Pelayo-Punzano Gloria, Cuesta Rafael, Calvino José J, Domínguez-Vera José M, López-Haro Miguel, de Vicente Juan, Gálvez Natividad
Department of Inorganic Chemistry, University of Granada, 18071 Granada, Spain.
Department of Organic and Inorganic Chemistry, EPS Linares, University of Jaén, 23700 Linares, Spain.
ACS Appl Mater Interfaces. 2025 Aug 20;17(33):46771-46785. doi: 10.1021/acsami.5c11459. Epub 2025 Aug 5.
Fibrillar protein hydrogels are promising sustainable biomaterials for biomedical applications, but their practical use is often limited by insufficient mechanical strength and stability. To address these challenges, we transformed native proteins into amyloid fibrils (AFs) and incorporated a fibrillar polysaccharide, phytagel (PHY), to engineer interpenetrating polymer network (IPN) hydrogels. Notably, we report for the first time the formation of an amyloid-based hydrogel from apoferritin (APO), with PHY reinforcing the network's mechanical integrity. In situ self-assembly of APO within the PHY matrix yields fully natural, biopolymer-based IPNs. Rheological analyses confirm synergistic interactions between AF and PHY fibers, with the composite hydrogels exhibiting significantly enhanced viscoelastic moduli compared with individual components. The AF-PHY hydrogels also demonstrate excellent self-healing behavior, rapidly restoring their storage modulus after high-strain deformation. A major advancement of this study is the application of deep learning (DL)-based image analysis, using convolutional neural networks, to automate the identification, segmentation, and quantification of fibrillar components in high-resolution scanning electron microscopy images. This AI-driven method enables precise differentiation between AF and PHY fibers and reveals the three-dimensional microarchitecture of the IPN, overcoming key limitations of traditional image analysis. Complementary real-time confocal laser scanning microscopy, with selective fluorescent labeling of protein and polysaccharide components, further validates the IPN structure of the hybrid hydrogels. Our results demonstrate that DL significantly enhances structural characterization and provides insights into gelation processes. This approach sets a new guide for the analysis of complex soft materials and underlines the potential of AF-PHY hydrogels as mechanically robust, self-healing, and fully sustainable biomaterials for biomedical engineering applications.
纤维状蛋白质水凝胶是用于生物医学应用的很有前景的可持续生物材料,但其实际应用常常受到机械强度和稳定性不足的限制。为应对这些挑战,我们将天然蛋白质转化为淀粉样纤维(AFs),并引入一种纤维状多糖——植物凝胶(PHY),以构建互穿聚合物网络(IPN)水凝胶。值得注意的是,我们首次报道了由脱铁铁蛋白(APO)形成基于淀粉样蛋白的水凝胶,其中PHY增强了网络的机械完整性。APO在PHY基质内原位自组装产生完全天然的、基于生物聚合物的IPN。流变学分析证实了AF和PHY纤维之间的协同相互作用,与单个组分相比,复合水凝胶表现出显著增强的粘弹性模量。AF-PHY水凝胶还表现出优异的自愈行为,在高应变变形后能迅速恢复其储能模量。本研究的一个主要进展是应用基于深度学习(DL)的图像分析,使用卷积神经网络,自动识别、分割和量化高分辨率扫描电子显微镜图像中的纤维状组分。这种人工智能驱动的方法能够精确区分AF和PHY纤维,并揭示IPN的三维微结构,克服了传统图像分析的关键局限性。互补的实时共聚焦激光扫描显微镜,通过对蛋白质和多糖组分进行选择性荧光标记,进一步验证了混合水凝胶的IPN结构。我们的结果表明,DL显著增强了结构表征,并为凝胶化过程提供了见解。这种方法为分析复杂软材料设定了新的指导方针,并强调了AF-PHY水凝胶作为用于生物医学工程应用的机械坚固、自愈且完全可持续的生物材料的潜力。
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