title: Wear particles derived from metal hip implants induce the generation of multinucleated giant cells in a 3-dimensional peripheral tissue-equivalent model.
abstract: Multinucleate giant cells (MGCs) are formed by the fusion of 5 to 15 monocytes or macrophages. MGCs can be generated by hip implants at the site where the metal surface of the device is in close contact with tissue. MGCs play a critical role in the inflammatory processes associated with adverse events such as aseptic loosening of the prosthetic joints and bone degeneration process called osteolysis. Upon interaction with metal wear particles, endothelial cells upregulate pro-inflammatory cytokines and other factors that enhance a localized immune response. However, the role of endothelial cells in the generation of MGCs has not been completely investigated. We developed a three-dimensional peripheral tissue-equivalent model (PTE) consisting of collagen gel, supporting a monolayer of endothelial cells and human peripheral blood mononuclear cells (PBMCs) on top, which mimics peripheral tissue under normal physiological conditions. The cultures were incubated for 14 days with Cobalt chromium alloy (CoCr ASTM F75, 1-5 micron) wear particles. PBMC were allowed to transit the endothelium and harvested cells were analyzed for MGC generation via flow cytometry. An increase in forward scatter (cell size) and in the propidium iodide (PI) uptake (DNA intercalating dye) was used to identify MGCs. Our results show that endothelial cells induce the generation of MGCs to a level 4 fold higher in 3-dimentional PTE system as compared to traditional 2-dimensional culture plates. Further characterization of MGCs showed upregulated expression of tartrate resistant alkaline phosphatase (TRAP) and dendritic cell specific transmembrane protein, (DC-STAMP), which are markers of bone degrading cells called osteoclasts. In sum, we have established a robust and relevant model to examine MGC and osteoclast formation in a tissue like environment using flow cytometry and RT-PCR. With endothelial cells help, we observed a consistent generation of metal wear particle- induced MGCs, which heralds metal on metal hip failures.
authors.affiliation: Department of Biology, Chemistry and Materials Science, Office of Science and Engineering Laboratories, CDRH, FDA, Silver Spring, Maryland, United States of America; Department of Medicine, Uniformed Services University of the Health Sciences, Bethesda, Maryland, United States of America.
authors.forename: Debargh K
authors.initials: DK
authors.lastname: Dutta
authors.affiliation: Department of Biology, Chemistry and Materials Science, Office of Science and Engineering Laboratories, CDRH, FDA, Silver Spring, Maryland, United States of America.
authors.forename: Pushya A
authors.initials: PA
authors.lastname: Potnis
authors.affiliation: Department of Biology, Chemistry and Materials Science, Office of Science and Engineering Laboratories, CDRH, FDA, Silver Spring, Maryland, United States of America; University of Maryland, College Park, Maryland, United States of America.
authors.forename: Kelly
authors.initials: K
authors.lastname: Rhodes
authors.affiliation: Department of Biology, Chemistry and Materials Science, Office of Science and Engineering Laboratories, CDRH, FDA, Silver Spring, Maryland, United States of America.
authors.forename: Steven C
authors.initials: SC
authors.lastname: Wood
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D002858:Chromium Alloys / Q000009:adverse effects*; D003343:Corrosion; D004730:Endothelium, Vascular / Q000187:drug effects / Q000473:pathology; D005434:Flow Cytometry; D005547:Foreign Bodies / Q000503:physiopathology*; D015726:Giant Cells / Q000187:drug effects / Q000473:pathology*; D006622:Hip Prosthesis / Q000009:adverse effects*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000473:pathology; D008264:Macrophages / Q000187:drug effects / Q000473:pathology; D008670:Metals / Q000009:adverse effects* / Q000737:chemistry; D009000:Monocytes / Q000187:drug effects / Q000473:pathology; D010010:Osteoclasts / Q000187:drug effects / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D002858:Chromium Alloys; D008670:Metals; D012333:RNA, Messenger