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A tunable protein-based scaffold for the study of central nervous system regeneration.

机译：一种基于蛋白质的可调节支架，用于研究中枢神经系统的再生。

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Central nervous system (CNS) injuries pose a significant and potentially debilitating health problem in society today and, to date, no successful clinical repair strategies have been advanced. The development of effective treatments is severely hindered by the quick formation of a complex, inhibitory scar at the site of CNS injury. This scar both physically blocks and chemically suppresses nerve regeneration. It has been hypothesized that combinatorial approaches involving biomaterial scaffolds, cell transplantation, and pro-survival factors, which provide a more permissive growth environment, have the highest chance of stimulating regeneration. The work completed in this thesis focuses on the design and characterization of a biomimetic hydrogel scaffold constructed from chemically crosslinked recombinant proteins. This protein-based scaffold has been designed to offer a flexible platform for the systematic optimization of key scaffold design parameters, such as mechanical strength, degradation, cellular interaction, molecule delivery, and topography. Specifically, a collection of proteins containing sequences previously shown to enhance cell adhesion, to promote neurite extension, and to exhibit varying susceptibility to cleavage by neurite-secreted proteases were synthesized to serve as the polymer backbone for the scaffold. Experiments were conducted to analyze the capacity of scaffolds, constructed from single proteins or mixtures of proteins, to independently control cell behavior, scaffold degradation properties, and scaffold mechanical properties based upon differences in the primary protein sequence and crosslinking conditions. In addition, composite scaffolds constructed by layered spatial deposition of chemically crosslinked, protease-degradable proteins were applied to the formation of dynamic internal, three-dimensional scaffold patterns that can be directly coupled to molecule delivery. Overall, this work demonstrates the tunable and bio-functional nature of these hydrogels and sets the framework for future studies into the development of effective protein-engineered scaffolds for CNS regeneration.
机译：中枢神经系统（CNS）损伤在当今社会中构成了严重的且可能使人衰弱的健康问题，并且迄今为止，尚未提出成功的临床修复策略。在CNS损伤部位快速形成复杂的抑制性瘢痕严重阻碍了有效治疗的发展。这种疤痕在物理上既阻塞又化学抑制神经再生。据推测，涉及生物材料支架，细胞移植和促存活因素的组合方法，提供了更为宽松的生长环境，具有最大的刺激再生机会。本论文完成的工作集中于由化学交联的重组蛋白构建的仿生水凝胶支架的设计和表征。这款基于蛋白质的支架旨在为关键支架设计参数（例如机械强度，降解，细胞相互作用，分子递送和形貌）的系统优化提供灵活的平台。具体地，合成了包含先前显示出可增强细胞粘附，促进神经突延伸以及表现出对神经突分泌的蛋白酶裂解的不同敏感性的序列的蛋白质集合，以用作支架的聚合物主链。进行实验以分析由单一蛋白质或蛋白质混合物构成的支架根据一级蛋白质序列和交联条件的差异独立控制细胞行为，支架降解特性和支架机械特性的能力。另外，通过化学交联的，可蛋白酶降解的蛋白质的分层空间沉积而构建的复合支架被应用于动态内部三维支架模式的形成，该模式可以直接与分子递送耦合。总的来说，这项工作证明了这些水凝胶的可调性和生物功能性质，并为开发有效的蛋白质工程支架用于中枢神经系统再生的未来研究奠定了框架。

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