Abstract
Joint contracture resulting from prolonged immobilization is a common complication characterized by pain, stiffness, and limited mobility. Although inflammation and tissue remodeling are known contributors, the underlying cellular mechanisms remain unclear. Here, we investigated the involvement of Caspase-1/GSDMD-mediated pyroptosis in immobilization-induced knee joint contracture and evaluated the therapeutic potential of disulfiram (DSF), an FDA-approved drug repurposed here based on its reported anti-pyroptotic activity. In a rat model of knee immobilization, we assessed joint mobility, histopathological changes, and molecular markers associated with pyroptosis. Immobilization significantly induced contracture, inflammatory infiltration, and activation of the Caspase-1/GSDMD pathway in the joint capsule. Treatment with DSF improved joint range of motion, reduced tissue damage, and reduced caspase-1/GSDMD-related pyroptosis markers and GSDMD cleavage. Notably, DSF decreased IL-1β and IL-18 protein levels without affecting their mRNA expression. Together, these findings support a role for Caspase-1/GSDMD-associated pyroptosis in immobilization-induced joint contracture and suggest that DSF alleviates contracture, at least in part, by modulating this pathway, providing new mechanistic insight and a potential therapeutic approach for preventing post-immobilization joint contracture.
Data availability
Data will be made available on reasonable request.
References
Zhou, Y. et al. Rabbit model of extending knee joint contracture: Progression of joint motion restriction and subsequent joint capsule changes after immobilization. J. Knee Surg. 33, 15–21. https://doi.org/10.1055/s-0038-1676502 (2020).
Kaneguchi, A. & Ozawa, J. Inflammation and fibrosis induced by joint remobilization, and relevance to progression of arthrogenic joint contracture: A narrative review. Physiol. Res. 71, 447–488. https://doi.org/10.33549/physiolres.934876 (2022).
Xiong, B. et al. Distribution pattern of dynamic plantar pressure in patients with traumatic extension knee contracture. Chin. Gen. Pract. https://doi.org/10.12114/j.issn.1007-9572.2022.0546 (2022).
Wang, F. et al. Effect of radial extracorporeal shock wave combined with conventional rehabilitation on post-traumatic knee extension contracture. Chin. J. Bone Joint Injury. 35 (3). CNKI:SUN:GGJS.0.2020-02-029 (2020).
Xu, Q., Zhang, Q., Zhou, Y. & Wang, F. Short-term efficacy of bedside static progressive stretch therapy for knee extension contracture. Chin. J. Trauma. Disabil. Med. 29, 10–11. https://doi.org/10.13214/j.cnki.cjotadm.2021.16.004 (2021).
Zhang, Q. B., Zhang, R., Zhou, Y., Wang, Q. N. & Wang, H. Risk factors analysis of joint contracture in patients with injuries of knee and surrounding tissues. Chin. J. Clin. Healthc. 27, 558–561. https://doi.org/10.3969/J.issn.1672-6790.2024.04.029 (2024).
Mao, J. et al. Extracorporeal shock wave and melatonin alleviate joint capsule fibrosis after knee trauma in rats by regulating autophagy. Curr. Mol. Med. https://doi.org/10.2174/0115665240339436240909100847 (2024).
Li, X. M., Wang, K., Liu, M., Zhang, Q. B. & Zhou, Y. Nicorandil mitigates arthrogenic contracture induced by knee joint extension immobilization in rats: Interference with RhoA/ROCK signaling and TGF-β1/Smad pathway. Eur. Cells Mater. 47, 59–72. https://doi.org/10.22203/eCM.v047a05 (2024).
Huang, P. P. et al. Effect of radial extracorporeal shock wave combined with ultrashort wave diathermy on fibrosis and contracture of muscle. Am. J. Phys. Med. Rehabil. 100, 643–650. https://doi.org/10.1097/PHM.0000000000001599 (2021).
