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Article

Extracellular Vesicle Isolation and Characterization from Periprosthetic Joint Synovial Fluid in Revision Total Joint Arthroplasty

1
Clinic for Orthopedics and Trauma Surgery, University Hospital Bonn, 53127 Bonn, Germany
2
Electron Microscopy and Analytics, Center of Advanced European Studies and Research, 53175 Bonn, Germany
3
Medical Faculty, University Hospital Essen, 45147 Essen, Germany
4
Department of Orthopaedics and Trauma Surgery, University Medical Center Mannheim of University Heidelberg, 68167 Mannheim, Germany
*
Author to whom correspondence should be addressed.
J. Clin. Med. 2020, 9(2), 516; https://doi.org/10.3390/jcm9020516
Received: 15 January 2020 / Revised: 7 February 2020 / Accepted: 11 February 2020 / Published: 14 February 2020
(This article belongs to the Special Issue Advances in Prosthetic Joint Infection Care)
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. View Full-Text
Keywords: EVs; extracellular vesicle isolation; periprosthetic joint synovial fluid; periprosthetic joint infection; aseptic implant failure EVs; extracellular vesicle isolation; periprosthetic joint synovial fluid; periprosthetic joint infection; aseptic implant failure
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MDPI and ACS Style

Rüwald, J.M.; Randau, T.M.; Hilgers, C.; Masson, W.; Irsen, S.; Eymael, R.L.; Kohlhof, H.; Gravius, S.; Burger, C.; Wirtz, D.C.; Schildberg, F.A. Extracellular Vesicle Isolation and Characterization from Periprosthetic Joint Synovial Fluid in Revision Total Joint Arthroplasty. J. Clin. Med. 2020, 9, 516. https://doi.org/10.3390/jcm9020516

AMA Style

Rüwald JM, Randau TM, Hilgers C, Masson W, Irsen S, Eymael RL, Kohlhof H, Gravius S, Burger C, Wirtz DC, Schildberg FA. Extracellular Vesicle Isolation and Characterization from Periprosthetic Joint Synovial Fluid in Revision Total Joint Arthroplasty. Journal of Clinical Medicine. 2020; 9(2):516. https://doi.org/10.3390/jcm9020516

Chicago/Turabian Style

Rüwald, Julian M.; Randau, Thomas M.; Hilgers, Cäcilia; Masson, Werner; Irsen, Stephan; Eymael, Robin L.; Kohlhof, Hendrik; Gravius, Sascha; Burger, Christof; Wirtz, Dieter C.; Schildberg, Frank A. 2020. "Extracellular Vesicle Isolation and Characterization from Periprosthetic Joint Synovial Fluid in Revision Total Joint Arthroplasty" J. Clin. Med. 9, no. 2: 516. https://doi.org/10.3390/jcm9020516

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