journal article·2026-03-17Open access

Cytocompatibility Assessment of L-PBF-Manufactured Zinc–Silver–Copper Alloys for Customized Biodegradable Medical Implants

Barbara Illing, Jacob Schultheiss, Lukas Schumacher, Evi Kimmerle-Mueller, Ariadne Roehler, Alexander Heiss, Ulrich E. Klotz, Victor O. Okafor, Stefanie Krajewski, Frank Rupp

Abstract

Biodegradable zinc (Zn) has attracted increasing interest as a material for temporary implants, primarily due to its moderate degradation kinetics. In recent years, additive manufacturing of Zn alloys using the laser powder bed fusion method (L-PBF) has shown promising results. Compared to as-cast Zn alloys, it offers preferable customized solutions for patient-specific temporary biomedical implants. Due to the novelty of these printed degradable biomaterials and due to reported cytotoxic effects of Zn alloys, this study investigates additively manufactured ZnAgCu, ZnAgCuMn, and ZnAgCuTi alloys, both in as-printed and post-processed conditions, with a focus on L929 and SAOS-2 biocompatibility. In this work, we demonstrate that the increased porosity and therefore larger surface areas compared to polished Zn-alloy samples affect their biocompatibility. Minimal to no cell proliferation was observed on and near the Zn-alloy test plates after 24 h. Undiluted extracts from as-cast Zn and L-PBF-manufactured plates were initially cytotoxic to SAOS-2 cells. However, as passivation proceeded, cytocompatibility was significantly increased from day 3 onward. Zn2+ ion release peaked at 24 h and declined significantly from day 2 to day 10. Compared to the other Zn alloys, ZnAgCuMn exhibited the lowest cytocompatibility. Most intriguingly, 3-month surfaces exhibited reduced cytocompatibility to osteoblasts compared to freshly polished samples. The observed in vitro cytotoxicity motivates further investigation of as-printed and post-processed L-PBF-manufactured Zn alloys, aiming to develop novel surface modification strategies to mitigate the initial ion burst responsible for reduced cytocompatibility and to adjust and tailor the overall degradation kinetics to physiologically tolerable levels tailored to the intended clinical application.
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Research context

OpenAlex topics: Additive Manufacturing Materials and Processes · Bone Tissue Engineering Materials · Magnesium Alloys: Properties and Applications

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Affiliations

Department of Medical Materials Science and Technology, Institute of Biomedical Engineering, University Hospital Tübingen, Osianderstrasse 2-8, 72076 Tübingen, Germany

Department of Physical Metallurgy, fem Research Institute, Katharinenstraße 17, 73525 Schwäbisch Gmünd, Germany

Funding

Federal Ministry for Economic Affairs and Climate Action · Open Access Publishing Fund of the University of Tübingen

References (95)

