Content uploaded by Ryuta Yasuda
Author content
All content in this area was uploaded by Ryuta Yasuda on Aug 10, 2016
Content may be subject to copyright.
A preview of the PDF is not available
A Method for Serial Selective Arterial Catheterization and Digital Subtraction Angiography in Rodents
Abstract and Figures
Imaging is a key element in the study of many rodent models of human diseases. The application of DSA has been limited in these studies in part because of a lack of a method that allows serial intra-arterial examinations to be performed during an extended period of time. It was our intent to develop and test a method for performing sequential arterial catheterizations and DSA in rats.
Using a transfemoral approach, we subjected 12 adult male Harvey rats to 3 sequential DSA examinations during a 6- to 8-week period. At each examination, 2 selective arterial catheterizations and a DSA were performed. Animals were monitored for ill effects, and images from the 3 examinations were compared for quality and the presence of any arterial injury.
Ten of the 12 rats survived all 3 examinations. There were no adverse effects noted and no evidence of arterial injury from the examinations.
With the technique described, it is possible to perform serial arterial catheterizations and DSA in rats. This technique will be useful as an adjunct in the use of rodents for the study of human diseases.
Figures - uploaded by Ryuta Yasuda
Author content
All figure content in this area was uploaded by Ryuta Yasuda
Content may be subject to copyright.
... According to Barth [33], before induction of SAH, a burr hole right side to the midline was made rostrally to the interparietal-occiptal suture and placed a special bulb into cisterna magna, which was connected to a pressure transducer (PowerLab, ADinstrument Co., Australia) through a PE-50 tube. The ICP values of 5 minutes before SAH and 5, 10,15,20,25,30,35,40,45,50,55, 60 minutes after SAH were recorded. ...
... DSA, magnetic resonance angiography (MRA) and CTA are widely used to detect CV in humans. However, the relative low resolution of DSA, MRA and CTA limits the application in small animal models [8,40]. Up to now, most of experimental SAH studies were based on indirect observation or pathological outcomes. ...
Precise in vivo evaluation of cerebral vasospasm caused by subarachnoid hemorrhage has remained a critical but unsolved issue in experimental small animal models. In this study, we used synchrotron radiation angiography to study the vasospasm of anterior circulation arteries in two subarachnoid hemorrhage models in rats. Synchrotron radiation angiography, laser Doppler flowmetry-cerebral blood flow measurement, [(125)I]N-isopropyl-p-iodoamphetamine cerebral blood flow measurement and terminal examinations were applied to evaluate the changes of anterior circulation arteries in two subarachnoid hemorrhage models made by blood injection into cisterna magna and prechiasmatic cistern. Using synchrotron radiation angiography technique, we detected cerebral vasospasm in subarachnoid hemorrhage rats compared to the controls (p<0.05). We also identified two interesting findings: 1) both middle cerebral artery and anterior cerebral artery shrunk the most at day 3 after subarachnoid hemorrhage; 2) the diameter of anterior cerebral artery in the prechiasmatic cistern injection group was smaller than that in the cisterna magna injection group (p<0.05), but not for middle cerebral artery. We concluded that synchrotron radiation angiography provided a novel technique, which could directly evaluate cerebral vasospasm in small animal experimental subarachnoid hemorrhage models. The courses of vasospasm in these two injection models are similar; however, the model produced by prechiasmatic cistern injection is more suitable for study of anterior circulation vasospasm.
... Previous experimental animal models' design forced intra-arterial approaches from the extremities or trunk at the expense of peripheral arteries [7]. However, in our approach, we do not sacrifice any arteries in the extremities or trunk and are able to perform a minimally invasive transarterial approach. ...
