Hepatic cirrhosis and recovery as reflected by Raman spectroscopy: information revealed by statistical analysis might lead to a prognostic biomarker.
Hepatic cirrhosis is a severe chronic disease of the liver accompanied by massive changes in the physiology of the cells constituting the hepatic tissue. Success or failing of a therapeutic effort is difficult to recognize because of its late manifestation in the tissue morphology. In this study, the complex course of hepatic cirrhosis and its regression is followed in a rodent carbon tetrachloride model. Raman spectroscopy, which senses molecular vibrations and reflects the molecular composition of a sample, was applied to gain label- and destruction-free insights into the process of cirrhosis and to conclude on the hepatic disease state on the cellular level. Hematoxylin and eosin staining and immunofluorescence labeling were used to obtain complementary information. False color images derived from maps of Raman spectra by spectral unmixing revealed individual nucleus positions giving structural information. Spectral data unraveled chemical changes associated with liver damage on the cellular level. Upon carbon tetrachloride treatment, a higher lipid content and the presence of catabolic products indicated cirrhosis in tissue samples. Furthermore, 77 % of the Raman spectra recorded from treated rats were classified as diseased, whereas 96 % of the Raman spectra recorded from untreated rats were classified as healthy. Importantly, samples from rats that experienced a recovery period revealed a chemical composition highly similar to the ones from healthy rats while morphologically clear signs of tissue damage were still obvious. Eighty-three percent of their Raman spectra were classified as healthy. The vibrational fingerprint of tissue provides characteristic information that might serve as prognostic biomarker. Graphical Abstract Images of hematoxylin and eosin stained tissue slices together with an average Raman spectrum belonging to healthy and cirrhotic rat liver, respectively, visualize morphological and spectral differences between the two states of the liver.
Leibniz Institute of Photonic Technology, Albert-Einstein-Str. 9, 07745, Jena, Germany.|Center for Sepsis Control and Care, Jena University Hospital, Am Klinikum 1, Jena, Germany.
Kerstin
K
Galler
Center for Sepsis Control and Care, Jena University Hospital, Am Klinikum 1, Jena, Germany.|Department of Anesthesiology and Intensive Care Medicine, Jena University Hospital, Erlanger Allee 101, 07747, Jena, Germany.
Esther
E
Fröhlich
Center for Sepsis Control and Care, Jena University Hospital, Am Klinikum 1, Jena, Germany.|Department of Anesthesiology and Intensive Care Medicine, Jena University Hospital, Erlanger Allee 101, 07747, Jena, Germany.
Andreas
A
Kortgen
Center for Sepsis Control and Care, Jena University Hospital, Am Klinikum 1, Jena, Germany.|Department of Anesthesiology and Intensive Care Medicine, Jena University Hospital, Erlanger Allee 101, 07747, Jena, Germany.|InfectoGnostics Forschungscampus Jena, Center for Applied Research, Philosophenweg 7, 07743, Jena, Germany.
Michael
M
Bauer
Leibniz Institute of Photonic Technology, Albert-Einstein-Str. 9, 07745, Jena, Germany.|Center for Sepsis Control and Care, Jena University Hospital, Am Klinikum 1, Jena, Germany.|InfectoGnostics Forschungscampus Jena, Center for Applied Research, Philosophenweg 7, 07743, Jena, Germany.|Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany.
Jürgen
J
Popp
Leibniz Institute of Photonic Technology, Albert-Einstein-Str. 9, 07745, Jena, Germany. ute.neugebauer@med.uni-jena.de.|Center for Sepsis Control and Care, Jena University Hospital, Am Klinikum 1, Jena, Germany. ute.neugebauer@med.uni-jena.de.|InfectoGnostics Forschungscampus Jena, Center for Applied Research, Philosophenweg 7, 07743, Jena, Germany. ute.neugebauer@med.uni-jena.de.|Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany. ute.neugebauer@med.uni-jena.de.
Ute
U
Neugebauer
D000818:Animals; D015415:Biomarkers / Q000032:analysis*; D002251:Carbon Tetrachloride; D003627:Data Interpretation, Statistical; D008099:Liver / Q000000981:diagnostic imaging*; D008106:Liver Cirrhosis, Experimental / Q000000981:diagnostic imaging*; D008297:Male; D011379:Prognosis; D017208:Rats, Wistar; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013194:Staining and Labeling
D015415:Biomarkers; D002251:Carbon Tetrachloride
Cirrhosis; Liver; N-FINDR; PCA-LDA; Raman spectroscopy; Spectral histopathology