenogenic differentiation capacity of MSCs have hindered their application. In the present study, we have constructed the Tenomodulin (Tnmd) promoter-driven GFP expression lentiviral plasmid. After transduced into BMSCs, the expression of GFP was used to select BMSCs highly expressing Tnmd by flow cytometry. We found that MSCs with higher level of Tnmd expression had stronger tenogenic differentiation ability. Furthermore, RNA sequencing was performed to identify the molecular difference between BMSCs expressing higher and lower levels of Tnmd. And finally we demonstrated that GDF7 was upregulated in BMSCs highly expressing Tnmd and played an vital role in promoting tenogenic differentiation of BMSCs. GDF7 was mainly accounted for the elevated tenogenic differentiation ability of BMSCs with higher Tnmd expression as silencing the endogenous GDF7 significantly inhibited tenogenesis in BMSCs. In addition, the effect of BMSCs with higher Tnmd level on tendon healing was evaluated by a rat patellar tendon injury model. Taken together, our study showed that Tnmd could be used as an ideal cell surface marker to select cells with higher tenogenic differentiation ability from BMSCs, and GDF7 was indispensable for tenogenesis of MSCs.