Surgery in humans is continuously evolving and promoted minimally invasive treatment. On the other hand, despite the importance of the 3Rs principles for experimental animals is well documented, no reports describe specific methodologies for implementing "refinement" in practice. Here, we describe a new technique, the "Ohta Method" for caudal arthrocentesis in the pursuit of the 3Rs for animal experiments and the development of innovative methods for investigating systemic organ arteries through minimally invasive procedures. This procedure requires only a percutaneous puncture of the caudal artery without any injury to the limb or body trunk. In addition, it does not cut down the artery, making hemostasis easier and recovering arterial damage easier. We will show multiple organ artery angiographies in marmoset for the first time in the world. The principle described in this paper could also be applied to many other small animals, such as rats. Moreover, using this method, multiple doses of the drug or cells can be administered to the target organ at the time of therapeutic intervention, thereby enabling the establishment of more sophisticated and complex therapeutic intervention studies as translational research.
... A transfemoral approach leaves both ICA and ECA intact [12,13]. With the availability of fluoroscopy equipment accessible to small animal laboratories, endovascular approaches using commercially available or custom-made wires used for endovascular procedures have been utilized for tMCAo [3,[12][13][14][15]. Previously, we described the use of a custom-made wire via a femoral artery approach to induce tMCAo [3]. ...
Background
Animal models of stroke play a crucial role in determining the pathophysiology of stroke progression and assessment of any new therapeutic approaches. Transient middle cerebral artery occlusion (tMCAo) in rodent models are the most common site-specific type of ischemia because of their relevance to the clinical setting. Compared with the intraluminal filament technique for inducing tMCAo, the transfemoral approach using endovascular wires is relatively a new technique
Methods
Here we present the use of commercially available wires used for neuro-endovascular surgical procedures to induce tMCAo in rats via a transfemoral approach. We used male Wistar rats in four groups to assess the effect of occlusion time (1 vs. 2 hours) and the wire type (PT2 TM 0.014″ vs. TransendTM EX, 0.014″, Boston Scientific, MA, USA). Infarct volume, edema, neurological deficits, and pro-inflammatory/anti-inflammatory blood biomarkers were used as outcome measures.
Results
We observed a significant effect of the wire type on the infarct volume (p value = 0.0096) where infarcts were slightly larger in the PT2 wiregroups. However, the occlusion time had no significant effect on infarct volume, even though the interaction between wire-type * occlusion-time was significant (p value = 0.024). Also, the amount of edema and blood pro-inflammatory/anti-inflammatory biomarkers were not statistically different among the wire-type and occlusion-time groups.
Conclusions
The choice of appropriate endovascular wire should probably be the focus of the study design instead of the occlusion time when planning an experiment. The transfemoral approach using endovascular wires for inducing tMCAo in rats provides a more consistent outcome with fewer complications compared with suture filament models.
... [3][4][5][6][7][8][9] With the recent advent of angiography procedures, endovascular approaches using commercially available microwires are suggested for temporary MCAo. [10][11][12][13] In this study, we present the technical steps in prototyping custom made microwires with different diameters that allowed for precise occlusion of the MCA under direct fluoroscopic visualization. After determining the optimal microwire diameter/type, experiments were performed in Wistar rats to assess the performance of the designed microwire in inducing MCAo stroke. ...
The aim of this study was to develop a reliable and repeatable method of inducing focal middle cerebral artery occlusion (MCAo) in rats without ligation of the external carotid artery (ECA), while reducing the risk of subarachnoid hemorrhage.
We prototyped microwires with different diameters (0.0120 inch, 0.0115 inch, 0.0110 inch), materials, and construction methods (coil-on-core, extruded polymer jacket-on-core). Under fluoroscopic guidance and using femoral artery access, the microwires were navigated into the internal carotid artery of male Wistar rats (n=50, weight 376±64 g) to induce MCAo for 1 or 2 h. We performed neurological assessments at baseline, and at 3, 24, 72, and 168 h after MCAo. MRI measurements were performed on a 9.4 T scanner at 1 and 7 days post-injury.
The 0.0115 inch microwire with polymer jacket-on-core provided the most successful outcome. At 1 and 7 days post-injury, we observed similar infarction volumes for 1 and 2 h MCAo in the MRI study. Infarcted lesion volumes in both MCAo groups were significantly reduced at 7 days compared with 1 day post-injury. The trend in longitudinal changes for the scores of different neurological assessments was confirmed to be significant after the injury, but both groups showed a similar trend of neurological deficits over the course of the study.