Liu, Z. et al. Emergent peptides of the antifibrotic arsenal: Taking aim at myofibroblast promoting pathways. Biomolecules. https://doi.org/10.3390/biom13081179 (2023).
Hu, B., Phan, S. & Myofibroblasts Curr. Opin. Rheumatol. 25, 71–77. https://doi.org/10.1097/BOR.0b013e32835b1352 (2013).
Rao, Z. et al. Pyroptosis in inflammatory diseases and cancer. Theranostics 12, 4310–4329. https://doi.org/10.7150/thno.71086 (2022).
Guan, M. et al. Mechanisms of chondrocyte cell death in osteoarthritis: implications for disease progression and treatment. J. Orthop. Surg. Res. 19, 550. https://doi.org/10.1186/s13018-024-05055-6 (2024).
Kayagaki, N. et al. Caspase-11 cleaves gasdermin D for non-canonical inflammasome signalling. Nature 526, 666–671 (2015).
Shi, J. et al. Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death. Nature 526, 660–665 (2015).
Chang, X. et al. Pyroptosis: A novel intervention target in the progression of osteoarthritis. J. Inflamm. Res. 15, 3859–3871. https://doi.org/10.2147/JIR.S368501 (2022).
Pan, Y. et al. Pyroptosis in development, inflammation and disease. Front. Immunol. 13, 991044. https://doi.org/10.3389/fimmu.2022.991044 (2022).
Savin, I. A., Zenkova, M. A. & Sen’kova, A. V. Pulmonary fibrosis as a result of acute lung inflammation: Molecular mechanisms, relevant in vivo models, prognostic and therapeutic approaches. Int. J. Mol. Sci. https://doi.org/10.3390/ijms232314959 (2022).
Wu, Q., Du, J., Bae, E. J. & Choi, Y. Pyroptosis in skeleton diseases: A potential therapeutic target based on inflammatory cell death. Int. J. Mol. Sci. https://doi.org/10.3390/ijms25169068 (2024).
Rozi, R., Zhou, Y., Rong, K. & Chen, P. miR-124-3p sabotages lncRNA MALAT1 stability to repress chondrocyte pyroptosis and relieve cartilage injury in osteoarthritis. J. Orthop. Surg. Res. 17, 453. https://doi.org/10.1186/s13018-022-03334-8 (2022).
Gong, Y. et al. Maltol ameliorates intervertebral disc degeneration through inhibiting PI3K/AKT/NF-kappaB pathway and regulating NLRP3 inflammasome-mediated pyroptosis. Inflammopharmacology 31, 369–384. https://doi.org/10.1007/s10787-022-01098-5 (2023).
Yang, X. M. et al. Disulfiram inhibits liver fibrosis in rats by suppressing hepatic stellate cell activation and viability. BMC Pharmacol. Toxicol. https://doi.org/10.1186/s40360-022-00583-5 (2022).
Bernier, M. et al. Disulfiram treatment normalizes body weight in obese mice. Cell Metabol. 32, 203–214. https://doi.org/10.1016/j.cmet.2020.04.019 (2020).
Traughber, C. A. et al. Disulfiram reduces atherosclerosis and enhances efferocytosis, autophagy, and atheroprotective gut microbiota in hyperlipidemic mice. J. Am. Heart Assoc. 13, e033881 (2024).
Reinhardt, S. et al. Identification of disulfiram as a secretase-modulating compound with beneficial effects on Alzheimer’s disease hallmarks. Sci. Rep. 8, 1329 (2018).
Du Sert, N. P. et al. The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. PLoS Biol. 18, e3000410 (2020).
Council, N. R. et al. Guide for the care and use of laboratory animals. (2010).
Yuan, H., Wang, K., Zhang, Q. B., Wang, F. & Zhou, Y. The effect of extracorporeal shock wave on joint capsule fibrosis based on A2AR-Nrf2/HO-1 pathway in a rat extending knee immobilization model. J. Orthop. Surg, Res. 18, 930 (2023).
