Study on the in vitro and in vivo activation of rat hepatic stellate cells by Raman spectroscopy.

The feasibility of a novel and efficient diagnostic method for liver fibrosis using Raman spectroscopy is studied. Confocal Raman spectroscopy (CRS) is utilized to monitor the molecular changes of hepatic stellate cells (HSCs) in vitro as well as in vivo activation. In vitro activation was induced by growth in uncoated plastic plates, while the in vivo activation is induced by a single intraperitoneal injection of carbon tetrachloride (CCl(4)). The biochemical changes of HSCs during activation such as the loss of retinoid, the increase of alpha-helical protein, and the increased production of extracellular matrix proteins are observed by CRS. A user-friendly autoclassifying system is also developed to classify Raman spectra of liver injury tissues with a 90% accuracy rate. Raman spectroscopy combined with a fiber optical probe could be potentially accomplished for in vivo detection, which can lead to a novel and efficient diagnosis for liver fibrosis.

[1]  B. Wilson,et al.  The Effects of ex vivo Handling Procedures on the Near‐Infrared Raman Spectra of Normal Mammalian Tissues , 1996, Photochemistry and photobiology.

[2]  S. Friedman Molecular Regulation of Hepatic Fibrosis, an Integrated Cellular Response to Tissue Injury* , 2000, The Journal of Biological Chemistry.

[3]  E. Perrier,et al.  Effects of light, oxygen and concentration on vitamin A1 , 2004 .

[4]  Jürgen Popp,et al.  Raman spectroscopic identification of single yeast cells , 2005 .

[5]  Xin Wang,et al.  Screening of gastric carcinoma cells in the human malignant gastric mucosa by confocal Raman microspectroscopy , 2005 .

[6]  Michael S. Feld,et al.  Histological Classification of Raman Spectra of Human Coronary Artery Atherosclerosis Using Principal Component Analysis , 1999 .

[7]  Gerwin J. Puppels,et al.  Monitoring the Penetration Enhancer Dimethyl Sulfoxide in Human Stratum Corneum in Vivo by Confocal Raman Spectroscopy , 2002, Pharmaceutical Research.

[8]  S. Friedman,et al.  Activation of rat hepatic stellate cells leads to loss of glutathion S‐transferases and their enzymatic activity against products of oxidative stress , 1999, Hepatology.

[9]  Y. Paik,et al.  Apoptosis of hepatic stellate cells in carbon tetrachloride induced acute liver injury of the rat: analysis of isolated hepatic stellate cells. , 2003, Journal of hepatology.

[10]  M. Green,et al.  Vitamin A metabolism: new perspectives on absorption, transport, and storage. , 1991, Physiological reviews.

[11]  Gavin Jell,et al.  In situ non‐invasive spectral discrimination between bone cell phenotypes used in tissue engineering , 2004, Journal of cellular biochemistry.

[12]  W. Campbell,et al.  Characterization of normal and malignant human hepatocytes by Raman microspectroscopy. , 1996, Cancer letters.

[13]  S. Friedman,et al.  Liver fibrosis -- from bench to bedside. , 2003, Journal of hepatology.

[14]  Neville Krasner,et al.  Raman spectroscopy of Alzheimer's diseased tissue , 2004, SPIE BiOS.

[15]  L L Hench,et al.  Spectroscopic study of human lung epithelial cells (A549) in culture: living cells versus dead cells. , 2003, Biopolymers.

[16]  U. Hämmerling,et al.  Retro-retinoids in regulated cell growth and death , 1996, The Journal of experimental medicine.

[17]  K. Lindros,et al.  No significant expression of CYP2E1 in rat liver stellate cells. , 1998, Biochemical pharmacology.

[18]  R. Richards-Kortum,et al.  Raman spectroscopy for the detection of cancers and precancers. , 1996, Journal of biomedical optics.

[19]  T. Kitamoto,et al.  Increased 9,13-di-cis-retinoic acid in rat hepatic fibrosis: implication for a potential link between retinoid loss and TGF-beta mediated fibrogenesis in vivo. , 1999, Journal of hepatology.

[20]  B. Lendl,et al.  Multidimensional information on the chemical composition of single bacterial cells by confocal Raman microspectroscopy. , 2000, Analytical chemistry.

[21]  J. Greve,et al.  Studying single living cells and chromosomes by confocal Raman microspectroscopy , 1990, Nature.

[22]  A. Shen,et al.  Raman scattering properties of human pterygium tissue. , 2005, Journal of biomedical optics.

[23]  A. Reuben Ito becomes a star , 2002, Hepatology.

[24]  S. Friedman,et al.  An immortalized rat liver stellate cell line (HSC-T6): a new cell model for the study of retinoid metabolism in vitro. , 2000, Journal of lipid research.

[25]  H. Bruining,et al.  Raman spectroscopic method for identification of clinically relevant microorganisms growing on solid culture medium. , 2000, Analytical chemistry.

[26]  Christoph Krafft,et al.  Mapping of single cells by near infrared Raman microspectroscopy , 2003 .

[27]  Nan Wu,et al.  Design of auto-classifying system and its application in Raman spectroscopy diagnosis of gastric carcinoma , 2003, Proceedings of the 2003 International Conference on Machine Learning and Cybernetics (IEEE Cat. No.03EX693).