RAW 264.7 co-cultured with ultra-high molecular weight polyethylene particles spontaneously differentiate into osteoclasts: an in vitro model of periprosthetic osteolysis.
Wear-particle osteolysis affects prosthesis survival leading to implant loosening up to 70% of revisions. Therapeutic strategies are increasing, however alternative testing methods to experimentally evaluate such treatments are lacking. The aim of this study was to reproduce an in vitro osteolysis model recapitulating the events that, starting from the exposure of macrophages to polyethylene, lead to the establishment of osteoclastogenesis and inflammation. Responses to polyethylene, at 3 and 7 days, in a macrophage cell line, RAW 264.7, were determined by DNA quantification, immunofluorescence, pit assay, gene expression, cytokine production and NF-kB activation. Results showed that 3 days exposure to particles could induce a significant production of Tumor Necrosis Factor alpha (p < 0.0005) and Prostaglandin E2(p < 0.005) compared to controls. Particles also induced macrophages to spontaneously differentiate into mature and active osteoclasts, in terms of identification of multinucleated cells by Phalloidin staining and by the analysis of osteoclast-specific gene markers. In particular, at 3 days polyethylene induced a significant up-regulation of Nuclear Factor of Activated T-cells, cytoplasmic 1, Receptor Activator of Nuclear factor Kappa-B and Receptor Activator of Nuclear Factor Kappa-B Ligand genes (p < 0.0005) compared to controls. At protein level, the particles induced a significant increase of Receptor Activator of Nuclear Factor Kappa-B Ligand at day 7 over controls (p < 0.0005). Osteoclasts were capable to resorb bone even in absence of differentiating factors. The possible mechanism, beside spontaneous osteoclastogenesis mediated by wear debris, was identified in an autocrine up-regulation of Receptor activator of nuclear factor kappa-B ligand gene expression and protein synthesis. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 510-520, 2017.
Laboratory of Biocompatibility, Technological Innovations and Advanced Therapies, BITTA, Rizzoli Orthopaedic Institute- Research, Innovation and Technology Department (RIT), via di Barbiano 1/10, 40136, Bologna, Italy.
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Sartori
Laboratory of Cellular and Molecular Pharmacology Department of Medical Sciences, University of Ferrara, via Fossato di Mortara 17-19, Ferrara, 44121, Italy.
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Vincenzi
Laboratory of Cellular and Molecular Pharmacology Department of Medical Sciences, University of Ferrara, via Fossato di Mortara 17-19, Ferrara, 44121, Italy.
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Ravani
Laboratory of Preclinical and Surgical Studies, Rizzoli Orthopaedic Institute, via di Barbiano 1/10, Bologna, 40136, Italy.
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Cepollaro
Laboratory of Preclinical and Surgical Studies, Rizzoli Orthopaedic Institute, via di Barbiano 1/10, Bologna, 40136, Italy.
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Martini
Laboratory of Cellular and Molecular Pharmacology Department of Medical Sciences, University of Ferrara, via Fossato di Mortara 17-19, Ferrara, 44121, Italy.
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Varani
Laboratory of Preclinical and Surgical Studies, Rizzoli Orthopaedic Institute, via di Barbiano 1/10, Bologna, 40136, Italy.
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Fini
Laboratory of Preclinical and Surgical Studies, Rizzoli Orthopaedic Institute, via di Barbiano 1/10, Bologna, 40136, Italy.
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Tschon
D000818:Animals; D000943:Antigens, Differentiation / Q000096:biosynthesis; D000069343:Bone-Implant Interface; D018920:Coculture Techniques; D051379:Mice; D010010:Osteoclasts / Q000378:metabolism* / Q000473:pathology; D010014:Osteolysis / Q000378:metabolism* / Q000473:pathology; D011095:Polyethylenes / Q000737:chemistry / Q000494:pharmacology; D000067996:RAW 264.7 Cells; D013997:Time Factors
D000943:Antigens, Differentiation; D011095:Polyethylenes; C111601:ultra-high molecular weight polyethylene
alternative in vitro model; macrophages; periprosthetic osteolysis; spontaneous osteoclastogenesis; ultra-high molecular weight polyethylene