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首页> 外文期刊>Acta biomaterialia >Molecular and macro-scale analysis of enzyme-crosslinked silk hydrogels for rational biomaterial design
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Molecular and macro-scale analysis of enzyme-crosslinked silk hydrogels for rational biomaterial design

机译：理性生物材料设计酶交联丝水凝胶的分子与宏观规模分析

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Graphical abstract Display Omitted Abstract Silk fibroin-based hydrogels have exciting applications in tissue engineering and therapeutic molecule delivery; however, their utility is dependent on their diffusive properties. The present study describes a molecular and macro-scale investigation of enzymatically-crosslinked silk fibroin hydrogels, and demonstrates that these systems have tunable crosslink density and diffusivity. We developed a liquid chromatography tandem mass spectroscopy (LC-MS/MS) method to assess the quantity and order of covalent tyrosine crosslinks in the hydrogels. This analysis revealed between 28 and 56% conversion of tyrosine to dityrosine, which was dependent on the silk concentration and reactant concentration. The crosslink density was then correlated with storage modulus, revealing that both crosslinking and protein concentration influenced the mechanical properties of the hydrogels. The diffusive properties of the bulk material were studied by fluorescence recovery after photobleaching (FRAP), which revealed a non-linear relationship between silk concentration and diffusivity. As a result of this work, a model for synthesizing hydrogels with known crosslink densities and diffusive properties has been established, enabling the rational design of silk hydrogels for biomedical applications. Statement of Significance Hydrogels from naturally-derived silk polymers offer versitile opportunities in the biomedical field, however, their design has largely been an empirical process. We present a fundamental study of the crosslink density, storage modulus, and diffusion behavior of enzymatically-crosslinked silk hydrogels to better inform scaffold design. These studies revealed unexpected non-linear trends in the crosslink density and diffusivity of silk hydrogels with respect to protein concentration and crosslink reagent concentration. This work demonstrates the tunable diffusivity and crosslinking in silk fibroin hydrogels, and enables the rational design of biomaterials. Further, the characterization methods presented have applications for other materials with dityrosine crosslinks, which are found in nature as post-translational modificaitons, as well as in engineered matrices such as tyramine-substituted hyaluronic acid and recombinant resilin.
机译：图形抽象显示省略了抽象的丝绸纤维素的水凝胶在组织工程和治疗分子递送中具有激动的应用;然而，他们的效用取决于他们的扩散特性。本研究描述了酶促交联的丝纤维素水凝胶的分子和宏观调查，并证明这些系统具有可调谐的交联密度和扩散性。我们开发了一种液相色谱串联质谱（LC-MS / MS）方法，以评估水凝胶中共价酪氨酸交联的数量和顺序。该分析显示出28％至56％的酪氨酸转化为酪氨酸，这取决于丝浓度和反应物浓度。然后与储存模量相关联的交联密度，揭示交联和蛋白质浓度两种水凝胶的机械性能。通过光漂白（FRAP）之后通过荧光回收研究了本体材料的扩散性质，这揭示了丝浓度和扩散率之间的非线性关系。由于这项工作，已经建立了一种用已知的交联密度合成水凝胶和扩散特性的模型，实现了丝绸水凝胶的理性设计，用于生物医学应用。自然衍生的丝绸聚合物的意义水凝胶陈述提供了生物医学领域的多种机会，然而，他们的设计在很大程度上是一个实证过程。我们介绍了酶交联丝水凝胶的交联密度，储存模量和扩散行为的基本研究，以更好地通知支架设计。这些研究揭示了与蛋白质浓度和交联试剂浓度的丝水凝胶交联密度和扩散性的意外的非线性趋势。这项工作证明了丝纤维素水凝胶中的可调谐扩散性和交联，并实现了生物材料的合理设计。此外，所呈现的表征方法具有与双核交联的其他材料的应用，其作为翻译后的修饰，以及在工程化基质中，如酪胺取代的透明质酸和重组resilin。

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