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Evaluation of a Fluid-based Screening Tool for Diagnosis of Heterotopic Ossification Using Cytokine Profiles
Hsiao Hsin Sung, DDS, Ron Allen, MS, Kavitha Ranganathan, MD, Shailesh Agarwal, MD, Shawn Loder, BS, James Drake, BS, David Cholok, BS, John Li, MD, Shuli Li, PhD, Yuji Mishina, PhD, Benjamin Levi, MD.
University of Michigan, Ann Arbor, MI, USA.

PURPOSE: Heterotopic ossification (HO) occurs in the setting of persistent systemic inflammation. Currently, we lack the ability to predict patients at risk for HO. The goal of this study is to develop a bedside-point-of-care assay to predict those at risk to develop HO, allowing more directed interventions.
METHODS: Serum from C57Bl/6 non-injured and injured mice after Achilles’ tenotomy were collected prior to the injury, and 48 hours and 3 weeks after injury. Serum from burn patients prior and after HO and a healthy control were also collected. Samples were analyzed for 19 cytokines by Bio-Plex-cytokines-Biorad and reassessed using ELISA-assays. The mRNA levels of these cytokines in the injured site were quantified with real-time-PCR.
RESULTS: Mouse MCP-1, IL-1β and IL-6 levels increased 3.7; 1.5 and 1.1 fold change (FC) after the injury. The increase in mice TNF-α was 1.23 FC and it is correlated with the timing of HO formation at 3 weeks post-injury. The level of mice TGF-β increased 1.3 and 1.96 FC at 48 hours and 3 weeks and was associated with an increase of ALP and Runx2 mRNA expression (p<0.005) at the tenotomy site. Human MCP-1 and MMP9 levels are higher in the pre-HO stage compared to the control (1 and 1.6 FC) or post-HO stage (123 and 7.9 FC, respectively).
CONCLUSIONS: In this study, we characterized the diagnostic potential of specific cytokines that can serve as biomarkers for an early stage diagnosis of HO. These findings support a possible correlation between serum cytokine levels and HO formation.


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