Denosumab Regulates Titanium Particles-Induced Macrophages Inflammation and Osteolysis by Targeting the ST18/PARP1 Interaction

Abstract

Objective: This study aims to investigate the role of Denosumab and its downstream target ST18 in wear particle-induced macrophage inflammation and osteolysis, and to explore the underlying mechanisms involved in aseptic loosening (AL) of prosthetic joints.

Methods: Macrophages were stimulated with titanium particles (TiPs), and inflammatory responses were assessed using qRT-PCR, western blot, flow cytometry, and immunohistochemistry. Denosumab's effects on inflammation and osteolysis were evaluated with the same approaches. Potential targets of Denosumab were screened via online databases and validated by qRT-PCR and western blot. ST18 was modulated in macrophages using lentiviral overexpression and knockdown systems. A mouse calvarial model of TiPs-induced osteolysis was established, and the roles of Denosumab and ST18 were examined in vivo using micro-CT, H&E staining, and tartrate-resistant acid phosphatase (TRACP) staining.

Results: Denosumab suppressed TiPs-induced macrophage inflammation by inhibiting NF-κB signaling and M1 polarization. We identified ST18 as a direct target of Denosumab, whose expression was upregulated by TiPs but downregulated by Denosumab. Lentivirus-mediated ST18 knockdown markedly attenuated TiPs-induced inflammation, whereas ST18 overexpression exacerbated it. Further analysis revealed that ST18 interacts with PARP1. In vivo, Denosumab significantly alleviated TiPs-induced osteolysis in mouse calvaria, an effect that was reversed by ST18 overexpression.

Conclusion: Denosumab attenuates TiPs-induced macrophage activation and osteolysis through suppression of the NF-κB pathway and M1 polarization, with ST18 serving as a key mediator. These findings highlight Denosumab as a promising therapeutic candidate for the prevention of prosthesis-related aseptic loosening.

Keywords

aseptic loosening / Denosumab / macrophage inflammation / osteolysis / ST18

References

[1] J. A. Singh, S. Yu, L. Chen, and J. D. Cleveland, “Rates of Total Joint Replacement in the United States: Future Projections to 2020-2040 Using the National Inpatient Sample,” Journal of Rheumatology 46, no. 9 (2019): 1134–1140.

[2] P. Cnudde, E. Bulow, S. Nemes, Y. Tyson, M. Mohaddes, and O. Rolfson, “Association Between Patient Survival Following Reoperation After Total Hip Replacement and the Reason for Reoperation: An Analysis of 9,926 Patients in the Swedish Hip Arthroplasty Register,” Acta Orthopaedica 90, no. 3 (2019): 226–230.

[3] A. Hart, V. Janz, R. T. Trousdale, R. J. Sierra, D. J. Berry, and M. P. Abdel, “Long-Term Survivorship of Total Hip Arthroplasty With Highly Cross-Linked Polyethylene for Osteonecrosis,” Journal of Bone and Joint Surgery. American Volume 101, no. 17 (2019): 1563–1568.

[4] P. Cnudde, E. Bulow, S. Nemes, Y. Tyson, M. Mohaddes, and O. Rolfson, “Association Between Patient Survival Following Reoperation After Total Hip Replacement and the Reason for Reoperation: An Analysis of 9,926 Patients in the Swedish Hip Arthroplasty Register,” Acta Orthopaedica 90 (2019): 226–230.

[5] D. M. Vasconcelos, M. Ribeiro-da-Silva, A. Mateus, et al., “Immune Response and Innervation Signatures in Aseptic Hip Implant Loosening,” Journal of Translational Medicine 14, no. 1 (2016): 205.

[6] A. J. Rao, E. Gibon, T. Ma, Z. Yao, R. L. Smith, and S. B. Goodman, “Revision Joint Replacement, Wear Particles, and Macrophage Polarization,” Acta Biomaterialia 8, no. 7 (2012): 2815–2823.

[7] E. D. Deeks, “Denosumab: A Review in Postmenopausal Osteoporosis,” Drugs and Aging 35, no. 2 (2018): 163–173.

