Wu Feng, Chen Huan, Liu Jinyao, Pang Yan
State Key Laboratory of Systems Medicine for Cancer, Shanghai Cancer Institute, Shanghai Key Laboratory for Nucleic Acid Chemistry and Nanomedicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
School of Pharmaceutical Sciences, Shanghai Jiao Tong University, Shanghai, China.
Nat Protoc. 2025 Sep 12. doi: 10.1038/s41596-025-01247-4.
Hydrogels, as 3D cross-linked hydrophilic networks that exhibit favorable flexibility, cargo loading and release abilities and structure and function designability, are desirable for diverse biomedical applications. For in vivo implementation, however, hydrogels often suffer from swelling-weakened mechanical strength, uncontrollable cargo release and complex composition, inevitably hindering further translation. Despite different reported synthetic approaches, the development of a facile yet universal method capable of fabricating hydrogels with dynamically adjustable structure and function remains difficult. Recently, inspired by biological tissues, we have developed a versatile biological membrane hybridization strategy to generate structurally and functionally programmable hydrogels. Specifically, biological membranes are used as a cross-linker to form a cross-linked network through a supramolecular-covalent cascade reaction route. This protocol demonstrates the construction of two biological membrane-hybridized hydrogels, including liposome-hybridized muscle-mimicking hydrogels with swelling-strengthening mechanical behavior and extracellular vesicle-hybridized skin-mimicking hydrogels with enhanced mechanical strength, lubricity, antibacterial activity and immunoactivity. We describe the detailed preparation procedures and characterize the structures and functions of the obtained hydrogels. We also expand the applicability of this biological membrane hybridization strategy to further tune the structure and function of the biomimetic hydrogels by incorporating a second network. This protocol provides a robust preparative platform to develop dual structure- and function-tunable hydrogels for different biomedical applications. Excluding the synthesis of reactive group-functionalized biological membranes, the fabrication of muscle-mimicking hydrogels takes ~3 d, while the construction of skin-mimicking hydrogels takes ~1 d. The implementation of the protocol requires expertise in polymer modification, hydrogel preparation, nanoscale vesicles, surface functionalization and cell culture.
水凝胶作为一种三维交联的亲水性网络，具有良好的柔韧性、载药和释药能力以及结构和功能可设计性，适用于多种生物医学应用。然而，对于体内应用而言，水凝胶常常存在机械强度因溶胀而减弱、药物释放不可控以及成分复杂等问题，不可避免地阻碍了其进一步转化应用。尽管有不同的合成方法报道，但开发一种能够制备结构和功能可动态调节的水凝胶的简便通用方法仍然具有挑战性。最近，受生物组织启发，我们开发了一种通用的生物膜杂交策略来制备结构和功能可编程的水凝胶。具体而言，生物膜用作交联剂，通过超分子 - 共价级联反应路线形成交联网络。本方案展示了两种生物膜杂交水凝胶的构建，包括具有溶胀增强机械性能的脂质体杂交肌肉模拟水凝胶和具有增强机械强度、润滑性、抗菌活性和免疫活性的细胞外囊泡杂交皮肤模拟水凝胶。我们描述了详细的制备过程，并对所得水凝胶的结构和功能进行了表征。我们还通过引入第二个网络，扩展了这种生物膜杂交策略的适用性，以进一步调节仿生水凝胶的结构和功能。该方案提供了一个强大的制备平台，用于开发用于不同生物医学应用的双结构和功能可调水凝胶。除了反应性基团功能化生物膜的合成外，肌肉模拟水凝胶的制备大约需要3天，而皮肤模拟水凝胶的构建大约需要1天。该方案的实施需要聚合物改性、水凝胶制备、纳米级囊泡、表面功能化和细胞培养等方面的专业知识。
