Preparation and mechanism study of hydrogen bond induced enhanced composited gelatin microsphere probe,International Journal of Biological Macromolecules

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Preparation and mechanism study of hydrogen bond induced enhanced composited gelatin microsphere probe
International Journal of Biological Macromolecules ( IF 7.7 ) Pub Date : 2024-03-11 , DOI: 10.1016/j.ijbiomac.2024.130752
Xiaoyong Tian ₁ , Kezhen Zhang ₁ , Yu Zhang ₂ , Nan Wang ₂ , Huaping Wang ₁ , Hongyao Xu ₁ , Shanyi Guang ₂

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Fluorescent probes offer rapid and efficient detection of metal ions. However, their properties, including high biotoxicity and low detection limits, often limit their utility in biological systems. In this study, we used a microfluidic approach to fabricate photocrosslinked gelatin microspheres with a micropore, providing a straightforward method for loading fluorescent probes into these microspheres based on the adsorption effect and hydrogen bonding interaction. The gelatin microsphere loaded probes, GelMA/TPA-DAP and GelMA/TPA-ISO-HNO were designed and obtained. The results show that these probes exhibit obviously low biotoxicity compared to the original molecular probes TPA-DAP and TPA-ISO-HNO. Simultaneously, it is found that GelMA/TPA-DAP and GelMA/TPA-ISO-HNO have better detection sensitivity, the detection limits are 35.4 nM for Cu, 16.5 nM for Co and 20.5 nM for Ni for GelMA/TPA-DAP probe. Compared to the original TPA-DAP they are improved by 37.2 %, 26.3 % and 22.6 % respectively. The corresponding coordination constants were 10.8 × 10, 4.11×10 and 6.04×10, which is larger than homologous TPA-DAP. Similar results were also verified in the GelMA/TPA-ISO-HNO probe. The mechanism was investigated in detail by theoretical simulations and advanced spectral analysis. The density functional theory (DFT) simulations show that the probes are anchored inside the microspheres and the molecular structure is modified due to the hydrogen bonding interaction between the microsphere and the molecular probe, which makes GelMA/TPA-DAP exhibit stronger coordination capacity with metal ions than homologous TPA-DAP. In addition, the adsorption effect also provided some synergistic enhancement contribution. Meanwhile, cellular experiments have also shown that the composite microspheres can improve the biocompatibility of the probe and will provide a wider range of applications towards bioassay. This simple and effective method will provide a convenient way to improve the performance of fluorescent probes and their biological applications.

中文翻译：

氢键诱导增强复合明胶微球探针的制备及机理研究

荧光探针可快速有效地检测金属离子。然而，它们的特性，包括高生物毒性和低检测限，通常限制了它们在生物系统中的应用。在这项研究中，我们使用微流体方法制备了具有微孔的光交联明胶微球，提供了一种基于吸附效应和氢键相互作用将荧光探针加载到这些微球中的简单方法。设计并制备了明胶微球负载探针GelMA/TPA-DAP和GelMA/TPA-ISO-HNO。结果表明，与原始分子探针TPA-DAP和TPA-ISO-HNO相比，这些探针表现出明显较低的生物毒性。同时发现GelMA/TPA-DAP和GelMA/TPA-ISO-HNO具有更好的检测灵敏度，GelMA/TPA-DAP探针的检测限为Cu：35.4 nM，Co：16.5 nM，Ni：20.5 nM。与原始TPA-DAP相比，它们分别提高了37.2%、26.3%和22.6%。相应的配位常数分别为10.8×10、4.11×10和6.04×10，比同源的TPA-DAP大。类似的结果也在 GelMA/TPA-ISO-HNO 探针中得到验证。通过理论模拟和先进的光谱分析对该机制进行了详细研究。密度泛函理论（DFT）模拟表明，探针锚定在微球内部，由于微球与分子探针之间的氢键相互作用，分子结构发生改变，使得GelMA/TPA-DAP表现出更强的与金属的配位能力离子比同源的TPA-DAP。此外，吸附效应还提供了一些协同增强的贡献。同时，细胞实验也表明复合微球可以提高探针的生物相容性，将为生物测定提供更广泛的应用。这种简单有效的方法将为提高荧光探针的性能及其生物学应用提供便捷的途径。

更新日期：2024-03-11

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