Show source-provided references
1. Loffredo, S., Paternoster, C., and Mantovani, D. (2019). Iron-Based Degradable Implants. Encycl. Biomed. Eng., 374–385.10.1016/B978-0-12-801238-3.11023-2 ↗
2. Development of a novel biodegradable porous iron-based implant for bone replacement10.1038/s41598-020-66289-y ↗
3. Progress in manufacturing and processing of degradable Fe-based implants: A review10.1007/s40204-022-00189-4 ↗
4. Biodegradable Mg alloys for orthopedic implants—A review10.1016/j.jma.2021.06.024 ↗
5. Biodegradable Magnesium-Based Implants in Orthopedics-A General Review and Perspectives10.1002/advs.201902443 ↗
6. Thomas, K.K., Zafar, M.N., Pitt, W.G., and Husseini, G.A. (2023). Biodegradable Magnesium Alloys for Biomedical Implants: Properties, Challenges, and Surface Modifications with a Focus on Orthopedic Fixation Repair. Appl. Sci., 14.10.20944/preprints202310.1485.v1 ↗
7. A review of current challenges and prospects of magnesium and its alloy for bone implant applications
8. Long-term biocompatibility of a corrodible peripheral iron stent in the porcine descending aorta10.1016/j.biomaterials.2006.05.029 ↗
9. Dilemmas and countermeasures of Fe-based biomaterials for next-generation bone implants10.1016/j.jmrt.2022.07.089 ↗
10. Prasadh, S., Raguraman, S., Wong, R., and Gupta, M. (2022). Current Status and Outlook of Temporary Implants (Magnesium/Zinc) in Cardiovascular Applications. Metals, 12.10.3390/met12060999 ↗
11. Biodegradable magnesium alloys for orthopaedic applications: A review on corrosion, biocompatibility and surface modifications10.1016/j.msec.2016.06.020 ↗
12. U.S. Department of Health and Human Services, AT.S.D.R (2005). Toxicological Profile for Zinc, U.S. Department of Health and Human Services.
13. Zinc deficiency suppresses matrix mineralization and retards osteogenesis transiently with catch-up possibly through Runx 2 modulation10.1016/j.bone.2009.11.003 ↗
14. Role of zinc in bone formation and bone resorption10.1002/(SICI)1520-670X(1998)11:2/3<119::AID-JTRA5>3.0.CO;2-3 ↗
15. The Role of Zinc in Bone Tissue Health and Regeneration-a Review10.1007/s12011-023-03631-1 ↗
16. Feasibility, challenges and future prospects of biodegradable zinc alloys as orthopedic internal fixation implants
17. Biodegradable Metals for Cardiovascular Stents: From Clinical Concerns to Recent Zn-Alloys10.1002/adhm.201501019 ↗
18. Structural and temporal dynamics analysis of zinc-based biomaterials: History, research hotspots and emerging trends
19. Kong, L., Heydari, Z., Lami, G.H., Saberi, A., Baltatu, M.S., and Vizureanu, P. (2023). A Comprehensive Review of the Current Research Status of Biodegradable Zinc Alloys and Composites for Biomedical Applications. Materials, 16.10.3390/ma16134797 ↗
20. Long-term surveillance of zinc implant in murine artery: Surprisingly steady biocorrosion rate10.1016/j.actbio.2017.05.045 ↗
21. Challenges in the use of zinc and its alloys as biodegradable metals: Perspective from biomechanical compatibility10.1016/j.actbio.2019.07.038 ↗
22. Heiss, A., Thatikonda, V.S., Richter, A., Schmitt, L.Y., Park, D., and Klotz, U.E. (2023). Development, Processing and Aging of Novel Zn-Ag-Cu Based Biodegradable Alloys. Materials, 16.10.3390/ma16083198 ↗
23. Li, P., Schille, C., Schweizer, E., Kimmerle-Muller, E., Rupp, F., Han, X., Heiss, A., Richter, A., Legner, C., and Klotz, U.E. (2019). Evaluation of a Zn-2Ag-1.8Au-0.2V Alloy for Absorbable Biocompatible Materials. Materials, 13.10.3390/ma13010056 ↗
24. Biodegradable Zn-1.5Cu-1.5Ag alloy with anti-aging ability and strain hardening behavior for cardiovascular stents10.1016/j.msec.2020.111172 ↗
