Biomaterials:组织工程肌腱组织蛋白质组学分析获进展

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      近日,中科院大连化学物理研究所生物分离分析新材料与新技术课题组(1809组)邹汉法研究员和上海交通大学刘伟教授合作,将差异蛋白质组学分析策略成功地应用于组织工程肌腱组织分子机制的研究中,在蛋白质水平上探索了机械应力对组织工程肌腱构建的影响。相关研究结果发表在《生物材料》(Biomaterials)杂志上。

      组织工程肌腱的形成涉及多种分子相互作用的复杂网络,传统的生物学方法如RT-PCR和Western blot技术只能针对某几个基因或蛋白分子,不足以完全解释其形成机制。蛋白质组学分析平台实现了蛋白的大规模鉴定,差异蛋白质组学分析发现了包括细胞外基质形成、胞内信号传导、细胞骨架构建以及炎症应答等细胞功能在力学刺激前后的显著变化,实现了与组织工程成腱过程相关的目标蛋白筛选。该工作不仅为研究机械应力在肌腱形成过程中的作用机制提供参考,而且对组织工程组织发生、发展过程的分子机制也有重要的指导意义生物谷推荐更多原文:

      Biomaterials     DOI:10.1016/j.biomaterials.2011.02.033

      A proteomic analysis of engineered tendon formation under dynamic mechanical loading in vitro

      Yongkang Jianga, 1, Hongwei Liub, 1, Hong Lic, d, Fangjun Wangb, Kai Chengb, Guangdong Zhoua, c, Wenjie Zhanga, c, Mingliang Yeb, Yinlin Caoa, c, Wei Liua, c, , and Hanfa Zoub

      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 Wnt signal pathway) mediated other biological processes.

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