We have developed a reliable and repeatable MCAo method in rats, allowing for precise occlusion of the MCA under direct fluoroscopic visualization without alteration of the cerebral hemodynamics associated with ECA ligation. The custom designed microwire can also be sized for targeted focal ischemia in larger animals.
Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.
... Crucial points for successful DSA of murine cerebral vessels and the detection of vessel caliber changes are 1) a small pixel size (approximately 15 m in our study), 2) a sufficient contrastto-noise ratio, which we achieved by intra-arterial administration of contrast agent, and 3) high temporal resolutions. Since superselective transfemoral catheterization of supraaortic vessels is feasible in rats, 11,27 but not in mice, operative cannulation of the ECA was necessary in our experiments. Thus, the use of a small catheter to fit within the ECA and the process of cannulation of the ECA were other critical steps that required some training. ...
Investigation of the anatomy, patency, and blood flow of arterial and venous vessels in small animal models of cerebral ischemia, venous thrombosis, or vasospasm is of major interest. However, due to their small caliber, in vivo examination of these vessels is technically challenging. Using micro-CT, we compared the feasibility of in vivo DSA and CTA of the murine cerebrovasculature using an intra-arterial route of contrast administration.
The ECA was catheterized in 5 C57BL/6J mice. During intra-arterial injection of an iodized contrast agent (30 μL/1 sec), DSA of the intra- and extracranial vessels was performed in mice breathing room air and repeated in hypoxic/hypercapnic mice. Micro-CTA was performed within 20 seconds of intra-arterial contrast injection (220 μL/20 sec). Image quality of both methods was compared. Radiation dose measurements were performed with thermoluminescence dosimeters.
Both methods provided high-resolution images of the murine cerebrovasculature, with the smallest identifiable vessel calibers of ≤50 μm. Due to its high temporal resolution of 30 fps, DSA allowed identification of anastomoses between the ICA and ECA by detection of retrograde flow within the superficial temporal artery. Micro-CTA during intra-arterial contrast injection resulted in a reduced injection volume and a higher contrast-to-noise ratio (19.0 ± 1.0) compared with DSA (10.0 ± 1.8) or micro-CTA when using an intravenous injection route (1.3 ± 0.4).
DSA of the murine cerebrovasculature is feasible using micro-CT and allows precise and repeated measurements of the vessel caliber, and changes of the vessel caliber, while providing relevant information on blood flow in vivo.
... To meet this need, the technology for in vivo, small animal imaging has advanced rapidly in recent years to include magnetic resonance microscopy(Johnson, Hedlund et al. 1992), micro-CT(Holdsworth and Thornton 2002), micro-ultrasound(Hoit 2001), micro-PET/SPECT(Herschman 2003), and optical methods based on fluorescence and bioluminescence(Ntziachristos, Ripoll et al. 2005). We and others have recently added x-ray-based digital subtraction angiography (DSA) (Buhalog, Yasuda et al. ; Lin, Samei et al. 2006; Badea, Hedlund et al. 2007; Badea, Drangova et al. 2008) to this imaging armamentarium. Drug-induced vascular injury is a particularly challenging drug safety concern for a number of reasons. ...
X-ray based digital subtraction angiography (DSA) is a common clinical imaging method for vascular morphology and function. Coronary artery characterization is one of its most important applications. We show that bi-plane DSA of rat coronary arteries can provide a powerful imaging tool for translational safety assessment in drug discovery.
