A Novel and Sensitive Approach for the Evaluation of Liver Ischemia-Reperfusion Injury After Liver Transplantation

ObjectivesThe purpose of our study was to evaluate the potential of x-ray propagation-based phase-contrast imaging (PCI) computed tomography (CT) for the detection and characterization of early changes after ischemia-reperfusion (IR) in a standardized rat liver transplantation (LTx) model. Materials and MethodsSyngeneic orthotopic liver transplantation was performed in male Lewis rats. Ischemia-reperfusion injury (IRI)–induced changes of liver parenchyma were investigated in a time-dependent manner (2, 16, 24, and 32 hours). X-ray phase-contrast images of formalin-fixated liver specimens were acquired in CT mode by using a voxel size of 8 × 8 × 8 &mgr;m3. Necrapoptotic cell death was visualized with the TdT-mediated dUTP-biotin nick end labeling technique, and alterations of liver graft microhemodynamics, that is, acinar and sinusoidal perfusion failure, were evaluated by in vivo fluorescence microscopy. ResultsAcquired and reconstructed PCI-CT images showed an increase in necrotic liver parenchyma dependent on cold storage time, measuring 5.7% ± 1.6% after 2 hours (comparable to 2.6% ± 0.4% for sham livers), 11.5% ± 2.1% (16 hours; P < 0.05 vs control), 23.0% ± 0.5% (24 hours; P < 0.001 vs control), and 31.3% ± 2.2% (32 hours; P < 0.001 vs control). There were a significant lower number of perfused acini in dependence on increasing cold storage time. The acinar perfusion index reached 0.970 ± 0.006 after 2 hours of cold ischemia (comparable to 0.960 ± 0.009 for sham livers) and declined continuously after 16, 24, and 32 hours cold ischemia (0.58 ± 0.03, 0.49 ± 0.02, 0.41 ± 0.03, each P < 0.0001 vs controls). Comparable results were found for sinusoidal perfusion, reaching 1.8% ± 0.4% of nonperfused sinusoids for 2 hours of cold ischemia and 8.2% ± 0.8% after 16 hours, 18.8% ± 1.4% after 24 hours, and 39.0% ± 2.4% after 32 hours (each P < 0.0001 vs controls). Prolonged cold ischemia was associated with an increasing number of TdT-mediated dUTP-biotin nick end labeling-positive cells (hepatocytes and sinusoidal lining cells), reaching 0.4 ± 0.1 (sham), 0.7 ± 0.4 (2 hours), 6.4 ± 1.1 (16 hours), 2.1 ± 0.3 (24 hours), and 14.7 ± 3.5 (32 hours; P = 0.002) for hepatocytes. ConclusionsX-ray PCI of histological liver specimens can detect IR-induced tissue necrosis and can provide detailed complementary 3-dimensional information to standard histopathologic findings.

[1]  E Brun,et al.  High-resolution breast tomography at high energy: a feasibility study of phase contrast imaging on a whole breast , 2012, Physics in medicine and biology.

[2]  L. Pączek,et al.  [Liver transplantation for hepatocellular carcinoma]. , 2003, Polskie Archiwum Medycyny Wewnetrznej.

[3]  Paola Coan,et al.  X-ray phase-contrast imaging: from pre-clinical applications towards clinics , 2013, Physics in medicine and biology.

[4]  W. Peng,et al.  Visualising liver fibrosis by phase-contrast X-ray imaging in common bile duct ligated mice , 2012, European Radiology.

[5]  Zdenka Kuncic,et al.  Image quality in thoracic 4D cone-beam CT: a sensitivity analysis of respiratory signal, binning method, reconstruction algorithm, and projection angular spacing. , 2014, Medical physics.

[6]  M. Menger,et al.  Differential impact of carolina rinse and university of wisconsin solutions on microcirculation, leukocyte adhesion, kupffer cell activity and biliary excretion after liver transplantation , 1993, Hepatology.

[7]  C. Zuiani,et al.  Post-operative imaging in liver transplantation: state-of-the-art and future perspectives. , 2014, World journal of gastroenterology.

[8]  H. Schlitt,et al.  Adenoviral bcl-2 Transfer Improves Survival and Early Graft Function after Ischemia and Reperfusion in Rat Liver Transplantation , 2005, Transplantation.

[9]  Emmanuel Brun,et al.  High-resolution, low-dose phase contrast X-ray tomography for 3D diagnosis of human breast cancers , 2012, Proceedings of the National Academy of Sciences.

