Elena Della Bella, Ursula Menzel, Andreas Naros, Eva Johanna Kubosch, Mauro Alini, Martin J. Stoddart.
Abstract
Fracture non-unions affect many patients worldwide, however, known risk factors alone do not predict individual risk. The identification of novel biomarkers is crucial for early diagnosis and timely patient treatment. This study focused on the identification of microRNA (miRNA) related to the process of fracture healing. Serum of fracture patients and healthy volunteers was screened by RNA sequencing to identify differentially expressed miRNA at various times after injury.
Introduction
Fracture non-unions affect many patients leading to functional and psychosocial disability. Several risk factors have been described and include biological, surgical, and mechanical factors. However, none of them are able to determine if healing abnormalities will arise. As a consequence, there are no prognostic markers to monitor fracture healing, leaving no option but to wait until a non-union has occurred. Bone turnover markers have some utility but there are still uncertainties in their clinical use for monitoring fracture healing.
Materials and Methods:
Whole blood was collected with full ethical approval and signed informed patient consent (EK-Freiburg 105/17 and ZH BASEC-Nr.2017-01390) from 10 healthy volunteers and 12 fracture patients (S1 Table). From fracture patients, blood was collected at day 3 (n = 10, "early"), between day 5 and 12 (n = 5, "late"), and between day 19 and 56 (n = 6, "very late") after fracture. After clotting, samples were centrifuged for 10 min at 2500g and 4°C. The serum was then filtered and frozen at –80°C.
Discussion
The identification of biomarkers to stratify patients’ non-union risk is crucial to allow early intervention. Blood-based biomarkers would be ideal since they can be obtained following a minimally invasive procedure and can be easily combined with the analysis of other biochemical parameters. To be robust, the marker must be present in a broad range of patients, for this reason we utilized an unmatched patient population. Despite this, we successfully identified potential markers and more clearly defined the time period when sample present the greatest number of miRNA changes.
Citation: Della Bella E, Menzel U, Naros A, Kubosch EJ, Alini M, Stoddart MJ (2024) Identification of circulating miRNAs as fracture-related biomarkers. PLoS ONE 19(5): e0303035. https://doi.org/10.1371/journal.pone.0303035
Editor: Carlos Alberto Antunes Viegas, Universidade de Trás-os-Montes e Alto Douro: Universidade de Tras-os-Montes e Alto Douro, PORTUGAL
Received: May 12, 2023; Accepted: April 16, 2024; Published: May 31, 2024.
Copyright: © 2024 Bella et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Data Availability: Sequencing data deposited in NCBI’s Gene Expression Omnibus with accession number GSE217879. The minimal underlying data set is available as supplementary file.
Funding: MJS and EDB obtained funding from AO Foundation and AO Trauma. https://www.aofoundation.org/what-we-do/research-innovation/about https://www.aofoundation.org/trauma The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