25. Fabrication, mechanical properties and in vitro degradation behavior of newly developed ZnAg alloys for degradable implant applications10.1016/j.msec.2017.04.023 ↗
26. 380 MPa-30% grade biodegradable Zn-Mn-Mg-Ca alloy: Bimodal grain structure, large work-hardening strain, and enhanced biocompatibility10.1016/j.actbio.2024.12.050 ↗
27. Magnesium-induced strengthening, degradation and osteogenesis for additively manufactured Zn-Mg orthopedic implants10.1016/j.actbio.2025.03.025 ↗
28. Biodegradable Zn-Li-Mn alloy to achieve optimal strength and ductility for bone implants10.1016/j.actbio.2025.04.056 ↗
29. Biodegradable Zn–Mn–Li alloy with a promising balance of mechanical property and corrosion resistance10.1016/j.jmrt.2024.03.202 ↗
30. Effect of exposure time on corrosion behavior of zinc-alloy in simulated body fluid solution: Electrochemical and surface investigation10.1016/j.jmrt.2020.12.050 ↗
31. Comparative Studies on Degradation Behavior of Pure Zinc in Various Simulated Body Fluids10.1007/s11837-019-03357-3 ↗
32. Hagelstein, S., Zankovic, S., Kovacs, A., Barkhoff, R., and Seidenstuecker, M. (2022). Mechanical Analysis and Corrosion Analysis of Zinc Alloys for Bioabsorbable Implants for Osteosynthesis. Materials, 15.10.3390/ma15020421 ↗
33. Improved biodegradability of zinc and its alloys by sandblasting treatment10.1016/j.surfcoat.2020.126678 ↗
34. The influence of alloying and fabrication techniques on the mechanical properties, biodegradability and biocompatibility of zinc: A comprehensive review10.1016/j.actbio.2019.01.035 ↗
35. Experimental study of corrosion rate supplied with an ab-initio elucidation of corrosion mechanism of biodegradable implants based on Ag-doped Zn alloys10.1016/j.apsusc.2024.159300 ↗
36. Mamrilla, W., Molcanova, Z., Ballokova, B., Dzupon, M., Dzunda, R., Csik, D., Michalik, S., Lisnichuk, M., and Saksl, K. (2023). The Influence of Manganese Addition on the Properties of Biodegradable Zinc-Manganese-Calcium Alloys. Materials, 16.10.3390/ma16134655 ↗
37. Yang, L., Li, X., Yang, L., Zhu, X., Wang, M., Song, Z., Liu, H.H., Sun, W., Dong, R., and Yue, J. (2023). Effect of Mg Contents on the Microstructure, Mechanical Properties and Cytocompatibility of Degradable Zn-0.5Mn-xMg Alloy. J. Funct. Biomater., 14.10.3390/jfb14040195 ↗
38. Enhancement of mechanical, corrosion, and biocompatibility properties of zinc alloys through magnesium and neodymium alloying for bone implant applications10.1039/D5RA07819A ↗
39. Liu, Q., Li, A., Liu, S., Fu, Q., Xu, Y., Dai, J., Li, P., and Xu, S. (2023). Cytotoxicity of Biodegradable Zinc and Its Alloys: A Systematic Review. J. Funct. Biomater., 14.10.3390/jfb14040206 ↗
40. Laser powder bed fusion: A state-of-the-art review of the technology, materials, properties & defects, and numerical modelling10.1016/j.jmrt.2022.07.121 ↗
41. Densification behavior of pure Zn metal parts produced by selective laser melting for manufacturing biodegradable implants10.1016/j.jmatprotec.2018.03.007 ↗
42. Laser Additive Manufacturing of Zinc Targeting for Biomedical Application10.18063/ijb.v8i1.501 ↗
43. Additively manufactured biodegradable Zn-Mn-based implants with an unprecedented balance of strength and ductility10.1016/j.actbio.2025.02.047 ↗
44. Biodegradability, biocompatibility, and mechanical behavior of additively manufactured zinc scaffolds10.1016/j.jmbbm.2024.106868 ↗
45. Recent progress and perspectives in laser additive manufacturing of biodegradable zinc alloy10.1016/j.jmrt.2024.11.012 ↗
46. Response of human periosteal cells to degradation products of zinc and its alloy10.1016/j.msec.2019.110208 ↗
47. The biological effects of copper alloying in Zn-based biodegradable arterial implants10.1016/j.bioadv.2024.214112 ↗
