Designing an extracellular matrix mimic by biofunctionalization of polymeric scaffolds is a popular strategy and extremely crucial for facilitating the interactions between cells and the matrix. To this direction, supramolecular gels are gaining exponential attention over the last few years, owing to their potential biocompatibility and biodegradability. In spite of diverse biological roles of native laminin, the bioactivities of self-assembling laminin-derived short peptides were less explored. In this work, we have explored the minimalist design to develop hydrogel scaffolds based on IKVAV and YIGSR peptides individually and their composite matrix, which can provide structurally and functionally relevant materials for tissue engineering. Till date, composite supramolecular gels solely made up of self-assembling IKVAV and YIGSR peptides have never been reported. Such composite gels can be a closer mimic of natural laminin protein, which could mimic the essential functions of the short peptide fragments present on different chains of the extracellular matrix protein, laminin. Interestingly, we used a unique strategy of simple mixing of the two laminin mimetic peptides, which tend to induce coassembly with a self-sorted nanofibrous network with relatively enhanced mechanical strength. The physicochemical properties of the biofunctional hydrogels were studied using different microscopic, spectroscopic, and rheology techniques. To assess the bioactivity of laminin-derived scaffolds in controlling neuronal cell growth, its biocompatibility, cellular growth, and proliferation were quantified using C6 glial cells and SHSY5Y neuroblastoma cells. The live/dead staining further confirmed the adhesion and proliferation of the cells. A significant increase in neurite length provides clear evidence on mimicking the neurite extension function of native laminin protein by its short derivatives. Interestingly, similar β-III tubulin expression and cell cycle phases were observed, in comparison to control, which indicated normal cellular functioning of the cells cultured over short laminin hydrogel scaffolds. All bioassays suggested that Fmoc YIGSR promotes growth of neural cells to a greater extent and maintains healthier morphology, in comparison to hydrophobic Fmoc IKVAV, owing to the entangled longer fibrous network formed by YIGSR peptide. It is expected that thinner long fibers provide a more uniform surface and are more supportive for cell adhesion in comparison to hydrophobic, shorter fibers IKVAV peptide. However, in composite gels, the detrimental effect of hydrophobic IKVAV peptide could be reduced and better adhesion and proliferation could be achieved along with enhanced cell survival. These observations demonstrate the high potential of the laminin-derived hydrogels in tissue engineering and neuronal stem cell differentiation in future.

中文翻译：

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通过复合层粘连蛋白超分子水凝胶控制神经元细胞的生长。

通过聚合物支架的生物功能化设计模拟细胞外基质是一种流行的策略，对于促进细胞与基质之间的相互作用至关重要。在这个方向上，超分子凝胶由于其潜在的生物相容性和生物降解性，在过去几年中获得了广泛的关注。尽管天然层粘连蛋白具有多种生物学作用，但自组装层粘连蛋白衍生的短肽的生物活性却很少被探索。在这项工作中，我们探索了极简设计，以分别开发基于IKVAV和YIGSR肽及其复合基质的水凝胶支架，该支架可为组织工程提供结构和功能上相关的材料。直到日期 仅由自组装IKVAV和YIGSR肽组成的复合超分子凝胶尚未见报道。这样的复合凝胶可以更像天然层粘连蛋白，它可以模仿存在于细胞外基质蛋白层粘连蛋白的不同链上的短肽片段的基本功能。有趣的是，我们使用了一种简单的策略，将两种层粘连蛋白模拟肽简单混合，这倾向于诱导与自选纳米纤维网络的共组装，并具有相对增强的机械强度。使用不同的显微镜，光谱学和流变学技术研究了生物功能水凝胶的物理化学性质。为了评估层粘连蛋白支架在控制神经元细胞生长，其生物相容性，细胞生长中的生物活性，使用C6神经胶质细胞和SHSY5Y神经母细胞瘤细胞对增殖和增殖进行定​​量。活/死染色进一步证实了细胞的粘附和增殖。神经突长度的显着增加提供了通过其短衍生物模拟天然层粘连蛋白的神经突延伸功能的明确证据。有趣的是，与对照相比，观察到相似的β-III微管蛋白表达和细胞周期阶段，表明在短层粘连蛋白水凝胶支架上培养的细胞正常细胞功能。所有生物测定均表明，与疏水性Fmoc IKVAV相比，由于YIGSR肽形成的更长的纤维网络缠结，Fmoc YIGSR可以更大程度地促进神经细胞的生长并保持更健康的形态。与疏水性较短的短纤维IKVAV肽相比，预期较细的长纤维可提供更均匀的表面并更支持细胞粘附。然而，在复合凝胶中，疏水性IKVAV肽的有害作用可以降低，更好的粘附和增殖以及增强的细胞存活率可以实现。这些观察结果表明，层粘连蛋白衍生的水凝胶在组织工程和神经元干细胞分化方面具有很高的潜力。