Zhang, R., Zhang, Q. B., Zhou, Y., Zhang, R. & Wang, F. Possible mechanism of static progressive stretching combined with extracorporeal shock wave therapy in reducing knee joint contracture in rats based on MAPK/ERK pathway. Biomolecules Biomed. 23, 277. https://doi.org/10.17305/bjbms.2022.8152 (2023).
Zhou, C. X., Wang, F., Zhou, Y., Fang, Q. Z. & Zhang, Q. B. Formation process of extension knee joint contracture following external immobilization in rats. World J. Orthop. 14, 669–681. https://doi.org/10.5312/wjo.v14.i9.669 (2023).
Abdel, M. P. et al. The Chitranjan S. Ranawat Award: Manipulation under anesthesia to treat postoperative stiffness after total knee arthroplasty: A multicenter randomized clinical trial. J. Arthroplast. 39, S9–S14 (2024).
Song, Z., Gong, Q., Guo, J. & Pyroptosis mechanisms and links with fibrosis. Cells. https://doi.org/10.3390/cells10123509 (2021).
Alyaseer, A. A. A., de Lima, M. H. S. & Braga, T. T. The role of NLRP3 inflammasome activation in the epithelial to mesenchymal transition process during the fibrosis. Front. Immunol. 11, 883. https://doi.org/10.3389/fimmu.2020.00883 (2020).
Hu, J. J. et al. FDA-approved disulfiram inhibits pyroptosis by blocking gasdermin D pore formation. Nat. Immunol. 21, 736–745. https://doi.org/10.1038/s41590-020-0669-6 (2020).
Hinz, B. & Lagares, D. Evasion of apoptosis by myofibroblasts: A hallmark of fibrotic diseases. Nat. Rev. Rheumatol. 16, 11–31. https://doi.org/10.1038/s41584-019-0324-5 (2020).
Schwarz, U. S. & Gardel, M. L. United we stand: Integrating the actin cytoskeleton and cell-matrix adhesions in cellular mechanotransduction. J. Cell. Sci. 125, 3051–3060. https://doi.org/10.1242/jcs.093716 (2012).
Tian, G. & Ren, T. Mechanical stress regulates the mechanotransduction and metabolism of cardiac fibroblasts in fibrotic cardiac diseases. Eur. J. Cell. Biol. 102, 151288. https://doi.org/10.1016/j.ejcb.2023.151288 (2023).
Lagares, D. et al. Targeted apoptosis of myofibroblasts with the BH3 mimetic ABT-263 reverses established fibrosis. Sci. Transl. Med. https://doi.org/10.1126/scitranslmed.aal3765 (2017).
Liu, F. et al. Feedback amplification of fibrosis through matrix stiffening and COX-2 suppression. J. Cell. Biol. 190, 693–706. https://doi.org/10.1083/jcb.201004082 (2010).
Ran, L. et al. KCNN4 links PIEZO-dependent mechanotransduction to NLRP3 inflammasome activation. Sci. Immunol. 8, eadf4699. https://doi.org/10.1126/sciimmunol.adf4699 (2023).
Che, Y. et al. GSDMD-dependent neutrophil extracellular traps mediate portal vein thrombosis and associated fibrosis in cirrhosis. Int. J. Mol. Sci.. https://doi.org/10.3390/ijms25169099 (2024).
Zhang, W. J., Chen, S. J., Zhou, S. C., Wu, S. Z. & Wang, H. Inflammasomes and fibrosis. Front. Immunol. 12, 643149. https://doi.org/10.3389/fimmu.2021.643149 (2021).
Udomsinprasert, W. Interleukin-1 family cytokines in liver cell death: A new therapeutic target for liver diseases. Expert Opin. Ther. Targets. 27, 1125–1143. https://doi.org/10.1080/14728222.2023.2285763 (2023).
Yunna, C., Mengru, H., Lei, W. & Weidong, C. Macrophage M1/M2 polarization. Eur. J. Pharmacol. 877, 173090. https://doi.org/10.1016/j.ejphar.2020.173090 (2020).
