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Generating structurally and functionally programmable hydrogels by biological membrane hybridization

Nature Protocols (2025)Cite this article

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Abstract

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.

Key points

  • This protocol describes a versatile biological membrane hybridization strategy that uses biological membranes as cross-linkers to generate structurally and functionally programmable hydrogels through a supramolecular-covalent cascade reaction procedure.

  • Compared to conventional cross-linking strategies, this protocol shows flexibility and versatility to generate biomimetic hydrogels, such as muscle-mimicking hydrogels with swelling-strengthening mechanical behavior and skin-mimicking hydrogels with enhanced mechanical strength, lubricity, antibacterial activity and immunoactivity.

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Fig. 1: Schematic illustration of the biological membrane–hybridization strategy to generate structurally and functionally programmable hydrogels.
Fig. 2: Schematic illustration of the preparation, functional evaluation and versatility verification of biological membrane–hybridized hydrogels.
Fig. 3: Characterization of ALip, NALip and SSHs.
Fig. 4: Swelling-strengthening behavior of SSHs in vitro and in vivo.
Fig. 5: Versatility of SSHs.
Fig. 6: Characterization of OMVs, OMV-AM and SFSHs.
Fig. 7: Enhanced mechanical strength, antibacterial ability and immunological activity of SFSHs.
Fig. 8: Versatility of SFSHs.

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Data availability

The main data discussed in this protocol are available in the supporting primary research papers (refs. 38,39). Source data are provided with this paper.

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Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (22425505 to J.L., 22375127 to Y.P., 22475131 to F.W. and 22305152 to H.C.), the Shanghai Rising-Star Program (23QA1408600 to F.W.) and the Innovative Research Team of High-Level Local Universities in Shanghai (SHSMU-ZDCX20210700 to Y.P.).

Author information

Authors and Affiliations

  1. 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

    Feng Wu & Jinyao Liu

  2. School of Pharmaceutical Sciences, Shanghai Jiao Tong University, Shanghai, China

    Huan Chen & Yan Pang

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  1. Feng Wu
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  2. Huan Chen
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  3. Jinyao Liu
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Contributions

Y.P., J.L. and F.W. conceived and managed the manuscript preparation. H.C. helped with the analysis of the results. Y.P., J.L. and F.W. revised the manuscript with input from all authors. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Jinyao Liu or Yan Pang.

Ethics declarations

Competing interests

J.L. and F.W. have applied for a patent (202311812159.9) from the State Intellectual Property Office (China). The authors declare that they have no competing interests.

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Nature Protocols thanks Junjie Deng and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Key references

Wu, F. et al. Nat. Commun. 11, 4502 (2020): https://doi.org/10.1038/s41467-020-18308-9

Wu, F. et al. Nat. Commun. 15, 802 (2024): https://doi.org/10.1038/s41467-024-45006-7

Supplementary information

Source data

Source Data Figs. 3–8

Subsheet: Fig. 3a–g. Subsheet: Fig. 4a–j. Subsheet: Fig. 5a–e. Subsheet: Fig. 6a–i. Subsheet: Fig. 7a–n. Subsheet: Fig. 8a–l

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Wu, F., Chen, H., Liu, J. et al. Generating structurally and functionally programmable hydrogels by biological membrane hybridization. Nat Protoc (2025). https://doi.org/10.1038/s41596-025-01247-4

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