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ORIGINAL RESEARCH article

Front. Bioeng. Biotechnol.

Sec. Biosafety and Biosecurity

Volume 13 - 2025 | doi: 10.3389/fbioe.2025.1617285

Effective Detection of Staphylococcal Infections in Human Bone Tissue using Combined Raman Microscopy and Micro-Computed Tomography

Provisionally accepted
Richard  Andreas LindtnerRichard Andreas Lindtner1Lukas  KampikLukas Kampik1Larissa  NoackLarissa Noack1David  PutzerDavid Putzer1Rohit  AroraRohit Arora1Débora  Coraça-HuberDébora Coraça-Huber1Michael  SchirmerMichael Schirmer1Gerald  DegenhartGerald Degenhart1Michaela  LacknerMichaela Lackner1Jürgen  PoppJürgen Popp2Jovan  BadzokaJovan Badzoka3Christoph  KappacherChristoph Kappacher3Christian  Wolfgang HuckChristian Wolfgang Huck3Johannes  Dominikus PalluaJohannes Dominikus Pallua1*
  • 1Innsbruck Medical University, Innsbruck, Tyrol, Austria
  • 2Leibniz Institute of Photonic Technology (IPHT), Jena, Thuringia, Germany
  • 3University of Innsbruck, Innsbruck, Tyrol, Austria

The final, formatted version of the article will be published soon.

Bone infections caused by Staphylococcus aureus and Staphylococcus epidermidis are serious complications in orthopedic surgery. These infections commonly occur in joint replacements, fracture management, and bone grafting procedures. Rapid and accurate pathogen-specific diagnostic methods are urgently needed to support early clinical decisions. Current culture-based methods are slow and delay effective treatment. This study evaluated the diagnostic value of combining Raman microscopy with high-resolution micro-computed tomography (micro-CT). Human bone samples, either uninfected or inoculated with S. aureus or S. epidermidis, were analyzed. Raman spectroscopy detected distinct spectral changes in inoculated bones, including reduced intensity of phosphate (v₁PO₄³⁻), Amide III, and CH₂ deformation bands. A single principal component explained 96-98% of the variance in these infection-related markers. Specifically, the v₁PO₄³⁻ and CH₂ deformation bands effectively differentiated between S. aureus and S. epidermidis infections, capturing 99-100% variance. Micro-CT analysis showed significant structural changes in inoculated bones. Trabecular volume, number, and spacing were particularly affected. Among these, VOX-BV/TV and Mean1 best differentiated between S. aureus and S. epidermidis infections (both p < 0.0001). Support vector machine (SVM) classification repeated stratified k-folg cross-validation accurately detected inoculation status. Combining Raman and micro-CT features yielded moderately improved classification performance in pathogen-specific discrimination. These findings demonstrate that combining molecular (Raman spectroscopy) and structural (micro-CT) methods allows rapid, nondestructive diagnosis of bone infections. This multimodal approach may improve diagnostic precision, supports timely clinical decisions, and ultimately improves patient outcomes in orthopedic and trauma surgery.

Keywords: bone quality, RAMAN microscopy, Micro-computed tomography, principal component analyses, Staphylococcus aureus, Staphylococcus epidermidis, bone infection diagnostics, Bone graft

Received: 24 Apr 2025; Accepted: 04 Aug 2025.

Copyright: © 2025 Lindtner, Kampik, Noack, Putzer, Arora, Coraça-Huber, Schirmer, Degenhart, Lackner, Popp, Badzoka, Kappacher, Huck and Pallua. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Johannes Dominikus Pallua, Innsbruck Medical University, Innsbruck, 6020, Tyrol, Austria

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