48. The role of Cu addition in the metallurgical features, mechanical properties, and cytocompatibility of cardiovascular stents biodegradable Zn-based alloy10.1016/j.intermet.2023.108106 ↗
49. Recent progress of novel biodegradable zinc alloys: From the perspective of strengthening and toughening10.1016/j.jmrt.2022.01.004 ↗
50. In vitro and in vivo studies of Zn-Mn biodegradable metals designed for orthopedic applications10.1016/j.actbio.2020.03.009 ↗
51. Improved mechanical and corrosion properties through high-pressure phase transition in a biodegradable Zn-1.5Mn alloy10.1016/j.corsci.2024.112399 ↗
52. Effect of Mn content on the corrosion behavior and biocompatibility of biodegradable Zn-Mn alloys10.1038/s41598-025-93296-8 ↗
53. A biodegradable Zn-1Cu-0.1Ti alloy with antibacterial properties for orthopedic applications10.1016/j.actbio.2020.02.017 ↗
54. Multi-objective optimization of LPBF manufacturing with Zn-4Al-1Cu alloy for technical applications10.1016/j.jmapro.2024.12.049 ↗
55. (2012). Biological Evaluation of Medical Devices—Part 12: Sample Preparation and Reference Materials (Standard No. ISO 10993-12:2012).
56. A new method for the model-independent assessment of thickness in three-dimensional images10.1046/j.1365-2818.1997.1340694.x ↗
57. Validation of corrosion rates measured by the Tafel extrapolation method10.1016/j.corsci.2005.05.046 ↗
58. (2026, March 10). Electrochemistry and Corrosion: Overview and Techniques. Princeton Applied Research Application Note CORR-4, Available online: https://de.scribd.com/document/341864672/Application-Note-CORR-4-pdf.
59. In vitro biodegradation behavior, mechanical properties, and cytotoxicity of biodegradable Zn-Mg alloy10.1002/jbm.b.33341 ↗
60. Wang, Z., Wang, W., Zhang, X., Cao, F., Zhang, T., Bhakta Pokharel, D., Chen, D., Li, J., Yang, J., and Xiao, C. (2022). Modulation of Osteogenesis and Angiogenesis Activities Based on Ionic Release from Zn-Mg Alloys. Materials, 15.10.3390/ma15207117 ↗
61. In vitro cytocompatibility and antibacterial studies on biodegradable Zn alloys supplemented by a critical assessment of direct contact cytotoxicity assay10.1002/jbm.b.35147 ↗
62. Microstructure refinement in biodegradable Zn-Cu-Ca alloy for enhanced mechanical properties, degradation homogeneity, and strength retention in simulated physiological condition10.1016/j.jmst.2022.03.006 ↗
63. Mechanical properties, in vitro degradation behavior, hemocompatibility and cytotoxicity evaluation of Zn–1.2Mg alloy for biodegradable implants10.1039/C6RA14300H ↗
64. Evaluation of as-extruded ternary Zn–Mg–Zr alloys for biomedical implantation material: In vitro and in vivo behavior10.1002/maco.201810648 ↗
65. Biological properties of Zn–0.04Mg–2Ag: A new degradable zinc alloy scaffold for repairing large-scale bone defects10.1016/j.jmrt.2021.05.096 ↗
66. Biocompatibility and biodegradation studies of a commercial zinc alloy for temporary mini-implant applications10.1038/s41598-017-15873-w ↗
67. Mechanical Strength, Biodegradation, and in Vitro and in Vivo Biocompatibility of Zn Biomaterials10.1021/acsami.8b20634 ↗
68. Effects of Li addition on the properties of biodegradable Zn–Fe–Li alloy: Microstructure, mechanical properties, corrosion behavior, and cytocompatibility10.1016/j.mtcomm.2024.108661 ↗
69. Potential biodegradable Zn-Cu binary alloys developed for cardiovascular implant applications10.1016/j.jmbbm.2017.05.013 ↗
70. Verifying the cytotoxicity of a biodegradable zinc alloy with nanodiamond sensors10.1016/j.bioadv.2024.213927 ↗
