Quantification of Circulating Cell-Free DNA as a NETosis Marker in Trauma Patients with Type 2 Diabetes Mellitus

Type 2 diabetes mellitus (T2DM) significantly impairs fracture healing, with neutrophils playing a crucial role in this process. In T2DM, these immune cells are over-activated, leading to the excessive release of neutrophil extracellular traps (NETs), increasing inflammation and hindering recovery....

Full description

Saved in:
Bibliographic Details
Main Authors: Filiz Sahin, Regina Breinbauer, Caren Linnemann, Melike Tombaz, Andreas K. Nussler, Sabrina Ehnert
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Methods and Protocols
Subjects:
Online Access:https://www.mdpi.com/2409-9279/8/2/42
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849714980107059200
author Filiz Sahin
Regina Breinbauer
Caren Linnemann
Melike Tombaz
Andreas K. Nussler
Sabrina Ehnert
author_facet Filiz Sahin
Regina Breinbauer
Caren Linnemann
Melike Tombaz
Andreas K. Nussler
Sabrina Ehnert
author_sort Filiz Sahin
collection DOAJ
description Type 2 diabetes mellitus (T2DM) significantly impairs fracture healing, with neutrophils playing a crucial role in this process. In T2DM, these immune cells are over-activated, leading to the excessive release of neutrophil extracellular traps (NETs), increasing inflammation and hindering recovery. Thus, a need for markers to assess patients in the risk group arises. This study demonstrates that circulating cell-free DNA (cfDNA) can be efficiently quantified from serum samples by a single-step qPCR and be used as a marker for NETosis. Our results revealed that trauma patients with T2DM have the highest cfDNA levels, followed by trauma patients, and the healthy group has the lowest. The method shows strong correlations between cfDNA and neutrophil-specific markers such as MPO, citH3, AZU1, and α-defensin, highlighting its potential as a rapid indicator of NETosis. This approach could allow the timely interference for high-risk patients, ultimately improving healing outcomes and reducing complications such as chronic inflammation, non-union fractures, and diabetic foot ulcers.
format Article
id doaj-art-3cc17bf9fdab45959b636bf718279b4e
institution DOAJ
issn 2409-9279
language English
publishDate 2025-04-01
publisher MDPI AG
record_format Article
series Methods and Protocols
spelling doaj-art-3cc17bf9fdab45959b636bf718279b4e2025-08-20T03:13:32ZengMDPI AGMethods and Protocols2409-92792025-04-01824210.3390/mps8020042Quantification of Circulating Cell-Free DNA as a NETosis Marker in Trauma Patients with Type 2 Diabetes MellitusFiliz Sahin0Regina Breinbauer1Caren Linnemann2Melike Tombaz3Andreas K. Nussler4Sabrina Ehnert5Siegfried Weller Institute for Trauma Research, BG Unfallklinik Tübingen, Eberhard Karls Universität Tuebingen, 72076 Tuebingen, GermanySiegfried Weller Institute for Trauma Research, BG Unfallklinik Tübingen, Eberhard Karls Universität Tuebingen, 72076 Tuebingen, GermanySiegfried Weller Institute for Trauma Research, BG Unfallklinik Tübingen, Eberhard Karls Universität Tuebingen, 72076 Tuebingen, GermanySiegfried Weller Institute for Trauma Research, BG Unfallklinik Tübingen, Eberhard Karls Universität Tuebingen, 72076 Tuebingen, GermanySiegfried Weller Institute for Trauma Research, BG Unfallklinik Tübingen, Eberhard Karls Universität Tuebingen, 72076 Tuebingen, GermanySiegfried Weller Institute for Trauma Research, BG Unfallklinik Tübingen, Eberhard Karls Universität Tuebingen, 72076 Tuebingen, GermanyType 2 diabetes mellitus (T2DM) significantly impairs fracture healing, with neutrophils playing a crucial role in this process. In T2DM, these immune cells are over-activated, leading to the excessive release of neutrophil extracellular traps (NETs), increasing inflammation and hindering recovery. Thus, a need for markers to assess patients in the risk group arises. This study demonstrates that circulating cell-free DNA (cfDNA) can be efficiently quantified from serum samples by a single-step qPCR and be used as a marker for NETosis. Our results revealed that trauma patients with T2DM have the highest cfDNA levels, followed by trauma patients, and the healthy group has the lowest. The method shows strong correlations between cfDNA and neutrophil-specific markers such as MPO, citH3, AZU1, and α-defensin, highlighting its potential as a rapid indicator of NETosis. This approach could allow the timely interference for high-risk patients, ultimately improving healing outcomes and reducing complications such as chronic inflammation, non-union fractures, and diabetic foot ulcers.https://www.mdpi.com/2409-9279/8/2/42neutrophil extracellular trapstraumadiabetes mellitusNET marker
spellingShingle Filiz Sahin
Regina Breinbauer
Caren Linnemann
Melike Tombaz
Andreas K. Nussler
Sabrina Ehnert
Quantification of Circulating Cell-Free DNA as a NETosis Marker in Trauma Patients with Type 2 Diabetes Mellitus
Methods and Protocols
neutrophil extracellular traps
trauma
diabetes mellitus
NET marker
title Quantification of Circulating Cell-Free DNA as a NETosis Marker in Trauma Patients with Type 2 Diabetes Mellitus
title_full Quantification of Circulating Cell-Free DNA as a NETosis Marker in Trauma Patients with Type 2 Diabetes Mellitus
title_fullStr Quantification of Circulating Cell-Free DNA as a NETosis Marker in Trauma Patients with Type 2 Diabetes Mellitus
title_full_unstemmed Quantification of Circulating Cell-Free DNA as a NETosis Marker in Trauma Patients with Type 2 Diabetes Mellitus
title_short Quantification of Circulating Cell-Free DNA as a NETosis Marker in Trauma Patients with Type 2 Diabetes Mellitus
title_sort quantification of circulating cell free dna as a netosis marker in trauma patients with type 2 diabetes mellitus
topic neutrophil extracellular traps
trauma
diabetes mellitus
NET marker
url https://www.mdpi.com/2409-9279/8/2/42
work_keys_str_mv AT filizsahin quantificationofcirculatingcellfreednaasanetosismarkerintraumapatientswithtype2diabetesmellitus
AT reginabreinbauer quantificationofcirculatingcellfreednaasanetosismarkerintraumapatientswithtype2diabetesmellitus
AT carenlinnemann quantificationofcirculatingcellfreednaasanetosismarkerintraumapatientswithtype2diabetesmellitus
AT meliketombaz quantificationofcirculatingcellfreednaasanetosismarkerintraumapatientswithtype2diabetesmellitus
AT andreasknussler quantificationofcirculatingcellfreednaasanetosismarkerintraumapatientswithtype2diabetesmellitus
AT sabrinaehnert quantificationofcirculatingcellfreednaasanetosismarkerintraumapatientswithtype2diabetesmellitus