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首页> 外文期刊>Biomaterials Science >Design of thiol-ene photoclick hydrogels using facile techniques for cell culture applications
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Design of thiol-ene photoclick hydrogels using facile techniques for cell culture applications

机译：使用方便的技术设计用于细胞培养的硫醇烯光点击水凝胶

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Thiol-ene 'click' chemistries have been widely used in biomaterials applications, including drug delivery, tissue engineering, and controlled cell culture, owing to their rapid, cytocompatible, and often orthogonal reactivity. In particular, hydrogel-based biomaterials formed by photoinitiated thiol-ene reactions afford spatiotemporal control over the biochemical and biomechanical properties of the network for creating synthetic materials that mimic the extracellular matrix or enable controlled drug release. However, the use of charged peptides functionalized with cysteines, which can form disulfides prior to reaction, and vinyl monomers that require multistep syntheses and contain ester bonds, may lead to undesired inhomogeneity or degradation under cell culture conditions. Here, we designed a thiol-ene hydrogel formed by the reaction of allyloxycarbonyl-functionalized peptides and thiol-functionalized poly(ethylene glycol). Hydrogels were polymerized by free radical initiation under cytocompatible doses of long wavelength ultraviolet light in the presence of water-soluble photoinitiators (lithium acylphosphinate, LAP, and 2-hydroxy-1-[4-(2-hydroxyethoxy)phenyl]-2-methyl-1-propanone, Irgacure 2959). Mechanical properties of these hydrogels were controlled by varying the monomer concentration to mimic a range of soft tissue environments, and hydrogel stability in cell culture medium was observed over weeks. Patterns of biochemical cues were created within the hydrogels post-formation and confirmed through the incorporation of fluorescently-labeled peptides and Ellman's assay to detect free thiols. Human mesenchymal stem cells remained viable after encapsulation and subsequent photopatterning, demonstrating the utility of the monomers and hydrogels for three-dimensional cell culture. This facile approach enables the formation and characterization of hydrogels with well-defined, spatially-specific properties and expands the suite of monomers available for three-dimensional cell culture and other biological applications.
机译：硫醇-“点击”化学由于其快速，细胞相容性和通常正交的反应性，已被广泛用于生物材料应用中，包括药物输送，组织工程和受控细胞培养。特别地，通过光引发的硫醇-烯反应形成的基于水凝胶的生物材料提供了对网络的生化和生物力学特性的时空控制，以产生模仿细胞外基质或使药物释放受控的合成材料。但是，使用半胱氨酸官能化的带电肽会在反应前形成二硫化物，而需要多步合成并含有酯键的乙烯基单体在细胞培养条件下可能会导致不良的不均一性或降解。在这里，我们设计了由烯丙氧基羰基官能化的肽与硫醇官能化的聚乙二醇反应形成的硫醇烯水凝胶。在水溶性光引发剂（酰基次膦酸锂，LAP和2-羟基-1- [4-（2-羟基乙氧基）苯基] -2-甲基）存在下，在细胞相容剂量的长波长紫外光下，通过自由基引发聚合水凝胶-1-丙酮，Irgacure 2959）。通过改变单体浓度以模拟一系列软组织环境来控制这些水凝胶的机械性能，并在数周内观察到了细胞培养基中水凝胶的稳定性。在水凝胶形成后会产生生化线索的模式，并通过掺入荧光标记的肽和通过Ellman's检测游离硫醇的方法进行确认。人间充质干细胞在包封和随后的光图案化之后仍然保持活力，证明了单体和水凝胶在三维细胞培养中的效用。这种简便的方法能够形成和表征具有明确定义的，空间上特定的特性的水凝胶，并扩展了可用于三维细胞培养和其他生物学应用的单体套件。

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