Nanoscaled and microscaled parallel topography promotes tenogenic differentiation of ASC and neotendon formation in vitro.
Nanoscaled and microscaled parallel topography promotes tenogenic differentiation of ASC and neotendon formation in vitro.
复制标题
纳米级和微米级平行形貌促进体外 ASC 的腱分化和新腱形成
DOI：
10.2147/ijn.s161423
复制
发表时间：
2018
影响因子：
8
通讯作者：
Liu W
中科院分区：
文献类型：
--
作者：
Zhou K;Feng B;Wang W;Jiang Y;Zhang W;Zhou G;Jiang T;Cao Y;Liu W
背景不同尺度的地形在指导间充质干细胞包括脂肪干细胞（adipose-derived stem cells，ASCs）的分化中起着重要作用，其分化效应仍有待进一步研究。目的探讨微尺度和纳米尺度排列形貌诱导人成体干细胞（hASCs）成腱分化的异同。方法以平行微槽PDMS膜和平行排列的明胶/聚己内酯混合物电纺纳米纤维为模型进行研究。结果细胞形态学定量分析（细胞面积、胞体方向和胞体主轴角）显示，微米级和纳米级的排列形貌均能诱导细胞平行排列，形成细长的细胞形态。qPCR分析还表明，在两种尺度下对齐的拓扑结构可以在体外培养的第7天诱导各种肌腱形成标志物的基因表达，包括腱调节蛋白、胶原I和胶原VI、核心蛋白聚糖、腱生蛋白-C和双糖蛋白聚糖，但仅在微尺度拓扑结构中上调了巩膜轴和腱生蛋白-C的表达。此外，在第3天的tenogenic分化仅在微尺度上得到证实。此外，微尺度拓扑结构被确认为在组织水平上的肌腱诱导，因为在与小鼠ASC体外培养后，形成了具有成熟的I型胶原纤维的证据的新肌腱组织，所述I型胶原纤维仅在平行排列的聚乙醇酸（PGA）微纤维中。相反，在随机图案化的PGA微纤维中仅形成脂肪组织。结论微尺度和纳米尺度排列的形貌均能诱导hASCs向腱样细胞分化，微尺度形貌较纳米尺度形貌更能诱导细胞形态伸长和稳定的腱样标志物表达。微尺度的诱导作用也证实了在组织水平上的新腱形成在体外。
Background Topography at different scales plays an important role in directing mesenchymal stem cell differentiation including adipose-derived stem cells (ASCs) and the differential effect remains to be investigated. Purpose This study aimed to investigate the similarity and difference between micro- and nanoscaled aligned topography for inducing tenogenic differentiation of human ASCs (hASCs). Methods Parallel microgrooved PDMS membrane and a parallel aligned electrospun nanofibers of gelatin/poly-ε-caprolactone mixture were employed as the models for the study. Results Aligned topographies of both microscales and nanoscales could induce an elongated cell shape with parallel alignment, as supported by quantitative cell morphology analysis (cell area, cell body aspect, and cell body major axis angle). qPCR analysis also demonstrated that the aligned topography at both scales could induce the gene expressions of various tenogenic markers at the 7th day of in vitro culture including tenomodulin, collagen I and collagen VI, decorin, tenascin-C and biglycan, but with upregulated expression of scleraxis and tenascin-C only in microscaled topography. Additionally, tenogenic differentiation at the 3rd day was confirmed only at microscale. Furthermore, microscaled topography was confirmed for its tenogenic induction at tissue level as neotendon tissue was formed with the evidence of mature type I collagen fibers only in parallel aligned polyglycolic acid (PGA) microfibers after in vitro culture with mouse ASCs. Instead, only fat tissue was formed in random patterned PGA microfibers. Conclusion Both microscaled and nanoscaled aligned topographies could induce tenogenic differentiation of hASCs and micro-scaled topography seemed better able to induce elongated cell shape and stable tenogenic marker expression when compared to nanoscaled topography. The microscaled inductive effect was also confirmed at tissue level by neotendon formation in vitro.