[8] S. Nagoya, K. Tateda, S. Okazaki, I. Kosukegawa, J. Shimizu, and T. Yamashita, “Restoration of Proximal Periprosthetic Bone Loss by Denosumab in Cementless Total Hip Arthroplasty,” European Journal of Orthopaedic Surgery and Traumatology 28, no. 8 (2018): 1601–1607.

[9] A. Nystrom, D. Kiritopoulos, G. Ullmark, et al., “Denosumab Prevents Early Periprosthetic Bone Loss After Uncemented Total Hip Arthroplasty: Results From a Randomized Placebo-Controlled Clinical Trial,” Journal of Bone and Mineral Research 35, no. 2 (2020): 239–247.

[10] M. M. Weivoda, C. K. Chew, D. G. Monroe, et al., “Identification of Osteoclast-Osteoblast Coupling Factors in Humans Reveals Links Between Bone and Energy Metabolism,” Nature Communications 11, no. 1 (2020): 87.

[11] J. Lu, D. Hu, Y. Zhang, C. Ma, L. Shen, and B. Shuai, “Current Comprehensive Understanding of Denosumab (The RANKL Neutralizing Antibody) in the Treatment of Bone Metastasis of Malignant Tumors, Including Pharmacological Mechanism and Clinical Trials,” Frontiers in Oncology 13 (2023): 1133828.

[12] E. Palmerini, E. L. Staals, L. B. Jones, D. M. Donati, A. Longhi, and R. L. Randall, “Role of (Neo)adjuvant Denosumab for Giant Cell Tumor of Bone,” Current Treatment Options in Oncology 21, no. 8 (2020): 68.

[13] H. G. Bone, R. B. Wagman, M. L. Brandi, et al., “10 Years of Denosumab Treatment in Postmenopausal Women With Osteoporosis: Results From the Phase 3 Randomised FREEDOM Trial and Open-Label Extension,” Lancet Diabetes and Endocrinology 5, no. 7 (2017): 513–523.

[14] J. Xu, H. Li, Y. Qu, et al., “Denosumab Might Prevent Periprosthetic Bone Loss After Total Hip and Knee Arthroplasties: A Review,” Art 3, no. 1 (2021): 13.

[15] D. Steinbach, P. Bader, A. Willasch, et al., “Prospective Validation of a New Method of Monitoring Minimal Residual Disease in Childhood Acute Myelogenous Leukemia,” Clinical Cancer Research 21, no. 6 (2015): 1353–1359.

[16] R. Shukla, K. R. Upton, M. Munoz-Lopez, et al., “Endogenous Retrotransposition Activates Oncogenic Pathways in Hepatocellular Carcinoma,” Cell 153, no. 1 (2013): 101–111.

[17] B. Jandrig, S. Seitz, B. Hinzmann, et al., “ST18 Is a Breast Cancer Tumor Suppressor Gene at Human Chromosome 8q11.2,” Oncogene 23, no. 57 (2004): 9295–9302.

[18] J. Yang, M. F. Siqueira, Y. Behl, M. Alikhani, and D. T. Graves, “The Transcription Factor ST18 Regulates Proapoptotic and Proinflammatory Gene Expression in Fibroblasts,” FASEB Journal 22, no. 11 (2008): 3956–3967.

[19] P. O. Hassa and M. O. Hottiger, “The Diverse Biological Roles of Mammalian PARPS, a Small but Powerful Family of Poly-ADP-Ribose Polymerases,” Frontiers in Bioscience 13 (2008): 3046–3082.

[20] J. P. Gagne, M. J. Hendzel, A. Droit, and G. G. Poirier, “The Expanding Role of Poly(ADP-Ribose) Metabolism: Current Challenges and New Perspectives,” Current Opinion in Cell Biology 18, no. 2 (2006): 145–151.

[21] P. Chambon, J. D. Weill, and P. Mandel, “Nicotinamide Mononucleotide Activation of New DNA-Dependent Polyadenylic Acid Synthesizing Nuclear Enzyme,” Biochemical and Biophysical Research Communications 11 (1963): 39–43.

[22] J. Diefenbach and A. Burkle, “Introduction to Poly(ADP-Ribose) Metabolism,” Cellular and Molecular Life Sciences 62, no. 7–8 (2005): 721–730.

