Biological characteristics of mesenchymal stem cells grown on different topographical nanofibrous poly-L-lactide meshes.
Biological characteristics of mesenchymal stem cells grown on different topographical nanofibrous poly-L-lactide meshes.
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DOI：
10.1166/jbn.2013.1661
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发表时间：
2013-10
影响因子：
2.9
通讯作者：
Jing-xian Zhu;Q. Cai;Xin Zhang;Xiaoqing Hu;La Li;Weiping Wang;Zhenxing Shao;Linghui Dai;Liyuan Cheng;Xiaoping Yang;Chun-yan Zhou;Y. Ao
中科院分区：
文献类型：
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作者：
Jing-xian Zhu;Q. Cai;Xin Zhang;Xiaoqing Hu;La Li;Weiping Wang;Zhenxing Shao;Linghui Dai;Liyuan Cheng;Xiaoping Yang;Chun-yan Zhou;Y. Ao
人工支架的纳米形貌特征对干细胞的生物学特性有复杂的影响。它们影响细胞的粘附、扩散、增殖和分化；然而，我们对这些过程如何在纳米地形线索下发生的了解有限。在本研究中，制备了两种不同纤维排列（完全无纺布和晶格状）的电纺丝纳米纤维网，并将其用于间充质干细胞（MSCs）的体外培养。通过比较与成骨分化相关的特征标记，我们发现随着培养时间的延长，骨桥蛋白（OPN）、骨钙素（OCN）和碱性磷酸酶（ALP）以及相关基因Runx2和Colla基因在晶格状纳米纤维网上的表达水平均高于非织造网。这些结果表明，晶格状纳米纤维网状物激活了MSCs的成骨分化，这是由于纳米纤维定向改变了细胞形态。与纯无纺布纳米纤维网相比，晶格型纳米纤维网具有平行区、磁力线区和无纺布区的组合结构。粘附MSCs后，整合素亚基a5和b1的表达水平上调，激活Ras同源基因家族成员A （RhoA）和细胞外信号调节激酶（ERK）的下游信号通路。当使用特异性抑制剂PD98059和Y27632分别抑制磷酸化ERK和p160 ROCKII活性时，F-actin紊乱，Runx2表达水平下调。因此，我们得出结论，支架纳米形貌可能通过RhoA和ERK信号通路调节MSCs的微环境，促进其成骨分化。这些发现为MSCs在骨组织工程中应用的人工基质的选择提供了有价值的信息。
The nanotopographical features of artificial scaffolds have complex effects on the biological characteristics of stem cells. They influence cell adhesion, spreading, proliferation, and differentiation; however we have limited knowledge on how these processes occur under nanotopographical cues. In this study, two kinds of electrospun nanofibrous meshes with different fiber arrangements (totally non-woven and lattice-like) were fabricated and used for in vitro culture of mesenchymal stem cells (MSCs). By comparing the characteristic marks related to osteogenic differentiation, we found that with prolonged culture time, osteopontin (OPN), osteocalcin (OCN) and alkaline phosphatase (ALP), as well as related genes (Runx2 and Colla genes), were all expressed at higher levels on lattice-like nanofibrous meshes than on non-woven ones. These results indicated that the lattice-like nanofibrous mesh activated the osteogenic differentiation of MSCs owing to changes in cell morphology directed by nanofiber orientations. Compared with pure non-woven nanofibrous meshes, lattice-like ones possessed a combined structure of parallel, magnetic-line-like, and non-woven regions. MSCs adhering onto them had upregulated expression levels of integrin subunits a5 and b1, and activated downstream signaling pathways of Ras homolog gene family member A (RhoA) and extracellular signal-regulated kinase (ERK). When the specific inhibitors PD98059 and Y27632 were used to inhibit phosphorylated ERK and p160 ROCKII activity, respectively, F-actin became disordered and the expression level of Runx2 was downregulated. Thus, we concluded that the scaffold nanotopography may modulate the microenvironment of MSCs and promote their osteogenic differentiation through the RhoA and ERK signaling pathways. These findings provided valuable information on the selection of artificial matrices suitable for MSCs application in bone tissue engineering.