[10]  M. Mendenhall,et al.  Pulsed tunable monochromatic X-ray beams from a compact source: new opportunities. , 2003, AJR. American journal of roentgenology.

[11]  S. Yagi,et al.  Comprehensive and innovative techniques for liver transplantation in rats: a surgical guide. , 2010, World journal of gastroenterology.

[12]  F. Pfeiffer,et al.  Imaging Liver Lesions Using Grating-Based Phase-Contrast Computed Tomography with Bi-Lateral Filter Post-Processing , 2014, PloS one.

[13]  Jian Jun Hu,et al.  The monitoring of microvascular liver blood flow changes during ischemia and reperfusion using laser speckle contrast imaging. , 2014, Microvascular research.

[14]  K. Messmer,et al.  Platelet-endothelial cell interactions during hepatic ischemia-reperfusion in vivo: a systematic analysis. , 2003, Microvascular research.

[15]  Franz Pfeiffer,et al.  Quantitative X-ray phase-contrast computed tomography at 82 keV. , 2013, Optics express.

[16]  Pierre Soille,et al.  Morphological image compositing , 2006, IEEE Transactions on Pattern Analysis and Machine Intelligence.

[17]  A. Yoneyama,et al.  Enhanced renal image contrast by ethanol fixation in phase-contrast X-ray computed tomography. , 2014, Journal of synchrotron radiation.

[18]  R. Ruth,et al.  X-ray phase-contrast tomography with a compact laser-driven synchrotron source , 2015, Proceedings of the National Academy of Sciences.

[19]  Franz Pfeiffer,et al.  Multimodal imaging of human cerebellum - merging X-ray phase microtomography, magnetic resonance microscopy and histology , 2012, Scientific Reports.

[20]  H. Jaeschke,et al.  Apoptosis versus oncotic necrosis in hepatic ischemia/reperfusion injury. , 2003, Gastroenterology.

[21]  V. Kupčová,et al.  [Indications and contraindications for liver transplantation]. , 1996, Bratislavske lekarske listy.

[22]  K. Jauch,et al.  Impact of prolonged cold ischemia and reperfusion on apoptosis, activation of caspase 3, and expression of bax after liver transplantation in the rat. , 2001, Transplantation proceedings.

[23]  H. O. Wyckoff,et al.  The International Commission on Radiation Units and Measurements , 2001, Journal of the ICRU.

[24]  Yi-Cyun Yang,et al.  Molecular imaging of ischemia and reperfusion in vivo with mitochondrial autofluorescence. , 2014, Analytical chemistry.

[25]  S. Wilkins,et al.  Simultaneous phase and amplitude extraction from a single defocused image of a homogeneous object , 2002, Journal of microscopy.

[26]  H. Rusinek,et al.  Advanced liver fibrosis: diagnosis with 3D whole-liver perfusion MR imaging--initial experience. , 2008, Radiology.

[27]  E. Hui,et al.  In vivo DTI assessment of hepatic ischemia reperfusion injury in an experimental rat model , 2009, Journal of magnetic resonance imaging : JMRI.

[28]  M. Stegall,et al.  Risk factors for primary dysfunction after liver transplantation--a multivariate analysis. , 1993, Transplantation.

[29]  M M Murnane,et al.  Bright, coherent, ultrafast soft X-ray harmonics spanning the water window from a tabletop light source. , 2010, Physical review letters.

[30]  Emmanuel Brun,et al.  A Dictionary Learning Approach with Overlap for the Low Dose Computed Tomography Reconstruction and Its Vectorial Application to Differential Phase Tomography , 2014, PloS one.

[31]  K. Messmer,et al.  THE IMPACT OF ARTERIALIZATION ON HEPATIC MICROCIRCULATION AND LEUKOCYTE ACCUMULATION AFTER LIVER TRANSPLANTATION IN THE RAT , 1992, Transplantation.

[32]  S. Shen,et al.  Determination of apparent diffusion coefficient to quantitatively study partial hepatic ischemia reperfusion injury in a rabbit model. , 2011, Transplantation Proceedings.

[33]  M. Salvadori,et al.  What's new in clinical solid organ transplantation by 2013. , 2014, World journal of transplantation.

[34]  R. Calne,et al.  ORTHOTOPIC LIVER TRANSPLANTATION IN THE RAT: TECHNIQUE USING CUFF FOR PORTAL VEIN ANASTOMOSIS AND BILIARY DRAINAGE , 1979, Transplantation.