Surmounting Recalcitrant Airway Inflammatory Disorders With a Multi-Targeting Therapeutic Strategy by Tannic Acid-Modified CuInP₂S₆ Nanosheets

Abstract

Recalcitrant airway inflammatory diseases have drawn significant attention due to their high incidence and substantial healthcare costs. Analysis of clinical samples from patients with chronic rhinosinusitis with nasal polyps (CRSwNP) revealed the coexistence of neutrophilic and eosinophilic inflammation, which may account for the limited efficacy of traditional single-target therapeutic strategies. Moreover, cell-free DNA (cfDNA)—an emerging inflammatory mediator—has been implicated in both eosinophilic and neutrophilic responses through its role in the formation of extracellular traps. In this study, we developed tannic acid (TA)-modified CuInP₂S₆ (CIPS) nanosheets (C-TA₁; w/w = 1:1) as a multi-targeting therapeutic nanoplatform for recalcitrant airway inflammatory diseases. The C-TA₁ nanosheets demonstrated efficient cfDNA clearance via hydrogen bonding interactions, thereby inhibiting cfDNA-triggered toll-like receptor 9 (TLR9) activation and subsequent nuclear factor-κB (NF-κB) inflammatory signaling. Additionally, C-TA₁ exhibited potent antioxidant and antibacterial activities, which were ascribed to the inherent properties of the two-dimensional nanostructure and the chemical characteristics of TA, respectively. The in vivo therapeutic efficacy of C-TA₁ was evaluated in murine models with neutrophilic and eosinophilic airway inflammation, respectively. C-TA₁ markedly attenuated airway inflammation in both of these animal models by reducing reactive species, immune cell infiltration, goblet cell hyperplasia and the expression of pro‑inflammatory cytokines. Furthermore, treatment with C-TA₁ effectively modulated the dysregulated airway microbiota observed in the inflammatory state. Our findings demonstrate a multi-targeting nanoformulation designed to mitigate multiple key pathological drivers of severe airway inflammation concurrently. This engineered system presents a promising strategy for managing respiratory inflammatory disorders and also other inflammation-related diseases.

Keywords

airway inflammation / bacterial infection / cell-free DNA / CuInP₂S₆ nanosheets / reactive species / tannic acid

References

[1] C. Hopkins, “Chronic Rhinosinusitis With Nasal Polyps,” New England Journal of Medicine 381 (2019): 55–63, https://doi.org/10.1056/NEJMcp1800215.

[2] P. Venkatesan, “2023 GINA Report for Asthma,” Lancet Respiratory Medicine 11 (2023): 589, https://doi.org/10.1016/S2213-2600(23)00230-8.

[3] P. Venkatesan, “GOLD COPD Report: 2024 Update,” Lancet Respiratory Medicine 12 (2024): 15–16, https://doi.org/10.1016/S2213-2600(23)00461-7.

[4] C. Liu, L. Xi, Y. Liu, et al., “An Inhalable Hybrid Biomimetic Nanoplatform for Sequential Drug Release and Remodeling Lung Immune Homeostasis in Acute Lung Injury Treatment,” ACS Nano 17 (2023): 11626–11644, https://doi.org/10.1021/acsnano.3c02075.

[5] J. V. Fahy, “Type 2 Inflammation in Asthma — Present in Most, Absent in Many,” Nature Reviews Immunology 15 (2015): 57–65, https://doi.org/10.1038/nri3786.

[6] A. Kato, A. T. Peters, W. W. Stevens, R. P. Schleimer, B. K. Tan, and R. C. Kern, “Endotypes of Chronic Rhinosinusitis: Relationships to Disease Phenotypes, Pathogenesis, Clinical Findings, and Treatment Approaches,” Allergy 77 (2022): 812–826, https://doi.org/10.1111/all.15074.

[7] T. Delemarre, B. S. Bochner, H. U. Simon, and C. Bachert, “Rethinking Neutrophils and Eosinophils in Chronic Rhinosinusitis,” Journal of Allergy and Clinical Immunology 148 (2021): 327–335, https://doi.org/10.1016/j.jaci.2021.03.024.

[8] A. Ray and J. K. Kolls, “Neutrophilic Inflammation in Asthma and Association With Disease Severity,” Trends in Immunology 38 (2017): 942–954, https://doi.org/10.1016/j.it.2017.07.003.

