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首页> 外文期刊>Acta biomaterialia >Hybrid-spheroids incorporating ECM like engineered fragmented fibers potentiate stem cell function by improved cell/cell and cell/ECM interactions
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Hybrid-spheroids incorporating ECM like engineered fragmented fibers potentiate stem cell function by improved cell/cell and cell/ECM interactions

机译：掺入ECM的混合球状体，如工程化的碎片纤维增强干细胞功能，通过改善的细胞/细胞和细胞/ ECM相互作用

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Graphical abstract Display Omitted Abstract Extracellular matrix (ECM) microenvironment is critical for the viability, stemness, and differentiation of stem cells. In this study, we developed hybrid-spheroids of human turbinate mesenchymal stem cells (hTMSCs) by using extracellular matrix (ECM) mimicking fragmented fibers (FFs) for improvement of the viability and functions of hTMSCs. We prepared FFs with average size of 68.26 μm by partial aminolysis of poly L-lactide (PLLA) fibrous sheet (FS), which was coated with polydopamine for improved cell adhesion. The proliferation of hTMSCs within the hybrid-spheroids mixed with fragmented fibers was significantly increased as compared to that from the cell-only group. Cells and fragmented fibers were homogenously distributed with the presence of pore like empty spaces in the structure. LOX-1 staining revealed that the hybrid-spheroids improved the cell viability, which was potentially due to enhanced transport of oxygen through void space generated by engineered ECM. Transmission electron microscopy (TEM) analysis confirmed that cells within the hybrid-spheroid formed strong cell junctions and contacts with fragmented fibers. The expression of cell junction proteins including connexin 43 and E-cadherin was significantly upregulated in hybrid-spheroids by 16.53?±?0.04 and 28.26?±?0.11-fold greater than that from cell-only group. Similarly, expression of integrin α 2, α 5 , and β 1 was significantly enhanced at the same group by 25.72?±?0.13, 27.48?±?0.49, and 592.78?±?0.06-fold, respectively. In addition, stemness markers including Oct-4, Nanog, and Sox2 were significantly upregulated in hybrid-spheroids by 96.56?±?0.06, 158.95?±?0.06, and 115.46?±?0.47-fold, respectively, relative to the cell-only group. Additionally, hTMSCs within the hybrid-spheroids showed significantly greater osteogenic differentiation under osteogenic media conditions. Taken together, our hybrid-spheroids can be an ideal approach for stem cell expansion and serve as a potential carrier for bone regeneration. Statement of Significance Cells are spatially arranged within extracellular matrix (ECM) and cell/ECM interactions are crucial for cellular functions. Here, we developed a hybrid-spheroid system incorporating engineered ECM prepared from fragmented electrospun fibers to tune stem cell functions. Conventionally prepared cell spheroids with large diameters (>200?μm) is often prone to hypoxia. In contrast, the hybrid-spheroids significantly enhanced viability and proliferation of human turbinate mesenchymal stem cells (hTMSCs) as compared to spheroid prepared from cell only. Under these conditions, the presence of fragmented fibers also improved maintenance of stemness of hTMSCs for longer time cultured in growth media and demonstrated significantly greater osteogenic differentiation under osteogenic media conditions. Thus, the hybrid-spheroids can be used as a delivery carrier for stem cell based therapy or a 3D culture model for in vitro assay.
机译：图形摘要显示省略摘要摘要外细胞外基质（ECM）微环境对于干细胞的活力，茎和分化至关重要。在这项研究中，我们通过使用模拟纤维（FFS）的细胞外基质（ECM）来开发人鼻甲间充质干细胞（HTMSCs）的杂化球状体，以改善HTMSCs的活力和功能。通过部分氨基次氨基溶解（PLLA）纤维板（FS），我们制备平均大小的平均大小的FFS，其涂覆有多碳胺的聚二胺以改善细胞粘附。与碎片纤维混合的杂交球体内的HTMSCs的增殖显着增加，与电池的群体相比显着增加。细胞和碎裂的纤维在结构中存在孔的存在，均匀分布。 LOX-1染色表明，杂交球体改善了细胞活力，这可能是由于通过工程ECM产生的空隙空间增强了氧的运输。透射电子显微镜（TEM）分析证实了杂交 - 球状体内的细胞形成了强的细胞连接和与碎片纤维的触点。在杂交球体中显着上调包括Connexin 43和E-Cadherin的细胞结蛋白的表达在杂交 - 球状体中显着上调16.53Ω·？0.04和28.26？±0.11倍，比仅来自小区的组。类似地，在同一组中，整合蛋白α2，α5和β1的表达显着增强25.72Ω·？0.13,27.48？±0.49和592.78？±0.06倍。另外，在杂交球体中，在杂交球体中显着上调，包括10〜4，±0.06,158.95？±0.47倍，相对于细胞 - 只有群体。另外，在杂交球体内的HTMSCs在成骨介质条件下显示出显着更大的骨质分化。连同，我们的杂交球体可以是干细胞膨胀的理想方法，并用作骨再生的潜在载体。显着性细胞的陈述在细胞外基质（ECM）内，细胞/ ECM相互作用对于细胞功能至关重要。在这里，我们开发了一种掺入由碎片的电纺纤维制备的工程ECM的混合球体系统，以调节干细胞功能。常规制备的具有大直径（>200≤μm）的细胞球体通常易患缺氧。相反，与仅由电池制备的球体相比，杂化 - 球状体显着提高人鼻甲间充质干细胞（HTMSCs）的可存活率和增殖。在这些条件下，碎片纤维的存在还改善了HTMSCs茎干的维持，以便在生长培养基中培养的较长时间，并在成骨介质条件下显示出显着更大的骨质成骨分化。因此，杂交 - 球状体可以用作用于干细胞的治疗或用于体外测定的3D培养模型的输送载体。

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