A novel, dual tube/detector system, constructed explicitly for preclinical imaging, supports image acquisition at 10 frames/s with 88-micron spatial resolution. Ventilation, x-ray exposure, and contrast injection are all precisely synchronized using a biological sequence controller implemented as a LabVIEW application. A set of experiments were performed to test and optimize the sampling and image quality. We applied the DSA imaging protocol to record changes in the visualization of coronaries and myocardial perfusion induced by a vasodilator drug, nitroprusside. The drug was infused into a tail vein catheter using a peristaltic infusion pump at a rate of 0.07 mL/h for 3 min (dose: 0.0875 mg). Multiple DSA sequences were acquired before, during, and up to 25 min after drug infusion. Perfusion maps of the heart were generated in MATLAB to compare the drug effects over time.
The best trade-off between the injection time, pressure, and image quality was achieved at 60 PSI, with the injection of 150 ms occurring early in diastole (60 ms delay) and resulting in the delivery of 113 μL of contrast agent. DSA images clearly show the main branches of the coronary arteries in an intact, beating heart. The drug test demonstrated that DSA can detect relative changes in coronary circulation via perfusion maps.
The methodology for DSA imaging of rat coronary arteries can serve as a template for future translational studies to assist in safety evaluation of new pharmaceuticals. Although x-ray imaging involves radiation, the associated dose (0.4 Gy) is not a major limitation.
Background
The use of digital silhouette angiography (DSA) has been restricted due to lack of a technique that allows for repeated intra-arterial inspections over a prolonged period. Current studies are focused on the arteries that can be cannulated multiple times. We intended to develop and test a technique that would enable repeated performance of various catheterizations and transcatheter operations for a prolonged period, at the same site, with fewer postsurgical complications.
Methods
Thirty rats were randomly divided into five groups. Ventral caudal artery cannulation was performed via the transtail approach after grouping for subsequent experiments. Histological staining and scanning electron microscopy were used to assess endothelial injuries.
Results
The rats survived post catheterization of ventral caudal artery and establishment of animal models. The average time of ventral caudal artery cannulation was significantly shorter than that of the femoral (p < 0.01) and common carotid arteries (p < 0.01). In rats, the transtail artery technique effectively allowed selective arterial catheterization and angiography. Histological staining and scanning electron microscopy of the abdominal aorta revealed disruption of the intima and denuded wavy endothelial surface.
Conclusions
We describe a novel method for artery sheath catheterization through the ventral caudal artery in rats; it may be possible to perform serial DSA studies and interventional operations with a single sheath channel in rats over a prolonged period. We believe that this approach will improve the utility of rats as models of human diseases and enable the broader use of rodent models for endovascular therapy research.
Transtail artery approach successfully enables selective arterial catheterization and angiography in the rat. This technique is effective and repeatable. In addition to its utility in imaging, it may also have a wide range of applications in transcatheter therapy and experimental cerebral stroke models.
Serial magnetic resonance imaging (MRI) was performed to investigate the temporal and spatial relationship between the biphasic nature of blood-brain barrier (BBB) opening and, in parallel, edema formation after ischemia-reperfusion (I/R) injury in rats. T(2)-weighted imaging combined with T(2)-relaxometry, mainly for edema assessment, was performed at 1 h after ischemia, after reperfusion, and at 4, 24 and 48 h after reperfusion. T(1)-weighted imaging was performed before and after gadolinium contrast at the last three time points to assess BBB integrity. The biphasic course of BBB opening with a significant reduction in BBB permeability at 24 h after reperfusion, associated with a progressive expansion of leaky BBB volume, was accompanied by a peak ipsilateral edema formation. In addition, at 4 h after reperfusion, edema formation could also be detected at the contralateral striatum as determined by the elevated T(2)-values that persisted to varying degrees, indicative of widespread effects of I/R injury. The observations of this study may indicate a dynamic temporal shift in the mechanisms responsible for biphasic BBB permeability changes, with complex relations to edema formation. Stroke therapy aimed at vasogenic edema and drug delivery for neuroprotection may also be guided according to the functional status of the BBB, and these findings have to be confirmed in human stroke.