[9] T. Delemarre, G. Holtappels, N. De Ruyck, et al., “Type 2 Inflammation in Chronic Rhinosinusitis Without Nasal Polyps: Another Relevant Endotype,” Journal of Allergy and Clinical Immunology 146 (2020): 337–343.e6, https://doi.org/10.1016/j.jaci.2020.04.040.

[10] X. Wang, N. Zhang, M. Bo, et al., “Diversity of TH Cytokine Profiles in Patients With Chronic Rhinosinusitis: A Multicenter Study in Europe, Asia, and Oceania,” Journal of Allergy and Clinical Immunology 138 (2016): 1344–1353, https://doi.org/10.1016/j.jaci.2016.05.041.

[11] J. Ruan, J. Zhao, X. Li, et al., “Characterizing the Neutrophilic Inflammation in Chronic Rhinosinusitis With Nasal Polyps,” Frontiers in Cell and Developmental Biology 9 (2021): 793073, https://doi.org/10.3389/fcell.2021.793073.

[12] J. Aucamp, A. J. Bronkhorst, C. P. S. Badenhorst, and P. J. Pretorius, “The Diverse Origins of Circulating Cell-Free DNA in the Human Body: A Critical Re-Evaluation of the Literature,” Biological Reviews of the Cambridge Philosophical Society 93 (2018): 1649–1683, https://doi.org/10.1111/brv.12413.

[13] D. S. C. Han and Y. M. D. Lo, “The Nexus of cfDNA and Nuclease Biology,” Trends in Genetics 37 (2021): 758–770, https://doi.org/10.1016/j.tig.2021.04.005.

[14] E. Heitzer, L. Auinger, and M. R. Speicher, “Cell-Free DNA and Apoptosis: How Dead Cells Inform About the Living,” Trends in Molecular Medicine 26 (2020): 519–528, https://doi.org/10.1016/j.molmed.2020.01.012.

[15] P. Jiang, K. C. A. Chan, and Y. M. D. Lo, “Liver-Derived Cell-Free Nucleic Acids in Plasma: Biology and Applications in Liquid Biopsies,” Journal of Hepatology 71 (2019): 409–421, https://doi.org/10.1016/j.jhep.2019.04.003.

[16] S. R. Paludan, L. S. Reinert, and V. Hornung, “DNA-Stimulated Cell Death: Implications for Host Defence, Inflammatory Diseases and Cancer,” Nature Reviews Immunology 19 (2019): 141–153, https://doi.org/10.1038/s41577-018-0117-0.

[17] S. Gravina, J. M. Sedivy, and J. Vijg, “The Dark Side of Circulating Nucleic Acids,” Aging Cell 15 (2016): 398–399, https://doi.org/10.1111/acel.12454.

[18] I. Naqvi, N. Giroux, L. Olson, et al., “DAMPs/PAMPs Induce Monocytic TLR Activation and Tolerance in COVID-19 Patients; Nucleic Acid Binding Scavengers Can Counteract Such TLR Agonists,” Biomaterials 283 (2022): 121393, https://doi.org/10.1016/j.biomaterials.2022.121393.

[19] Y. Xiao, H. Fang, Y. Zhu, et al., “Multifunctional Cationic Hyperbranched Polyaminoglycosides That Target Multiple Mediators for Severe Abdominal Trauma Management,” Advancement of Science 11 (2024): e2305273, https://doi.org/10.1002/advs.202305273.

[20] A. Marshak-Rothstein and I. R. Rifkin, “Immunologically Active Autoantigens: The Role of Toll-Like Receptors in the Development of Chronic Inflammatory Disease,” Annual Review of Immunology 25 (2007): 419–441, https://doi.org/10.1146/annurev.immunol.22.012703.104514.

[21] E. Vringer and S. W. G. Tait, “Mitochondria and Cell Death-Associated Inflammation,” Cell Death and Differentiation 30 (2023): 304–312, https://doi.org/10.1038/s41418-022-01094-w.

[22] K. Wu, X. Lu, Y. Li, et al., “Polyglycerol-Amine Covered Nanosheets Target Cell-Free DNA to Attenuate Acute Kidney Injury,” Advancement of Science 10 (2023): e2300604, https://doi.org/10.1002/advs.202300604.

[23] Z. Tu, M. Liu, C. Xu, et al., “Functional 2D Nanoplatforms Alleviate Eosinophilic Chronic Rhinosinusitis by Modulating Eosinophil Extracellular Trap Formation,” Advancement of Science 11 (2024): e2307800, https://doi.org/10.1002/advs.202307800.

