Artikel
Raman spectroscopy reveals inflammation-related changes of lipids following spinal cord injury
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Autoren
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Sandra Tamosaityte
- Klinisches Sensoring und Monitoring - Klinik für Anästhesiologie und Intensivtherapie, TU Dresden, Dresden, Germany
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Roberta Galli
- Klinisches Sensoring und Monitoring - Klinik für Anästhesiologie und Intensivtherapie, TU Dresden, Dresden, Germany
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Ortrud Uckermann
- Neurochirurgie, Universitätsklinikum Carl Gustav Carus, TU Dresden, Dresden, Germany
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Kerim H. Sitoci-Ficici
- Neurochirurgie, Universitätsklinikum Carl Gustav Carus, TU Dresden, Dresden, Germany
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Gerald Steiner
- Klinisches Sensoring und Monitoring - Klinik für Anästhesiologie und Intensivtherapie, TU Dresden, Dresden, Germany
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Matthias Kirsch
- Neurochirurgie, Universitätsklinikum Carl Gustav Carus, TU Dresden, Dresden, Germany
| Veröffentlicht: | 8. Juni 2016 |
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Gliederung
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Objective: Spinal cord injury (SCI) induces heterogeneous alterations of nervous tissue lipids. The demyelination of axons as well as consecutive presence of lipid-laden activated microglia and macrophages, which remove myelin derived lipid debris and thus often transform into foam cells, take place. Using an experimental rat model of SCI, Raman micro-spectroscopy was applied and confirmed by Coherent anti-Stokes Raman scattering (CARS) and two-photon excited fluorescence (TPEF) microscopy to assess the morphochemical information about alterations of lipid distribution following SCI.
Method: Longitudinal cryosections of Wistar rats spinal cords were prepared 21 days after hemisection-induced injury at the level T9 of thoracic vertebrae. Five animals were included into study. The unstained sections on CaF2 slides and consecutive sections on glass slides were prepared for Raman spectroscopy as well as CARS/TPEF microscopy and H&E histology, respectively.
Results: Biochemical information about inflammation-related alterations of lipids was retrieved by addressing specific Raman spectral features. (i) The injury core was depleted from lipids as showed the intensity maps of Raman lipid-related spectral band at 1440 cm-1. (ii) Preserved white matter and inflammatory regions were characterized by myelin fragmentation and the presence of foam cells which were identified by mapping esterified lipids using the Raman specific spectral band at 1743 cm-1. These findings were in agreement with CARS/TPEF microscopy. (iii) By applying pricipal principal component analysis to the Raman data inflammatory regions were found to be rich in saturated fatty acids, which likely play a role in the lipid metabolism of activated microglia/macrophages.
Conclusions: Raman spectroscopy proved to be a useful technique to specifically address inflammation after SCI and myelin degradation products. Raman spectroscopy is non-invasive and can be applied in vivo, providing a new and valuable tool for the diagnosis of the tissue status and monitoring of regenerative therapies.