71. Mechanical properties and cytocompatibility of dense and porous Zn produced by laser powder bed fusion for biodegradable implant applications10.1016/j.actbio.2020.04.006 ↗
72. Introducing novel bioabsorbable Zn–Ag–Mg alloys intended for cardiovascular applications10.1016/j.mtcomm.2023.105544 ↗
73. Structure, mechanical characteristics and in vitro degradation, cytotoxicity, genotoxicity and mutagenicity of novel biodegradable Zn-Mg alloys10.1016/j.msec.2015.08.015 ↗
74. Corrosion behavior of biodegradable metals in two different simulated physiological solutions: Comparison of Mg, Zn and Fe10.1016/j.corsci.2021.109278 ↗
75. Test conditions can significantly affect the results of in vitro cytotoxicity testing of degradable metallic biomaterials10.1038/s41598-021-85019-6 ↗
76. Selection of extraction medium influences cytotoxicity of zinc and its alloys10.1016/j.actbio.2019.03.013 ↗
77. Influence of surface pre-treatment on the cytocompatibility of a novel biodegradable ZnMg alloy10.1016/j.msec.2016.05.114 ↗
78. Albumins inhibit the corrosion of absorbable Zn alloys at initial stages of degradation10.1680/jsuin.19.00063 ↗
79. Double-edged effects and mechanisms of Zn2+ microenvironments on osteogenic activity of BMSCs: Osteogenic differentiation or apoptosis10.1039/D0RA01465F ↗
80. Recent research and progress of biodegradable zinc alloys and composites for biomedical applications: Biomechanical and biocorrosion perspectives
81. Shishir, R., Nasiruddin, U., Manojkumar, P., Lokeshkumar, E., Ponnilavan, V., Parfenov, E.V., Valiev, R.Z., and Rameshbabu, N. (Mater. Today Proc., 2023). Zinc as a potential bio-degradable temporary implant material, Mater. Today Proc., in press.10.1016/j.matpr.2023.09.009 ↗
82. Degradation of zinc in saline solutions, plasma, and whole blood10.1002/jbm.b.33458 ↗
83. High-toughness Zn-Mn-Ca alloys as potential materials for intestinal implants10.1016/j.jmst.2024.12.053 ↗
84. Mechanical and bio-corrosion properties of quaternary Mg–Ca–Mn–Zn alloys compared with binary Mg–Ca alloys10.1016/j.matdes.2013.06.055 ↗
85. Ben Tzion-Mottye, L., Ron, T., Eliezer, D., and Aghion, E. (2022). The Effect of Mn on the Mechanical Properties and In Vitro Behavior of Biodegradable Zn-2%Fe Alloy. Metals, 12.10.3390/met12081291 ↗
86. Biodegradable Zn-Cu-Mn alloy with suitable mechanical performance and in vitro degradation behavior as a promising candidate for vascular stents10.1016/j.msec.2022.112652 ↗
87. The oxidation of zinc in air studied by XPS and AES10.1116/1.572331 ↗
88. Katarivas Levy, G., Goldman, J., and Aghion, E. (2017). The Prospects of Zinc as a Structural Material for Biodegradable Implants—A Review Paper. Metals, 7.10.3390/met7100402 ↗
89. Fluid-induced corrosion behavior of degradable zinc for stent application10.1016/j.jmst.2021.02.050 ↗
90. Mechanical properties, corrosion and degradation behaviors, and in vitro cytocompatibility of a biodegradable Zn-5La alloy for bone-implant applications10.1016/j.actbio.2023.07.061 ↗
91. Interplay between biomaterials and the immune system: Challenges and opportunities in regenerative medicine10.1016/j.actbio.2022.11.003 ↗
92. A cross-linked coating loaded with antimicrobial peptides for corrosion control, early antibacterial, and sequential osteogenic promotion on a magnesium alloy as orthopedic implants10.1016/j.actbio.2024.12.046 ↗
93. Surface modification on biodegradable zinc alloys10.1016/j.jmrt.2023.06.149 ↗
94. Biocomposites-Coated Biodegradable Materials with Optimized Properties for Orthopedic Implant Biodegradability and Performance: A Comparative Study10.1021/acsabm.5c00603 ↗
95. Surface modification on Zn-based biodegradable metals: Recent advances and future perspectives10.1007/s44251-025-00101-9 ↗