Abstract
Extracellular vesicles (EVs) comprise an as yet insufficiently investigated intercellular communication pathway in the field of revision total joint arthroplasty (RTJA). This study examined whether periprosthetic joint synovial fluid contains EVs, developed a protocol for their isolation and characterized them with respect to quantity, size, surface markers as well as documented their differences between aseptic implant failure (AIF) and periprosthetic joint infection (PJI). EV isolation was accomplished using ultracentrifugation, electron microscopy (EM) and nanoparticle tracking analysis evaluated EV presence as well as particle size and quantity. EV surface markers were studied by a bead-based multiplex analysis. Using our protocol, EM confirmed the presence of EVs in periprosthetic joint synovial fluid. Higher EV particle concentrations and decreased particle sizes were apparent for PJI. Multiplex analysis confirmed EV-typical surface epitopes and revealed upregulated CD44 and HLA-DR/DP/DQ for AIF, as well as increased CD40 and CD105. Our protocol achieved isolation of EVs from periprosthetic joint synovial fluid, confirmed by EM and multiplex analysis. Characterization was documented with respect to size, concentration and epitope surface signature. Our results indicate various differences between PJI and AIF EVs. This pilot study enables new research approaches and rising diagnostic opportunities in the field of RTJA.
Highlights
The periprosthetic joint infection (PJI) is a frequent and devastating complication following knee or hip arthroplasty with dramatic impact on patients and healthcare systems [1,2,3]
Since the first description of exosomes in the 1980s as a vehicle to dispose cellular waste, scientific convergence has evolved to attribute enormous diagnostic potential to nanoscale vesicles as multimodal reflectors of a cell’sphysiological state [9,10]. These properties have dubbed extracellular vesicles (EVs) “liquid biopsies”, which are released into the extracellular space by multicellular organisms with enclosed proteins, lipids and nucleic acids as their cargo to act on distant receiver cells in form of exosomes (30–100 nm) and microvesicles (50–1000 nm) or, in case of cell death, apoptotic bodies (1000–5000 nm) [11,12]
Immune regulatory functions are an established property of EVs and it has been shown that bacterial infections can elicit the immunologic release of nanovesicles with distinct molecular characteristics [17,18,19]
Summary
The periprosthetic joint infection (PJI) is a frequent and devastating complication following knee or hip arthroplasty with dramatic impact on patients and healthcare systems [1,2,3]. Since the first description of exosomes in the 1980s as a vehicle to dispose cellular waste, scientific convergence has evolved to attribute enormous diagnostic potential to nanoscale vesicles as multimodal reflectors of a cell’s (patho-)physiological state [9,10]. These properties have dubbed EVs “liquid biopsies”, which are released into the extracellular space by multicellular organisms with enclosed proteins, lipids and nucleic acids as their cargo to act on distant receiver cells in form of exosomes (30–100 nm) and microvesicles (50–1000 nm) or, in case of cell death, apoptotic bodies (1000–5000 nm) [11,12]. This study aims to isolate and characterize EVs from periprosthetic joint aspirates with respect to their quantity, size, surface epitopes as well as possible variations between PJI and AIF
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