[24] L. Ronchetti, I. Terrenato, M. Ferretti, et al., “Circulating Cell Free DNA and Citrullinated Histone H₃ as Useful Biomarkers of NETosis in Endometrial Cancer,” Journal of Experimental & Clinical Cancer Research 41 (2022): 151, https://doi.org/10.1186/s13046-022-02359-5.

[25] M. Qu, W. Xu, X. Zhou, et al., “An ROS-Scavenging Treg-Recruiting Hydrogel Patch for Diabetic Wound Healing,” Advanced Functional Materials 34 (2024): 2314500, https://doi.org/10.1002/adfm.202314500.

[26] J. Dan Dunn, L. A. Alvarez, X. Zhang, and T. Soldati, “Reactive Oxygen Species and Mitochondria: A Nexus of Cellular Homeostasis,” Redox Biology 6 (2015): 472–485, https://doi.org/10.1016/j.redox.2015.09.005.

[27] K. Wu, M. Fu, Y. Zhao, et al., “Anti-Oxidant Anti-Inflammatory and Antibacterial Tannin-Crosslinked Citrate-Based Mussel-Inspired Bioadhesives Facilitate Scarless Wound Healing,” Bioactive Materials 20 (2023): 93–110.

[28] X. Zhang, H. Li, Y. Liu, et al., “Acid-Responsive CST@NPs Enhanced Diabetic Wound Healing Through Rescuing Mitochondrial Dysfunction,” Bioactive Materials 44 (2025): 269–282.

[29] B. C. Dickinson and C. J. Chang, “Chemistry and Biology of Reactive Oxygen Species in Signaling or Stress Responses,” Nature Chemical Biology 7 (2011): 504–511, https://doi.org/10.1038/nchembio.607.

[30] Y. Lee, K. Sugihara, M. G. R. Gillilland, S. Jon, N. Kamada, and J. J. Moon, “Hyaluronic Acid–Bilirubin Nanomedicine for Targeted Modulation of Dysregulated Intestinal Barrier, Microbiome and Immune Responses in Colitis,” Nature Materials 19 (2020): 118–126, https://doi.org/10.1038/s41563-019-0462-9.

[31] R. Nie, J. Zhang, Q. Jia, et al., “Structurally Oriented Carbon Dots as ROS Nanomodulators for Dynamic Chronic Inflammation and Infection Elimination,” ACS Nano 18 (2024): 22055–22070, https://doi.org/10.1021/acsnano.4c05266.

[32] G. Li, H. Deng, W. Xu, et al., “Combating Antibiotic-Resistant Bacterial Infection Using Coassembled Dimeric Antimicrobial Peptide-Based Nanofibers,” ACS Nano 19 (2025): 3155–3171, https://doi.org/10.1021/acsnano.4c09347.

[33] Z. Geng, Z. Cao, and J. Liu, “Recent Advances in Targeted Antibacterial Therapy Basing on Nanomaterials,” Exploration (Beijing) 3 (2023): 20210117, https://doi.org/10.1002/EXP.20210117.

[34] R. Li, J. Li, and X. Zhou, “Lung Microbiome: New Insights Into the Pathogenesis of Respiratory Diseases,” Signal Transduction and Targeted Therapy 9 (2024): 19, https://doi.org/10.1038/s41392-023-01722-y.

[35] P. H. H. Schneeberger, J. Prescod, L. Levy, D. Hwang, T. Martinu, and B. Coburn, “Microbiota Analysis Optimization for Human Bronchoalveolar Lavage Fluid,” Microbiome 7 (2019): 141, https://doi.org/10.1186/s40168-019-0755-x.

[36] H. Dai, Q. Fan, and C. Wang, “Recent Applications of Immunomodulatory Biomaterials for Disease Immunotherapy,” Exploration (Beijing) 2 (2022): 20210157, https://doi.org/10.1002/EXP.20210157.

[37] K. Parvez and C. Casiraghi, “Biocompatible 2D Material Inks Enabled by Supramolecular Chemistry: From Synthesis to Applications,” Accounts of Chemical Research 58 (2025): 189–198, https://doi.org/10.1021/acs.accounts.4c00596.

[38] M. Park, J. Y. Yang, M. J. Yeom, et al., “An Artificial Neuromuscular Junction for Enhanced Reflexes and Oculomotor Dynamics Based on a Ferroelectric CuInP₂S₆/GaN HEMT,” Science Advances 9 (2023): eadh9889, https://doi.org/10.1126/sciadv.adh9889.

