Abstract
Diabetic foot ulcers often become infected, leading to treatment complications and increased risk of loss of limb. Therapeutics to manage infection and simultaneously promote healing are needed. Here we report on the development of a Janus liposozyme that treats infections and promotes wound closure and re-epithelialization. The Janus liposozyme consists of liposome-like selenoenzymes for reactive oxygen species (ROS) scavenging to restore tissue redox and immune homeostasis. The liposozymes are used to encapsulate photosensitizers for photodynamic therapy of infections. We demonstrate application in methicillin-resistant Staphylococcus aureus-infected diabetic wounds showing high ROS levels for antibacterial function from the photosensitizer and nanozyme ROS scavenging from the liposozyme to restore redox and immune homeostasis. We demonstrate that the liposozyme can directly regulate macrophage polarization and induce a pro-regenerative response. By employing single-cell RNA sequencing, T cell-deficient Rag1−/− mice and skin-infiltrated immune cell analysis, we further reveal that IL-17-producing γδ T cells are critical for mediating M1/M2 macrophage transition. Manipulating the local immune homeostasis using the liposozyme is shown to be effective for skin wound repair and tissue regeneration in mice and mini pigs.
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Data availability
All generated or analysed data supporting the findings of this study are available within the paper and its Supplementary Information. The RNA-seq data are available from the Gene Expression Omnibus (GEO) database, with accession number GSE238152. The single-cell RNA-seq data are available from the GEO database, with accession number GSE253098. All raw data from this study are available from the corresponding authors upon request. Source data are provided with this paper.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China project (81921004, D.K.; 82241218 and 31972896, S.Z.; 82372140, C.Z.), National Key Research and Development Program of China (2021YFA1201103, S.Z.) and Tianjin Natural Science Foundation (22JCZDJC00180, C.Z.; 22JCYBJC00460, D.L.) and we acknowledge the financial support from the Fundamental Research Funds for the Central Universities (63231049, D.L.; 63213080, C.Z.) and Fundamental Research Funds for Institute of Transplantation Medicine of Nankai University (NKTM2023003, S.Z.; NKTM2023004, C.Z.). We thank PETCC, the Global Pets’ Cell Resource Center, for kindly providing us with cell lines and medium for testing. We also thank the Flow Cytometry Core Facility, Microscopy Platform, Bioinformatics Platform and Mass Spectrometry Platform at the College of Life Sciences, Nankai University, for supporting our work.
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Contributions
C.Z. and S.Z. designed the study. C.Z. conceived the idea and developed the materials and method for the Janus liposozyme. T.P. and X.M. synthesized seleno-phospholipid (Se-DOPE). Y.Z. and J.Q. designed and synthesized TDTM. T.W. and T.P. designed the in vitro and in vivo experiment. T.W., T.P., Y.G., X.M., J.Q. and Y.Z. characterized the Janus liposozyme in vitro. T.W., Y.G. and W.W. evaluated the antibacterial effect. T.W., X.M. and Y.G. designed and conducted the in vivo diabetic wound healing experiment and analysis in WT mice and Bama mini pigs. T.W., X.P., R.G., M.Z. and D.L. designed and conducted the in vivo diabetic wound healing experiment and analysis in genetic ablation of T cells in mice. X.P., R.G., F.K. and M.H. completed the flow cytometry experiment and analysis. T.W. and D.Z. performed histology assessment, immunofluorescence and RNA-seq analysis. X.P. and M.Z. conducted single-cell RNA-seq experiments. X.P., F.D., M.Z. and S.Z. contributed to single-cell RNA-seq analysis. T.W., X.P., M.Z., L.Z. and C.Z. prepared the figures. T.W., X.P., M.Z., D.K., S.Z. and C.Z. wrote the manuscript with inputs from all authors.
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C.Z., T.P., X.M., T.W. and S.Z. are the inventors of a patent application (application no. 2024102459627) that covers the synthesis and antioxidative function of seleno-phospholipids. The other authors declare no competing interests.
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Extended data
Extended Data Fig. 1 In vivo wound healing efficacy on db/db mice infected by MRSA.
a, Representative images of infected diabetic wounds of db/db mice in different treatment groups. Inside the green dashed line is the initial wound area. Scale bars, 2 mm. b, In vivo wound closure rates of db/db mice in different treatment groups. n = 5 biologically independent samples. c, Representative H&E staining images and quantitative analysis of length of regenerated epidermis in wounds of db/db mice in different treatment groups on day 15. Scale bars, 200 µm. n = 5 biologically independent samples. d, Representative Masson staining images in wounds of db/db mice in different treatment groups on day 15. Scale bars, 200 µm. e, Representative DHE staining images and quantitative analysis of DHE intensity in wounds of db/db mice in different treatment groups on day 15. Scale bars, 50 µm. n = 5 biologically independent samples. f, Representative α-SMA staining images and quantitative analysis of α-SMA+ vessel area in wounds of db/db mice in different treatment groups on day 15. Scale bars, 100 µm. n = 5 biologically independent samples. g, Representative CD31 staining images and quantitative analysis of capillary density in wounds of db/db mice in different treatment groups on day 15. Scale bars, 100 µm. n = 5 biologically independent samples. Control represents PBS buffer-treated group. All values are expressed as mean ± s. d. Statistical significance was determined using two-way ANOVA with Tukey’s multiple comparisons in b and two-tailed unpaired t test in c, e, f, g. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Extended Data Fig. 2 In vivo wound healing efficacy on severe diabetic mice infected by MRSA.
a, Representative images of infected diabetic wounds of severe diabetic mice in different treatment groups. Inside the green dashed line is the initial wound area. Scale bars, 2 mm. b, In vivo wound closure rates of severe diabetic mice in different treatment groups. n = 5 biologically independent samples. c, Representative DHE staining images and quantitative analysis of DHE intensity in wounds of severe diabetic mice in different treatment groups on day 15. Scale bars, 50 µm. n = 5 biologically independent samples. d, Representative H&E staining images and quantitative analysis of length of regenerated epidermis in wounds of severe diabetic mice in different treatment groups on day 15. Scale bars, 200 µm. n = 5 biologically independent samples. e, Representative α-SMA staining images and quantitative analysis of α-SMA+ vessel area in wounds of severe diabetic mice in different treatment groups. Scale bars, 100 µm. n = 5 biologically independent samples. f, Representative CD68 and CD206 staining images and quantitative analysis of the ratio of CD206+/CD68+ cells in wounds of severe diabetic mice in different treatment groups. Scale bars, 50 µm. n = 5 biologically independent samples. Control represents PBS buffer-treated group. All values are expressed as mean ± s. d. Statistical significance was determined using two-way ANOVA with Tukey’s multiple comparisons in b and two-tailed unpaired t test in c-f. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
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Wei, T., Pan, T., Peng, X. et al. Janus liposozyme for the modulation of redox and immune homeostasis in infected diabetic wounds. Nat. Nanotechnol. (2024). https://doi.org/10.1038/s41565-024-01660-y
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DOI: https://doi.org/10.1038/s41565-024-01660-y
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