Detection of microvasculature in rat hind limb using synchrotron radiation.

BACKGROUND New X-ray microangiography and third-generation synchrotron radiation-based micro-computed tomography have opened new perspectives for microvascular imaging of extremity. Here we aimed to visualize deep-level microvascular structure in rat hind limb by microangiographic technique, and compare images with those by conventional method. MATERIALS AND METHODS A total of 10 Sprague Dawley rats were used for in vivo and ex vivo study (five rats/group). Microangiography in vivo and ex vivo was performed and images were compared with those by conventional method. Synchrotron radiation-based micro-computed tomography (SRμCT) was also performed to reveal three-dimensional (3D) morphology of the blood vessel in rat hind limb. RESULTS By microangiographic technique, blood vessels in the rat limb could be clearly depicted with the minimum visualized blood vessel about 9 μm in diameter, and higher angiographic scores were achieved than those by conventional X-ray. In addition, the vascular network could be defined and analyzed at the micrometer scale from the 3D renderings of limb vessel as shown by SRμCT. CONCLUSIONS Synchrotron radiation-based microangiography and SRμCT thus provided a practical and effective means to observe the microvasculature of limbs, which might be useful in assessment of angiogenesis in lower limbs.

[1]  Dilation of perforating arteries in rat brain in response to systemic hypotension is more sensitive and pronounced than that of pial arterioles: simultaneous visualization of perforating and cortical vessels by in-vivo microangiography. , 2009, Microvascular research.

[2]  Atsushi Momose,et al.  Phase–contrast X–ray computed tomography for observing biological soft tissues , 1996, Nature Medicine.

[3]  W. Pevec,et al.  New blood vessels can be induced to invade ischemic skeletal muscle. , 1996, Journal of vascular surgery.

[4]  Laurent Risser,et al.  From Homogeneous to Fractal Normal and Tumorous Microvascular Networks in the Brain , 2007, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[5]  Bert Müller,et al.  Blood vessel staining in the myocardium for 3D visualization down to the smallest capillaries , 2006 .

[6]  M Ochiai,et al.  Use of synchrotron radiation microangiography to assess development of small collateral arteries in a rat model of hindlimb ischemia. , 1997, Circulation.

[7]  Hidezo Mori,et al.  Visualization of Penetrating Transmural Arteries In Situ by Monochromatic Synchrotron Radiation , 1994, Circulation.

[8]  K. Sugimura,et al.  Synchrotron Radiation Coronary Microangiography for Morphometric and Physiological Evaluation of Myocardial Neovascularization Induced by Endothelial Progenitor Cell Transplantation , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[9]  T. Aach,et al.  A technique to detect and to quantify fasciocutaneous blood vessels in small laboratory animals ex vivo. , 2006, The Journal of surgical research.

[10]  WeimingLi,et al.  High-Resolution Quantitative Computed Tomography Demonstrating Selective Enhancement of Medium-Size Collaterals by Placental Growth Factor-1 in the Mouse Ischemic Hindlimb , 2006 .

[11]  H. Riesemeier,et al.  Going beyond histology. Synchrotron micro-computed tomography as a methodology for biological tissue characterization: from tissue morphology to individual cells , 2009, Journal of The Royal Society Interface.

[12]  J. Isner,et al.  Direct intramuscular gene transfer of naked DNA encoding vascular endothelial growth factor augments collateral development and tissue perfusion. , 1996, Circulation.

[13]  K. Kangawa,et al.  Imaging of the pulmonary circulation in the closed-chest rat using synchrotron radiation microangiography. , 2007, Journal of applied physiology.

[14]  K. Hyodo,et al.  Sex difference in peripheral arterial response to cold exposure. , 2008, Circulation journal : official journal of the Japanese Circulation Society.

[15]  Tao-Sheng Li,et al.  Angiogenesis induced by the injection of peripheral leukocytes and platelets. , 2002, The Journal of surgical research.

[16]  H. Hosaka,et al.  Small-vessel radiography in situ with monochromatic synchrotron radiation. , 1996, Radiology.

[17]  SatoshiTakeshita,et al.  Endothelium-Dependent Relaxation of Collateral Microvessels After Intramuscular Gene Transfer of Vascular Endothelial Growth Factor in a Rat Model of Hindlimb Ischemia , 1998 .

[18]  E. Brogi,et al.  Therapeutic angiogenesis. A single intraarterial bolus of vascular endothelial growth factor augments revascularization in a rabbit ischemic hind limb model. , 1994, The Journal of clinical investigation.

[19]  P Cloetens,et al.  X‐ray high‐resolution vascular network imaging , 2004, Journal of microscopy.

[20]  F. Berthiaume,et al.  Isolated Perfusion of a Tubed Superficial Epigastric Flap in a Rodent Model: 44. , 2006 .

[21]  Atsushi Momose,et al.  Vessel Imaging by Interferometric Phase-Contrast X-Ray Technique , 2002, Circulation.

[22]  Imo E. Hoefer,et al.  Role of Ischemia and of Hypoxia-Inducible Genes in Arteriogenesis After Femoral Artery Occlusion in the Rabbit , 2001, Circulation research.

[23]  T. Yamashita Evaluation of the Microangioarchitecture of Tumors by Use of Monochromatic X-Rays , 2001, Investigative radiology.

[24]  Philipp Schneider,et al.  Hierarchical microimaging for multiscale analysis of large vascular networks , 2006, NeuroImage.

[25]  K. Hyodo,et al.  Cigarette-smoke-induced vasoconstriction of peripheral arteries: evaluation by synchrotron radiation microangiography. , 2007, Circulation journal : official journal of the Japanese Circulation Society.