[39] G. Zhang, Y. Cong, F. Liu, et al., “A Nanomaterial Targeting the Spike Protein Captures SARS-CoV-2 Variants and Promotes Viral Elimination,” Nature Nanotechnology 17 (2022): 993–1003, https://doi.org/10.1038/s41565-022-01177-2.

[40] D. Mu, W. Wang, J. Li, et al., “Ultrasmall Fe(III)-Tannic Acid Nanoparticles to Prevent Progression of Atherosclerotic Plaques,” ACS Applied Materials & Interfaces 13 (2021): 33915–33925, https://doi.org/10.1021/acsami.1c09480.

[41] T. Liu, M. Zhang, W. Liu, et al., “Metal Ion/Tannic Acid Assembly as a Versatile Photothermal Platform in Engineering Multimodal Nanotheranostics for Advanced Applications,” ACS Nano 12 (2018): 3917–3927, https://doi.org/10.1021/acsnano.8b01456.

[42] L. Xie, J. Li, L. Wang, and Y. Dai, “Engineering Metal-Phenolic Networks for Enhancing Cancer Therapy by Tumor Microenvironment Modulation,” Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology 15 (2023): e1864, https://doi.org/10.1002/wnan.1864.

[43] S. Tao, Y. Yang, C. Wu, et al., “Nanocapsuled Neutrophil Extracellular Trap Scavenger Combating Chronic Infectious Bone Destruction Diseases,” Advancement of Science 12 (2025): e2411274, https://doi.org/10.1002/advs.202411274.

[44] S. Wang, Q. Guo, R. Xu, P. Lin, G. Deng, and X. Xia, “Combination of Ferroptosis and Pyroptosis Dual Induction by Triptolide Nano-MOFs for Immunotherapy of Melanoma,” Journal of Nanobiotechnology 21 (2023): 383, https://doi.org/10.1186/s12951-023-02146-0.

[45] Q. Zhang, M. Raoof, Y. Chen, et al., “Circulating Mitochondrial DAMPs Cause Inflammatory Responses to Injury,” Nature 464 (2010): 104–107, https://doi.org/10.1038/nature08780.

[46] A. J. van der Meer, A. Kroeze, A. J. Hoogendijk, et al., “Systemic Inflammation Induces Release of Cell-Free DNA From Hematopoietic and Parenchymal Cells in Mice and Humans,” Blood Advances 3 (2019): 724–728, https://doi.org/10.1182/bloodadvances.2018018895.

[47] L. Tumburu, S. Ghosh-Choudhary, F. T. Seifuddin, et al., “Circulating Mitochondrial DNA is a Proinflammatory DAMP in Sickle Cell Disease,” Blood 137 (2021): 3116–3126, https://doi.org/10.1182/blood.2020009063.

[48] M. Shin, J. H. Ryu, J. P. Park, K. Kim, J. W. Yang, and H. Lee, “DNA/Tannic Acid Hybrid Gel Exhibiting Biodegradability, Extensibility, Tissue Adhesiveness, and Hemostatic Ability,” Advanced Functional Materials 25 (2015): 1270–1278, https://doi.org/10.1002/adfm.201403992.

[49] S. Z. Alasmari, “In Vivo Imaging of Neutrophil Extracellular Traps (NETs): Visualization Methods and Outcomes,” BioMed Research International 2020 (2020): 4192745, https://doi.org/10.1155/2020/4192745.

[50] T. Li, Z. Zhang, X. Li, et al., “Neutrophil Extracellular Traps: Signaling Properties and Disease Relevance,” Mediators of Inflammation 2020 (2020): 9254087.

[51] V. Brinkmann, U. Reichard, C. Goosmann, et al., “Neutrophil Extracellular Traps Kill Bacteria,” Science 303 (2004): 1532–1535, https://doi.org/10.1126/science.1092385.

[52] O. Takeuchi and S. Akira, “Pattern Recognition Receptors and Inflammation,” Cell 140 (2010): 805–820, https://doi.org/10.1016/j.cell.2010.01.022.

[53] M. Zhong, P. Liang, Z. Feng, et al., “A Nanocomposite Competent to Overcome Cascade Drug Resistance in Ovarian Cancer via Mitochondria Dysfunction and NO Gas Synergistic Therapy,” Asian Journal of Pharmaceutical Sciences 18 (2023): 100872.

[54] Y. Yu, J. Wu, C. Lin, et al., “Elimination of Methicillin-Resistant Staphylococcus aureus Biofilms on Titanium Implants via Photothermally-Triggered Nitric Oxide and Immunotherapy for Enhanced Osseointegration,” Military Medical Research 10 (2023): 21.

