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Ionic silver coating of orthopedic implants may impair osteogenic differentiation and mineralization
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Surgical Sciences, Orthopaedics and Handsurgery.ORCID iD: 0000-0002-1502-1221
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Surgical Sciences, Orthopaedics and Handsurgery.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Surgical Sciences, Orthopaedics and Handsurgery.ORCID iD: 0000-0002-1591-3828
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Surgical Sciences, Orthopaedics and Handsurgery.ORCID iD: 0000-0002-3233-2638
2025 (English)In: Experimental and Therapeutic Medicine, ISSN 1792-0981, E-ISSN 1792-1015, Vol. 29, no 3, article id 51Article in journal (Refereed) Published
Abstract [en]

Silver (Ag) possesses potent antimicrobial properties and is used as a coating for medical devices. The impact of silver ions released from orthopedic implants on the differentiation and osteoid formation of different osteogenic cells has yet to be systematically studied. In the present study, human mesenchymal stem cells (hMSCs) and primary human osteoblasts (hOBs) were exposed to different static Ag+ concentrations (0, 0.5, 1.0 or 1.5 ppm) or dynamic Ag+ concentrations (range 0 to 0.7 ppm) that simulated the temporal release pattern from a Ag‑nitrate coating of trabecular titanium (TLSN). Cell morphology was investigated by phase contrast and fluorescence microscopy. The activities of alkaline phosphatase (ALP) and lactate dehydrogenase, osteogenic gene expression (COL1A1, COL1A2 and ALPL), and osteoid deposition were examined for up to 4 weeks. DAPI and carboxyfluorescein diacetate staining revealed changes in the morphology of hOBs treated with ≥0.5 ppm Ag+, while osteocalcin‑positive cells were observed primarily in the untreated group. Elevated Ag+ concentrations did not impact the production of ALP by either hMSCs or hOBs. Treatment with 1.5 ppm Ag+ or TLSN Ag+ led to a modest reduction in COL1A2 and ALPL levels in hMSCs at 2 weeks but not at 4 weeks nor in hOBs. In hMSC cultures, mineralization decreased at ≥1 ppm Ag+, whereas the same concentration range significantly reduced mineralization in hOB cultures. In conclusion, Ag+ concentrations ranging from 1.0 to 1.5 ppm may interfere with osteogenic differentiation, possibly by altering gene expression, thereby affecting mineralization. Only Ag+ concentrations up to 0.5 ppm allowed undisturbed osteogenic differentiation and mineralization. These findings pertain to creating Ag coatings of titanium intended for cementless fixation into host bone.

Place, publisher, year, edition, pages
Spandidos Publications , 2025. Vol. 29, no 3, article id 51
Keywords [en]
silver-coating, osseointegration, human osteoblasts, infection
National Category
Biomaterials Science Orthopaedics
Identifiers
URN: urn:nbn:se:uu:diva-550404DOI: 10.3892/etm.2025.12801ISI: 001408869200001PubMedID: 39885915OAI: oai:DiVA.org:uu-550404DiVA, id: diva2:1938620
Funder
Swedish Research Council, 2021-00980Promobilia foundation, A23003Available from: 2025-02-19 Created: 2025-02-19 Last updated: 2025-02-23Bibliographically approved
In thesis
1. Filling bone defects: Antibacterial titanium implants and 3D-printed bone
Open this publication in new window or tab >>Filling bone defects: Antibacterial titanium implants and 3D-printed bone
2025 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This work, comprising four studies, focuses on experimental implants and materials for orthopaedic applications designed to address the often concomitant issues of infections and bone loss. The first two studies examine silver (Ag) as an antibacterial agent and its effects on human cells when used as a coating on titanium (Ti) implants. The last two studies explore methods for regenerating bone defects.

In the first study, we compared two types of Ag-coated Ti implants provided by an implant manufacturer. One implant is used clinically, while the other was experimental and contained smaller amounts of Ag. Physicochemical analysis showed that the entire surface of the clinical implant was coated with Ag, while the Ag on the experimental implant formed aggregates on the surface. The clinically used implant released significantly more Ag, while, the experimental implant's release ceased after a few days. S. aureus and osteoblasts was cultured separately then on the implants, and we showed that smaller amounts of Ag on the implants maintained satisfactory antibacterial effects while minimising adverse effects on the osteoblasts.

The second study investigated the effects of different ionic Ag concentrations on osteoblasts and mesenchymal stem cells. Using various methods, including PCR, enzymatic analyses for cell differentiation, microscopy, and staining for mineralisation, we found that even small amounts of Ag could inhibit mineralisation by human osteoblasts. We observed no significant impact on the osteogenic differentiation of osteoblasts or mesenchymal stem cells regarding gene expression and ALP production. However, microscopic analysis revealed abnormal cell patterns, such as reduced confluence in Ag-treated groups compared to controls. These findings, suggest that Ag-coated implants should be used cautiously in clinical settings, especially in parts of the prosthesis intended for direct bone integration.

Study III employed the freeform reversible embedding of suspended hydrogels (FRESH) technique, to 3D-print collagen structures with trabecular geometry. This technique and its structures were compared to stereolithographic printing, an alternative 3D printing method, which produced analogous structures using a different material, polyethylene glycol diacrylate (PEGDA). Rheological and mechanical analyses were performed on the 3D-printed collagen structures to characterise their elasticity and stiffness; electron microscopy was used to map the surface geometry of the collagen and PEGDA structures. Cultures of human osteoblasts and osteosarcoma cell lines on the two types of structures showed that the 3D-printed collagen structures were better suited as carriers for osteogenic cells compared to the 3D-printed PEGDA structures.

In the fourth study, we successfully 3D-printed artificial bone of collagen and hydroxyapatite. The collagen in this material was modified by cross-linking. Mechanical and rheological analyses of the material in gel and solid forms were performed, and the distribution pattern of hydroxyapatite within the material was examined. Finally, we cultured human osteoblasts on the 3D-printed collagen-hydroxyapatite structures up to four weeks. We found that osteoblasts grew well and differentiated satisfactorily on structures printed with our new material.

Overall, this work shows how surface coatings and 3D printing might solve the two biggest issues in orthopaedics—infections and missing bone. We trust this dissertation will illuminate the underlying scientific principles of these techniques for orthopaedic surgeons.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2025. p. 81
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 2126
Keywords
antibacterial coatings, bone tissue engineering, 3D printing, bone defects, orthopaedics
National Category
Orthopaedics
Identifiers
urn:nbn:se:uu:diva-551294 (URN)978-91-513-2399-2 (ISBN)
Public defence
2025-04-11, Rosénsalen ing 95/96, Akademiska Sjukhuset, Sjukhusvägen, Uppsala, 09:00 (English)
Opponent
Supervisors
Available from: 2025-03-21 Created: 2025-02-23 Last updated: 2025-03-21

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