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Physical dynamic double-network hydrogels as dressings to facilitate tissue repair

Nature Protocols volume 18, pages 3322–3354 (2023)Cite this article

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Abstract

Double-network hydrogels can be tuned to have high mechanical strength, stability, elasticity and bioresponsive properties, which can be combined to create self-healing, adhesive and antibacterial wound dressings. Compared with single-network hydrogel, double-network hydrogel shows stronger mechanical properties and better stability. In comparison with chemical bonds, the cross-linking in double networks makes them more flexible than single-network hydrogels and capable of self-healing following mechanical damage. Here, we present the stepwise synthesis of physical double-network hydrogels where hydrogen bonds and coordination reactions provide self-healing, pH-responsive, tissue-adhesive, antioxidant, photothermal and antibacterial properties, and can be removed on demand. We then explain how to carry out physical, chemical and biological characterizations of the hydrogels for use as wound dressings, yet the double-network hydrogels could also be used in different applications such as tissue engineering scaffolds, cell/drug delivery systems, hemostatic agents or in flexible wearable devices for monitoring physiological and pathological parameters. We also outline how to use the double-network hydrogels in vivo as wound dressings or hemostatic agents. The synthesis of the ureido–pyrimidinone-modified gelatin, catechol-modified polymers and the hydrogels requires 84 h, 48 h and 1 h, respectively, whereas the in vivo assays require 3.5 weeks. The procedure is suitable for users with expertise in biomedical polymer materials.

Key points

  • The approach combines the use of ureido–pyrimidinone hydrogen bonding with metal-coordination interactions between Fe3+ and catechol groups, thereby generating double physical cross-linked hydrogels.

  • The hydrogels have fast self-healing properties, high mechanical strength and can be tuned to respond to conditions such as temperature, pH and light. When used as dressings, the hydrogels facilitate tissue repair in vivo.

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Fig. 1: Schematic diagram of PEGSD/GTU hydrogel preparation.
Fig. 2: Characterization of PEGSD and GTU polymers.
Fig. 3: Properties of the physical double-network hydrogel.
Fig. 4: Self-healing behavior of the PEGSD2/GTU5.0 hydrogel.
Fig. 5: Photothermal antibacterial results of the hydrogel.
Fig. 6: Antioxidant and on-demand removal of the hydrogel.
Fig. 7: Tissue adhesive and biocompatibility of the hydrogels.
Fig. 8: Wound healing using physical double-network hydrogel.
Fig. 9: Tissue adhesives using physical double-network hydrogel.
Fig. 10: The hemostatic effect of the hydrogel hemostatic agent in a rabbit ear artery bleeding model.

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Data availability

The main data discussed in this protocol are available in the supporting primary research paper10.

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Acknowledgements

This work was jointly supported by the National Natural Science Foundation of China (grant numbers 51973172 and 52273149), State Key Laboratory for Mechanical Behavior of Materials, and the World-Class Universities (Disciplines) and the Characteristic Development Guidance Funds for the Central Universities.

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Authors and Affiliations

  1. State Key Laboratory for Mechanical Behavior of Materials, and Frontier Institute of Science and Technology, Xi’an Jiaotong University, Xi’an, China

    Baolin Guo, Yongping Liang & Ruonan Dong

  2. Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research, College of Stomatology, Xi’an Jiaotong University, Xi’an, China

    Baolin Guo

  3. Department of Orthopaedics, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China

    Baolin Guo

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  1. Baolin Guo
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Contributions

B.G. conceived and supervised the project and provided funding. B.G., Y.L. and R.D. conceived and managed the manuscript preparation. Y.L. and B.G. revised the manuscript with input from all authors. All authors read and approved the final manuscript.

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Correspondence to Baolin Guo.

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Key references using this protocol

Zhao, X. et al. Adv. Funct. Mater. 30, 1910748 (2020): https://doi.org/10.1002/adfm.201910748

Zhao, X. et al. Biomaterials 122, 34–47 (2017): https://doi.org/10.1016/j.biomaterials.2017.01.011

Liang, Y. Q. et al. ACS Nano 15, 7078–7093 (2021): https://doi.org/10.1021/acsnano.1c00204

Yu, R. et al. Adv. Healthc. Mater. 11, 2102749 (2022): https://doi.org/10.1002/adhm.202102749

Yu, R. et al. Sci. China Chem. 65, 2238–2251 (2022): https://doi.org/10.1007/s11426-022-1322-5

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Guo, B., Liang, Y. & Dong, R. Physical dynamic double-network hydrogels as dressings to facilitate tissue repair. Nat Protoc 18, 3322–3354 (2023). https://doi.org/10.1038/s41596-023-00878-9

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