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Extended characterization of IL-33/ST2 as a predictor for wound age determination in skin wound tissue samples of humans and mice

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Abstract

Interleukin (IL)-33, an important inflammatory cytokine, is highly expressed in skin wound tissue and serum of humans and mice, and plays an essential role in the process of skin wound healing (SWH) dependent on the IL-33/suppression of tumorigenicity 2 (ST2) pathway. However, whether IL-33 and ST2 themselves, as well as their interaction, can be applied for skin wound age determination in forensic practice remains incompletely characterized. Human skin samples with injured intervals of a few minutes to 24 hours (hs) and mouse skin samples with injured intervals of 1 h to 14 days (ds) were collected. Herein, the results demonstrated that IL-33 and ST2 are increased in the human skin wounds, and that in mice skin wounds, there is an increase over time, with IL-33 expression peaking at 24 hs and 10 ds, and ST2 expression peaking at 12 hs and 7 ds. Notably, the relative quantity of IL-33 and ST2 proteins < 0.35 suggested a wound age of 3 hs; their relative quantity > 1.0 suggested a wound age of 24 hs post-mouse skin wounds. In addition, immunofluorescent staining results showed that IL-33 and ST2 were consistently expressed in the cytoplasm of F4/80-positive macrophages and CD31-positive vascular endothelial cells with or without skin wounds, whereas nuclear localization of IL-33 was absent in α-SMA-positive myofibroblasts with skin wounds. Interestingly, IL-33 administration facilitated the wound area closure by increasing the proliferation of cytokeratin (K) 14 -positive keratinocytes and vimentin-positive fibroblasts. In contrast, treating with its antagonist (i.e., anti-IL-33) or receptor antagonist (e.g., anti-ST2) exacerbated the aforementioned pathological changes. Moreover, treatment with IL-33 combined with anti-IL-33 or anti-ST2 reversed the effect of IL-33 on facilitating skin wound closure, suggesting that IL-33 administration facilitated skin wound closure through the IL-33/ST2 signaling pathway. Collectively, these findings indicate that the detection of IL-33/ST2 might be a reliable biomarker for the determination of skin wound age in forensic practice.

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Data availability

All data generated or analyzed during this study are included in Figs. 1–5. Additional data sets that support the findings of this study are available from the corresponding author upon reasonable request.

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Funding

This work was supported by the National Natural Science Foundation of China (82101972, 81530062, 81971800, 81971163, 82001382, and 82271409), Jiangsu Provincial Natural Science Foundation of China (SBK2020040785), Suzhou Municipal Science and Technology Bureau (SYS2019027), China Postdoctoral Science Foundation (2020M681723), Undergraduate Training Program for Innovation and Entrepreneurship, Soochow University (202110285044Z), a project of invigorating healthcare through science, technology, and education (KJXW2019018), and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

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Authors and Affiliations

  1. Department of Forensic Medicine, School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, 215123, China

    Yuan Gao, Luwei Cai, Yulu Wu, Wenjing Ren, Yirui Song, Heng Xu, Chengliang Luo, Tao Wang & Luyang Tao

  2. Department of Forensic Science, Wenzhou Medical University, Wenzhou, Zhejiang, 325035, China

    Yuan Gao, Luwen Zhu, Youzhuang Wu & Ziguang Lei

  3. Department of Orthopedics, the Affiliated Hospital of Xuzhou Medical University, Xuzhou, 221000, Jiangsu, China

    Dongya Li

  4. Department of Child and Adolescent Healthcare, Children’s Hospital of Soochow University, Suzhou, 215021, Jiangsu, China

    Lili Li

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  1. Yuan Gao
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Contributions

Yuan Gao, Luwei Cai, Ziguang Lei, and Luwen Zhu established the SWH model, and carried out molecular biology experiments; Yuan Gao, Chengliang Luo, and Luyang Tao analyzed the data and wrote the manuscript. Tao Wang, Dongya Li, Youzhuang Wu, and Hen Xu helped collect tissue samples. Yulu Wu, Wenjing Ren, Yirui Song, Dongya Li, and Lili Li participated in data interpretation. Yuan Gao and Chengliang Luo reviewed and edited the manuscript. All authors have contributed significantly to the design of this experiment, participated in the drafting and rigorous review of this manuscript, and endorsed its final version.

Corresponding authors

Correspondence to Ziguang Lei or Luyang Tao.

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Research involving human participants and/or animals

The skin wound tissues of humans and mice involved in this study were obtained with the approval of the Ethics Committee of Soochow University and the Forensic Center of Soochow University.

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Informed consent was obtained from all individual participants included in the study and/ or from legal representatives.

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Ethical approval has been obtained from the Ethics Committee of Soochow University for animal experimentation and human Samples.

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Yuan Gao, Luwei Cai and Dongya Li contributed equally to this work.

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Gao, Y., Cai, L., Li, D. et al. Extended characterization of IL-33/ST2 as a predictor for wound age determination in skin wound tissue samples of humans and mice. Int J Legal Med 137, 1287–1299 (2023). https://doi.org/10.1007/s00414-023-03025-x

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