A proteomic analysis of engineered tendon formation under dynamic mechanical loading in vitro
A proteomic analysis of engineered tendon formation under dynamic mechanical loading in vitro
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体外动态机械载荷下工程肌腱形成的蛋白质组学分析
DOI：
10.1016/j.biomaterials.2011.02.033
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发表时间：
2011-06-01
期刊：
影响因子：
14
通讯作者：
Zou, Hanfa
中科院分区：
文献类型：
--
作者：
Jiang, Yongkang;Liu, Hongwei;Zou, Hanfa
先前的研究已经证明了机械载荷对体外肌腱工程的有益作用。为了了解机制，人肌腱细胞和聚乙醇酸长纤维用于体外肌腱工程在生物反应器系统中进行12周的动态加载和不加载。工程化的新肌腱进行蛋白质组学分析，使用质谱沿着与鸟枪策略。正如预期的那样，机械负荷导致更成熟的肌腱组织，其特征在于更坚固的组织质地和密集沉积的基质，这些基质以高度紧凑的方式形成纵向排列的胶原纤维。相比之下，未加载的新肌腱显示松散和沉积较少的矩阵在一个相对较不有组织的模式。从两组组织中分离的蛋白质具有相似的等电点和分子量分布，表明组织标本的相似性和可比性。此外，蛋白质组学分析表明，共758个蛋白质被确定从这两个群体中的194和177蛋白质独特的负载和非负载肌腱，分别。加载和非加载肌腱的比较显示195个显著上调的蛋白和189个显著下调的蛋白。差异表达蛋白大致可分为细胞外基质、细胞内信号、细胞骨架和炎症反应等几类。其中，显著上调的胶原I和VI、MMP-14、WNT 5A、微丝分子和一些炎性因子表明，这种特殊生物学现象的可能机制可能涉及腱特异性基质的产生增加，胶原和其他基质分子的交联增强，组织成熟和机械转导（包括非经典Writ信号通路）介导的其他生物学过程的适当基质重塑。(C)2011爱思唯尔有限公司保留所有权利。
Previous studies have demonstrated the beneficial effect of mechanical loading on in vitro tendon engineering. To understand the mechanism, human tenocytes and polyglycolic acid long fibers were used for in vitro tendon engineering in a bioreactor system for 12 weeks with and without dynamic loading. The engineered neo-tendons were subjected to proteomic analysis using mass spectrometry along with shotgun strategy. As expected, mechanical loading resulted in a more mature tendon tissue characterized by a firmer tissue texture and densely deposited matrices which formed longitudinally aligned collagen fibers in a highly compact fashion. In contrast, non-loaded neo-tendon revealed loosely and less deposited matrices in a relatively less organized pattern. Proteins isolated from two groups of tissues exhibited similar distribution of isoeletric point and molecular weight indicating the similarity and comparability of the tissue specimens. Further, proteomic analysis showed that total 758 proteins were identified from both groups with 194 and 177 proteins uniquely presented in loaded and non-loaded tendons, respectively. Comparison of loaded and non-loaded tendons revealed 195 significantly up-regulated proteins and 189 significantly down-regulated proteins. The differentially expressed proteins could generally be classified into the categories of extracellular matrix, intra-cellular signaling, cytoskeleton and inflammatory response. Among them, significantly up-regulated collagens I and VI, MMP-14, WNT5A, microfilament molecules and some inflammatory factors suggest that the possible mechanism for this particular biological phenomenon may involve increased production of tendon specific matrices, enhanced cross-link of collagens and other matrix molecules, proper matrix remodeling for tissue maturation and mechanotransduction (including non-canonical Writ signal pathway) mediated other biological processes. (C) 2011 Elsevier Ltd. All rights reserved.