Detection Of Angiogenesis In Hepatocellular Carcinoma With Synchrotron Radiation

Part1Hepatocellular Carcinoma and Angiogenesis Imaging Using Synchrotron Radiation ex vivoObjective To investigate the murine hepatocellular carcinoma angiogenesis ex vivo using microangiography with synchrotron radiation.Methods HCCLM3-RFP cells were seeded into mice liver to establish xenograft mouse models. On the7th,14th,21th and28th day after the implantation,3male HCCLM3-RFP mice were anaesthetized by injection of pentobarbital sodium in lower abdominal wall, respectively. Saline was infused via inferior vena cave through an intracatheter by hand pressure until all the blood in the liver was displaced. On the28th day, another3male HCCLM3-RFP mice were anaesthetized, and barium sulfate was infused until the vessels in the liver and tumor appeared white. After infusion, arteries, veins and common bile duct were ligated, and the livers were excised and fixed with formalin for imaging. Experiments were performed at the beamline BL13W of Shanghai Synchrotron Radiation Facility (SSRF) in China.Results Combining the CT technique, high resolution images of tumor angiogenesis were acquired. Normal blood vessels appear to be displaced by the fast growing tumors. Abundant and tortuous tumor angiogenesis in the tumor periphery area and sparse angiogenesis inside the tumor could be visualized clearly. The smallest tumor vessels visualized were approximately20μm in diameter. Judging from the statistical results of microvessel density, microvessel density reached the maximum, approximately4.58%at day14. After day14, an obvious volume increase was observed and growth rate attain the maximum, but there was a noticeable decline in microvessel density.Conclusions It is demonstrated that microangiography with synchrotron radiation using saline and barium sulfate as contrast agent is a viable imaging strategy to image tumor angiogenesis. Part IIIn-line X-ray phase-contrast imaging of human hepatocellular carcinoma microvasculature ex vivoObjectives To evaluate the microangioarchitecture of human hepatocellular carcinoma (HCC) ex vivo using in-line phase-contrast (IL-PCI) X-ray imaging with synchrotron radiation.Methods From June2011to October2011,5surgical specimens of human HCC were obtained from the Department of Hepatobiliary Surgery, Zhongshan Hospital, Fudan University. Without infusion of any contract agent before the operation, the specimens were excised and fixed with formalin for imaging. Visualizing HCC microvasculature ex vivo with IL-PCI was performed at the beamline BL13W of Shanghai Synchrotron Radiation Facility (SSRF) in China.Results Combining the computed tomography technique, high resolution images of tumor angiogenesis were acquired. The tumor boundary and angiogenesis could be detected clearly, and detection of10μm diameter blood vessels could be achieved. Moreover,3D image reconstruction of HCC angiogenesis was obtained. Structural characteristics of HCC vasculature included spatial heterogeneity and chaotic structure, high vascular tortuosity, extreme heterogeneity of vascular density with areas of low vascular density mixed with regions of high angiogenic activity.Conclusion These results highlights the imaging quality of IL-PCI. It is suggested that IL-PCI can successfully reveal the microangioarchitecture of HCC without using any contrast. Part ⅢDiffraction enhanced imaging of human hepatocellular carcinoma microvasculature ex vivoObjectives To evaluate the microangioarchitecture of human hepatocellular carcinoma (HCC) ex vivo using diffraction enhanced imaging (DEI) with synchrotron radiation.Methods From June2011to October2011,5surgical specimens of human HCC were obtained from the Department of Hepatobiliary Surgery, Zhongshan Hospital, Fudan University. Without infusion of any contract agent before the operation, the specimens were excised and fixed with formalin for imaging. Visualizing HCC microvasculature ex vivo with DEI was performed at the beamline4W1A of Beijing Synchrotron Radiation Facility (BSRF) in China.Results By the DEI technique, high resolution images of tumor angiogenesis were acquired. The tumor boundary, angiogenesis and tumor nests could be detected clearly, and detection of25μm diameter blood vessels could be achieved. Structural characteristics of HCC vasculature included spatial heterogeneity and chaotic structure.Conclusion These results highlights the imaging quality of DEI. It is suggested that DEI can successfully reveal the microangioarchitecture of HCC without using any contrast.