Proteomic analysis of a decellularized human vocal fold mucosa scaffold using 2D electrophoresis and high-resolution mass spectrometry.
Proteomic analysis of a decellularized human vocal fold mucosa scaffold using 2D electrophoresis and high-resolution mass spectrometry.
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DOI：
10.1016/j.biomaterials.2012.09.050
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发表时间：
2013-01
期刊：
影响因子：
14
通讯作者：
Frey, Brian L.
中科院分区：
文献类型：
--
作者：
Welham, Nathan V.;Chang, Zhen;Smith, Lloyd M.;Frey, Brian L.
用于组织工程的天然生物支架通常是通过组织和器官的脱细胞产生的。尽管在临床前和临床上取得了一些成功，但体内支架重塑和功能结果仍然存在差异，这可能是由于未识别的生物活性分子对支架-宿主相互作用的影响。在这里，我们使用二维电泳和基于高分辨率质谱的蛋白质组分析来评估脱细胞效果并鉴定人类声带粘膜模型中潜在的生物活性蛋白质残留物。我们注意到脱细胞后蛋白质组、磷酸蛋白质组和 O-糖蛋白质组的消耗，并在脱细胞支架内鉴定了超过 200 种独特的蛋白质种类。基于基因本体的富集分析揭示了与细胞外基质组装、组织、形态和模式相关的一组主要的功能相关本体术语，这与为以后的细胞播种和渗透保留组织特异性生态位相一致。尽管组织学和免疫组织化学数据表明完全脱细胞化，但我们进一步确定了与生物活性（其中一些是抗原性）细胞蛋白相关的本体术语子集。这些发现证明了基于质谱的蛋白质组学在识别可能导致宿主对脱细胞支架衍生的工程组织反应变化的物质方面的价值。这项工作对于从任何组织或器官制造生物支架以及预测和监测体内支架-宿主相互作用具有重要意义。
Natural biologic scaffolds for tissue engineering are commonly generated by decellularization of tissues and organs. Despite some preclinical and clinical success, in vivo scaffold remodeling and functional outcomes remain variable, presumably due to the influence of unidentified bioactive molecules on the scaffold-host interaction. Here, we used 2D electrophoresis and high-resolution mass spectrometry-based proteomic analyses to evaluate decellularization effectiveness and identify potentially bioactive protein remnants in a human vocal fold mucosa model. We noted proteome, phosphoproteome and O-glycoproteome depletion post-decellularization, and identified >200 unique protein species within the decellularized scaffold. Gene ontology-based enrichment analysis revealed a dominant set of functionally-related ontology terms associated with extracellular matrix assembly, organization, morphology and patterning, consistent with preservation of a tissue-specific niche for later cell seeding and infiltration. We further identified a subset of ontology terms associated with bioactive (some of which are antigenic) cellular proteins, despite histological and immunohistochemical data indicating complete decellularization. These findings demonstrate the value of mass spectrometry-based proteomics in identifying agents potentially responsible for variation in host response to engineered tissues derived from decellularized scaffolds. This work has implications for the manufacturing of biologic scaffolds from any tissue or organ, as well as for prediction and monitoring of the scaffold-host interaction in vivo.
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影响因子：
14
作者：
Brown, Bryan N.;Barnes, Christopher A.;Kasick, Rena T.;Michel, Roger;Gilbert, Thomas W.;Beer-Stolz, Donna;Castner, David G.;Ratner, BuddyD.;Badylak, Stephen F.
通讯作者：
Badylak, Stephen F.
影响因子：
82.9
作者：
Ott, Harald C.;Clippinger, Ben;Vacanti, Joseph P.
通讯作者：
Vacanti, Joseph P.
影响因子：
14
作者：
Daly, K. A.;Liu, S.;Badylak, S. F.
通讯作者：
Badylak, S. F.
DOI：
10.1126/science.1189345
发表时间：
2010-07-30
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Petersen TH;Calle EA;Zhao L;Lee EJ;Gui L;Raredon MB;Gavrilov K;Yi T;Zhuang ZW;Breuer C;Herzog E;Niklason LE
通讯作者：
Niklason LE
影响因子：
14
作者：
Brown, Bryan N.;Valentin, Jolene E.;Stewart-Akers, Ann M.;McCabe, George P.;Badylak, Stephen F.
通讯作者：
Badylak, Stephen F.