Small-animal imaging has a critical role in phenotyping, drug discovery and in providing a basic understanding of mechanisms of disease. Translating imaging methods from humans to small animals is not an easy task. The purpose of this work is to review in vivo x-ray based small-animal imaging, with a focus on in vivo micro-computed tomography (micro-CT) and digital subtraction angiography (DSA). We present the principles, technologies, image quality parameters and types of applications. We show that both methods can be used not only to provide morphological, but also functional information, such as cardiac function estimation or perfusion. Compared to other modalities, x-ray based imaging is usually regarded as being able to provide higher throughput at lower cost and adequate resolution. The limitations are usually associated with the relatively poor contrast mechanisms and potential radiation damage due to ionizing radiation, although the use of contrast agents and careful design of studies can address these limitations. We hope that the information will effectively address how x-ray based imaging can be exploited for successful in vivo preclinical imaging.
The use of preclinical rodent models of disease continues to grow because these models help elucidate pathogenic mechanisms and provide robust test beds for drug development. Among the major anatomic and physiologic indicators of disease progression and genetic or drug modification of responses are measurements of blood vessel caliber and flow. Moreover, cardiopulmonary blood flow is a critical indicator of gas exchange. Current methods of measuring cardiopulmonary blood flow suffer from some or all of the following limitations--they produce relative values, are limited to global measurements, do not provide vasculature visualization, are not able to measure acute changes, are invasive, or require euthanasia.
In this study, high-spatial and high-temporal resolution x-ray digital subtraction angiography (DSA) was used to obtain vasculature visualization, quantitative blood flow in absolute metrics (ml/min instead of arbitrary units or velocity), and relative blood volume dynamics from discrete regions of interest on a pixel-by-pixel basis (100 x 100 microm2).
A series of calibrations linked the DSA flow measurements to standard physiological measurement using thermodilution and Fick's method for cardiac output (CO), which in eight anesthetized Fischer-344 rats was found to be 37.0 +/- 5.1 ml/min. Phantom experiments were conducted to calibrate the radiographic density to vessel thickness, allowing a link of DSA cardiac output measurements to cardiopulmonary blood flow measurements in discrete regions of interest. The scaling factor linking relative DSA cardiac output measurements to the Fick's absolute measurements was found to be 18.90 x CODSA = COFick.
This calibrated DSA approach allows repeated simultaneous visualization of vasculature and measurement of blood flow dynamics on a regional level in the living rat.
Long-term patency rates of small-diameter expanded polytetrafluoroethylene (ePTFE) vascular prostheses are unsatisfactory. Treatment of ePTFE grafts by alcohol before implantation was reported to increase hydrophilic properties, yielding better endothelialization and cellular in-growth, thus improving graft healing. The effect of alcohol pretreatment on ePTFE grafts and postoperative healing characteristics of wet ePTFE grafts were evaluated in this study. Ten sterile ePTFE grafts (2 mm ID, 30 micro thru-pore, 12 mm long) were implanted in the infrarenal aorta of male Sprague-Dawley rats (324-380 g). Five grafts were treated with ethanol 70% and soaked with saline solution before implantation (wet); five nontreated grafts served as control. All rats were sacrificed after digital subtraction angiography and sampling of the graft for histological investigation after 3 weeks. Histomorphometric analysis was performed for endothelial coverage, cellular in-growth, and intimal hyperplasia. All grafts were patent at the end of 3 weeks in both groups. Histological evaluation revealed significantly better endothelial coverage and prominent infiltration by fibroblasts and lymphocytes in the wet group. Endothelial coverage (31.03 +/- 10.61% vs. 13.03 +/- 9.46%, P = 0.03) and cellular infiltration of grafts (50.91 +/- 8.55% vs. 39.29 +/- 10.70%, P = 0.11) were higher in the wet group. Area of intimal hyperplasia per graft length was also higher in the wet group (5.32 +/- 4.75 microm(2)/microm vs. 2.69 +/- 3.41 microm(2)/microm, P = 0.36). Wetting of ePTFE grafts with ethanol 70% pretreatment before implantation might have a beneficial effect on long-term patency of small-diameter vascular grafts due to facilitated graft healing.
