Metaproteomic analysis of human sonicate fluid highlights neutrophil extracellular trap formation associated with periprosthetic joint infection
5 affiliations
- 1Graduate School of Biomedical Sciences, Mayo Clinic, Rochester, Minnesota, USA
- 2Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota, USA
- 3Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota, USA
- 4Department of Quantitative Health Sciences, Mayo Clinic, Rochester, Minnesota, USA
- 5Physiology and Biomedical Engineering, Mayo Clinic, Rochester, Minnesota, USA
Abstract
Periprosthetic joint infection (PJI) is a devastating complication of total joint arthroplasty that can be challenging to diagnose and treat. Neutrophils are a prominent feature in PJI, with neutrophil extracellular traps (NETs) known to be present; little is known, however, about the physiology of PJI-associated NETs. Here, the proteomes of PJI and non-infectious arthroplasty failure (NIAF), analyzed using liquid chromatography tandem mass-spectrometry (LC-MS/MS) and proximity extension assay (PEA)-based profiling of fluids derived from sonication of arthroplasty components removed from patients who had undergone revision arthroplasty, were compared, with a focus on NET formation. In addition, in vitro effects of synovial fluid components on Staphylococcus aureus-induced NET formation were characterized. LC-MS/MS (PJI n = 85, NIAF n = 26) and PEA (PJI n = 95, NIAF n = 39) showed NET formation and neutrophil degranulation to be associated with PJI, with the complement cascade and platelet activation pathways identified as potential contributors. In vitro studies showed delayed NET formation in artificial synovial fluid compared to standard cell culture media conventionally used to evaluate NETs. Together, these results demonstrate the presence of NETs in PJI, identify potential pathways involved in PJI-associated NETosis, and point to an influence of synovial components on NET formation during PJI.IMPORTANCEPeriprosthetic joint infection (PJI) is a major complication of joint replacement, placing a significant burden on patients and the healthcare system. Our research reveals that neutrophils and their antimicrobial “traps” are dominant features of the immune response at the infection joint site. Importantly, we found that the unique environment of the joint delays the formation of these traps, potentially contributing to the pathogenesis of PJI. By establishing a new experimental platform tailored for the infected joint, this work provides a foundational platform for probing these mechanisms and supports the development of improved diagnostic and therapeutic strategies.
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BibTeX
@article{garcia2026metaproteomic,
title = {{Metaproteomic analysis of human sonicate fluid highlights neutrophil extracellular trap formation associated with periprosthetic joint infection}},
author = {Elijah Garcia and Kathleen O'Connor and Christina Koscianski and Judith Alvarez Otero and Keara O’Donnell and Kiran K. Mangalaparthi and Nicholas B. Larson and Akhilesh Pandey and David Holmes and Robin Patel},
journal = {Microbiology Spectrum},
year = {2026},
month = nov,
doi = {10.1128/spectrum.00665-26},
url = {https://doi.org/10.1128/spectrum.00665-26},
note = {CC BY},
}
RIS
TY - JOUR
TI - Metaproteomic analysis of human sonicate fluid highlights neutrophil extracellular trap formation associated with periprosthetic joint infection
AU - Elijah Garcia
AU - Kathleen O'Connor
AU - Christina Koscianski
AU - Judith Alvarez Otero
AU - Keara O’Donnell
AU - Kiran K. Mangalaparthi
AU - Nicholas B. Larson
AU - Akhilesh Pandey
AU - David Holmes
AU - Robin Patel
PY - 2026
DA - 2026/11/03
JO - Microbiology Spectrum
DO - 10.1128/spectrum.00665-26
UR - https://doi.org/10.1128/spectrum.00665-26
AB - Periprosthetic joint infection (PJI) is a devastating complication of total joint arthroplasty that can be challenging to diagnose and treat. Neutrophils are a prominent feature in PJI, with neutrophil extracellular traps (NETs) known to be present; little is known, however, about the physiology of PJI-associated NETs. Here, the proteomes of PJI and non-infectious arthroplasty failure (NIAF), analyzed using liquid chromatography tandem mass-spectrometry (LC-MS/MS) and proximity extension assay (PEA)-based profiling of fluids derived from sonication of arthroplasty components removed from patients who had undergone revision arthroplasty, were compared, with a focus on NET formation. In addition, in vitro effects of synovial fluid components on Staphylococcus aureus-induced NET formation were characterized. LC-MS/MS (PJI n = 85, NIAF n = 26) and PEA (PJI n = 95, NIAF n = 39) showed NET formation and neutrophil degranulation to be associated with PJI, with the complement cascade and platelet activation pathways identified as potential contributors. In vitro studies showed delayed NET formation in artificial synovial fluid compared to standard cell culture media conventionally used to evaluate NETs. Together, these results demonstrate the presence of NETs in PJI, identify potential pathways involved in PJI-associated NETosis, and point to an influence of synovial components on NET formation during PJI.IMPORTANCEPeriprosthetic joint infection (PJI) is a major complication of joint replacement, placing a significant burden on patients and the healthcare system. Our research reveals that neutrophils and their antimicrobial “traps” are dominant features of the immune response at the infection joint site. Importantly, we found that the unique environment of the joint delays the formation of these traps, potentially contributing to the pathogenesis of PJI. By establishing a new experimental platform tailored for the infected joint, this work provides a foundational platform for probing these mechanisms and supports the development of improved diagnostic and therapeutic strategies.
SN - 2165-0497
ER -