Nozaki, Y. et al. Inhibition of the IL-18 receptor signaling pathway ameliorates disease in a murine model of rheumatoid arthritis. Cells. https://doi.org/10.3390/cells9010011 (2019).
Ihim, S. A. et al. Interleukin-18 cytokine in immunity, inflammation, and autoimmunity: Biological role in induction, regulation, and treatment. Front. Immunol. 13, 919973. https://doi.org/10.3389/fimmu.2022.919973 (2022).
Hirooka, Y. & Nozaki, Y. Interleukin-18 in inflammatory kidney disease. Front. Med. (Lausanne). 8, 639103. https://doi.org/10.3389/fmed.2021.639103 (2021).
Matsui, F., Rhee, A., Hile, K. L., Zhang, H. & Meldrum, K. K. IL-18 induces profibrotic renal tubular cell injury via STAT3 activation. Am. J. Physiol. Ren. Physiol. 305, F1014–1021. https://doi.org/10.1152/ajprenal.00620.2012 (2013).
Knorr, J. et al. Interleukin-18 signaling promotes activation of hepatic stellate cells in mouse liver fibrosis. Hepatology 77, 1968–1982. https://doi.org/10.1002/hep.32776 (2023).
He, W. T. et al. Gasdermin D is an executor of pyroptosis and required for interleukin-1beta secretion. Cell. Res. 25, 1285–1298. https://doi.org/10.1038/cr.2015.139 (2015).
Zhao, J. et al. Disulfiram alleviates acute lung injury and related intestinal mucosal barrier impairment by targeting GSDMD-dependent pyroptosis. J. Inflamm. (Lond). 19, 17. https://doi.org/10.1186/s12950-022-00313-y (2022).
Zhou, Q. et al. Disulfiram suppressed peritendinous fibrosis through inhibiting macrophage accumulation and its pro-inflammatory properties in tendon bone healing. Front. Bioeng. Biotechnol. 10, 823933. https://doi.org/10.3389/fbioe.2022.823933 (2022).
Bai, Y. et al. Disulfiram blocks inflammatory TLR4 signaling by targeting MD-2. Proc. Natl. Acad. Sci. 120, e2306399120 (2023).
Funding
This work was supported by Health Research Program of Anhui (AHWJ2022B063); National Natural Science Incubation Program of the Second Hospital of Anhui Medical University (2022GMFY05); Summit Discipline Construction Project of Anhui Medical University (Clinical Medicine) in 2022 (2022GFXK-EFY08); Clinical Medicine Discipline Construction Project of Anhui Medical University in 2023 (2023lcxkEFY010); Health Research Program of Anhui (AHWJ2023A30077); Anhui Provincial Natural Science Foundation (2408085QH270);Key Natural Science Research Project of Anhui Educational Committee(No. 2024AH050788); Cultivation Project of Anhui Institute of Translational Medicine(2023zhyx-C85).
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Qian Lu : Writing–original draft, Project Design, Investigation, Data Curation. Xiuli Kan : Writing–review & editing, Methodology, Investigation, Data Curation. Quanbing Zhang : Methodology, Investigation, Project administration. Deting Zhu : Project administration, Methodology. Zunyu Du : Methodology, Data analysis. Xueming Li : Methodology, Investigation. Han Xiao : Project administration, Methodology. Jing Mao : Project administration, Methodology. Run Zhang : Writing–review & editing, Methodology. Yun Zhou: Writing–review & editing, Funding acquisition, Conceptualization, Supervision. All authors read and approved the final manuscript.
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Lu, Q., Kan, X., Zhang, Q. et al. Disulfiram attenuates immobilization-induced knee joint contracture by suppressing Caspase-1/GSDMD-mediated pyroptosis. Sci Rep (2026). https://doi.org/10.1038/s41598-026-42560-6
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DOI: https://doi.org/10.1038/s41598-026-42560-6