[23] J. P. Chow, W. Y. Man, M. Mao, et al., “PARP1 Is Overexpressed in Nasopharyngeal Carcinoma and Its Inhibition Enhances Radiotherapy,” Molecular Cancer Therapeutics 12, no. 11 (2013): 2517–2528.

[24] F. Rojo, J. Garcia-Parra, S. Zazo, et al., “Nuclear PARP-1 Protein Overexpression Is Associated With Poor Overall Survival in Early Breast Cancer,” Annals of Oncology 23, no. 5 (2012): 1156–1164.

[25] H. Cheng, Z. Zhang, A. Borczuk, et al., “PARP Inhibition Selectively Increases Sensitivity to Cisplatin in ERCC1-Low Non-Small Cell Lung Cancer Cells,” Carcinogenesis 34, no. 4 (2013): 739–749.

[26] P. Pacher, J. G. Mabley, F. G. Soriano, L. Liaudet, and C. Szabo, “Activation of Poly(ADP-Ribose) Polymerase Contributes to the Endothelial Dysfunction Associated With Hypertension and Aging,” International Journal of Molecular Medicine 9, no. 6 (2002): 659–664.

[27] P. O. Hassa, S. S. Haenni, C. Buerki, et al., “Acetylation of Poly(ADP-Ribose) Polymerase-1 by p300/CREB-Binding Protein Regulates Coactivation of NF-kappaB-Dependent Transcription,” Journal of Biological Chemistry 280, no. 49 (2005): 40450–40464.

[28] A. Carrillo, Y. Monreal, P. Ramirez, et al., “Transcription Regulation of TNF-Alpha-Early Response Genes by Poly(ADP-Ribose) Polymerase-1 in Murine Heart Endothelial Cells,” Nucleic Acids Research 32, no. 2 (2004): 757–766.

[29] M. Robson, S. A. Im, E. Senkus, et al., “Olaparib for Metastatic Breast Cancer in Patients With a Germline BRCA Mutation,” New England Journal of Medicine 377, no. 6 (2017): 523–533.

[30] M. A. Ghonim, S. V. Ibba, A. F. Tarhuni, et al., “Targeting PARP-1 With Metronomic Therapy Modulates MDSC Suppressive Function and Enhances Anti-PD-1 Immunotherapy in Colon Cancer,” Journal for Immunotherapy of Cancer 9, no. 1 (2021): e001643.

[31] A. F. Farago, B. Y. Yeap, M. Stanzione, et al., “Combination Olaparib and Temozolomide in Relapsed Small-Cell Lung Cancer,” Cancer Discovery 9, no. 10 (2019): 1372–1387.

[32] S. Kobak, “Olaparib-Related Seronegative Inflammatory Arthritis: First Report in the Literature,” Scandinavian Journal of Rheumatology 50, no. 1 (2021): 74–75.

[33] H. Kang, Y. Yan, P. Jia, et al., “Desferrioxamine Reduces Ultrahigh-Molecular-Weight Polyethylene-Induced Osteolysis by Restraining Inflammatory Osteoclastogenesis via Heme Oxygenase-1,” Cell Death and Disease 7, no. 10 (2016): e2435.

[34] S. Li, C. Wu, S. Lin, et al., “HUCMSC-Derived Exosomes Suppress the Titanium Particles-Induced Osteolysis in Mice Through Inhibiting CCL2 and CCL3,” Orthopaedic Surgery 15, no. 3 (2023): 888–898.

[35] Y. Chen, Y. Wang, R. Tang, et al., “Dendritic Cells-Derived Interferon-lambda1 Ameliorated Inflammatory Bone Destruction Through Inhibiting Osteoclastogenesis,” Cell Death and Disease 11, no. 6 (2020): 414.

[36] J. Wang, Y. Xue, Y. Wang, et al., “BMP-2 Functional Polypeptides Relieve Osteolysis via Bi-Regulating Bone Formation and Resorption Coupled With Macrophage Polarization,” Npj Regenerative Medicine 8, no. 1 (2023): 6.

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2025 The Author(s). Orthopaedic Surgery published by Tianjin Hospital and John Wiley & Sons Australia, Ltd.