[55] Z. Xuan, Z. Yun-Fei, J. Qi-Lin, and Z. Fei-Ran, “Research Progress on Antibacterial of Novel Two-Dimensional Layered Nanomaterials,” Journal of Materials Engineering 49, no. 1 (2021): 55–64.

[56] Z. Tu, Y. Zhu, W. Gao, et al., “Tackling Severe Neutrophilic Inflammation in Airway Disorders With Functionalized Nanosheets,” ACS Nano 18 (2024): 7084–7097, https://doi.org/10.1021/acsnano.3c11139.

[57] K. Tarasova, M. B. Arteaga, A. Kidtiwong, et al., “Dexamethasone: A Double-Edged Sword in the Treatment of Osteoarthritis,” Scientific Reports 15 (2025): 11832, https://doi.org/10.1038/s41598-025-96050-2.

[58] J. A. W. Polderman, V. Farhang-Razi, S. Van Dieren, et al., “Adverse Side-Effects of Dexamethasone in Surgical Patients—An Abridged Cochrane Systematic Review,” Anaesthesia 74 (2019): 929–939, https://doi.org/10.1111/anae.14610.

[59] L. N. Segal, J. C. Clemente, J. J. Tsay, et al., “Enrichment of the Lung Microbiome With Oral Taxa is Associated With Lung Inflammation of a Th17 Phenotype,” Nature Microbiology 1 (2016): 16031, https://doi.org/10.1038/nmicrobiol.2016.31.

[60] K. Yadava, C. Pattaroni, A. K. Sichelstiel, et al., “Microbiota Promotes Chronic Pulmonary Inflammation by Enhancing IL-17A and Autoantibodies,” American Journal of Respiratory and Critical Care Medicine 193 (2016): 975–987, https://doi.org/10.1164/rccm.201504-0779OC.

[61] F. Zhao, M. Hao, Q. Yue, et al., “Community Diversity and Composition Affect Ecosystem Multifunctionality Across Environmental Gradients in Boreal and Temperate Forests,” Ecological Indicators 159 (2024): 111692, https://doi.org/10.1016/j.ecolind.2024.111692.

[62] S. Mooyottu, M. S. Muyyarikkandy, F. Yousefi, et al., “Fecal Microbiota Transplantation Modulates Jejunal Host-Microbiota Interface in Weanling Piglets,” Microbiome 13 (2025): 45, https://doi.org/10.1186/s40168-025-02042-9.

[63] X. Zhang, H. Yang, Y. He, et al., “Yeast-Inspired Orally-Administered Nanocomposite Scavenges Oxidative Stress and Restores Gut Immune Homeostasis for Inflammatory Bowel Disease Treatment,” ACS Nano 19, no. 7 (2025): 7350–7369.

[64] H. Li, M. Liu, S. Zhang, et al., “Construction of CpG Delivery Nanoplatforms by Functionalized MoS₂ Nanosheets for Boosting Antitumor Immunity in Head and Neck Squamous Cell Carcinoma,” Small 19 (2023): e2300380, https://doi.org/10.1002/smll.202300380.

[65] Y. Deng, Z. Zhao, M. Sheldon, et al., “LIFR Regulates Cholesterol-Driven Bidirectional Hepatocyte–Neutrophil Cross-Talk to Promote Liver Regeneration,” Nature Metabolism 6 (2024): 1756–1774, https://doi.org/10.1038/s42255-024-01110-y.

[66] J. Wang, C. Deng, M. Liu, et al., “A Fourth Dose of the Inactivated SARS-CoV-2 Vaccine Redistributes Humoral Immunity to the N-Terminal Domain,” Nature Communications 13 (2022): 6866, https://doi.org/10.1038/s41467-022-34633-7.

[67] L. S. van Rijt, S. Jung, A. Kleinjan, et al., “In Vivo Depletion of Lung CD11c+ Dendritic Cells During Allergen Challenge Abrogates the Characteristic Features of Asthma,” Journal of Experimental Medicine 201 (2005): 981–991, https://doi.org/10.1084/jem.20042311.

[68] H. Harb, E. Stephen-Victor, E. Crestani, et al., “A Regulatory T Cell Notch4–GDF15 Axis Licenses Tissue Inflammation in Asthma,” Nature Immunology 21 (2020): 1359–1370, https://doi.org/10.1038/s41590-020-0777-3.

RIGHTS & PERMISSIONS

2026 The Author(s). Exploration published by Henan University and John Wiley & Sons Australia, Ltd.