Heparan sulfate proteoglycans (HSPGs) are potent regulators of vascular remodeling and repair. Heparanase is the major enzyme capable of degrading heparan sulfate in mammalian cells. Here we examined the role of heparanase in controlling arterial structure, mechanics, and remodeling. In vitro studies supported that heparanase expression in endothelial cells serves as a negative regulator of endothelial inhibition of vascular smooth muscle cell (vSMC) proliferation. Arterial structure and remodeling to injury were also modified by heparanase expression. Transgenic mice overexpressing heparanase had increased arterial thickness, cellular density, and mechanical compliance. Endovascular stenting studies in Zucker rats demonstrated increased heparanase expression in the neointima of obese, hyperlipidemic rats in comparison to lean rats. The extent of heparanase expression within the neointima strongly correlated with the neointimal thickness following injury. To test the effects of heparanase overexpression on arterial repair, we developed a novel murine model of stent injury using small diameter self-expanding stents. Using this model, we found that increased neointimal formation and macrophage recruitment occurs in transgenic mice overexpressing heparanase. Taken together, these results support a role for heparanase in the regulation of arterial structure, mechanics, and repair.
Long-term patency of conventional synthetic grafts is unsatisfactory below a 6-mm internal diameter. Poly(epsilon-caprolactone) (PCL) is a promising biodegradable polymer with a longer degradation time. We aimed to evaluate in vivo healing and degradation characteristics of small-diameter vascular grafts made of PCL nanofibers compared with expanded polytetrafluoroethylene (ePTFE) grafts.
We prepared 2-mm-internal diameter grafts by electrospinning using PCL (M(n)=80, 000 g/mol). Either PCL (n=15) or ePTFE (n=15) grafts were implanted into 30 rats. Rats were followed up for 24 weeks. At the conclusion of the follow-up period, patency and structural integrity were evaluated by digital subtraction angiography. The abdominal aorta, including the graft, was harvested and investigated under light microscopy. Endothelial coverage, neointima formation, and transmural cellular ingrowth were measured by computed histomorphometry. All animals survived until the end of follow-up, and all grafts were patent in both groups. Digital subtraction angiography revealed no stenosis in the PCL group but stenotic lesions in 1 graft at 18 weeks (40%) and in another graft at 24 weeks (50%) in the ePTFE group. None of the grafts showed aneurysmal dilatation. Endothelial coverage was significantly better in the PCL group. Neointimal formation was comparable between the 2 groups. Macrophage and fibroblast ingrowth with extracellular matrix formation and neoangiogenesis were better in the PCL group. After 12 weeks, foci of chondroid metaplasia located in the neointima of PCL grafts were observed in all samples.
Small-diameter PCL grafts represent a promising alternative for the future because of their better healing characteristics compared with ePTFE grafts. Faster endothelialization and extracellular matrix formation, accompanied by degradation of graft fibers, seem to be the major advantages. Further evaluation of degradation and graft healing characteristics may potentially lead to the clinical use of such grafts for revascularization procedures.
The organ distribution of radiopharmaceuticals in the rat is usually estimated using 7% of body weight (BW) for blood volume (BV). In spite of its important impact on the evaluation of new agents, this value has not been validated adequately. We therefore studied blood volume in 70 awake Wistar rats (100 to 400 g BW) in which red blood cell volume (RBCV) and plasma volume (PV) were measured simultaneously. Red blood cell volume was measured by in vitro RBC-tagging with Tc-99m in Sn-pyrophosphate, 0.05 microgram per ml of blood; plasma volume was measured with I-125 human serum albumin (HSA). Ten minutes after injection of the dose, 0.5 ml of blood was withdrawn from the carotid or femoral artery and duplicate samples of 0.025 ml of blood were counted after separating RBCs from plasma. Total blood volume was calculated by adding RBC volume and plasma volume. The relationship for the entire group was: BV (ml) = 0.06 X BW + 0.77 (r = 0.99, n = 70, p less than 0.001). The difference between male and female rats was not statistically significant. The use of an arbitrary value of 7% for estimation of blood volume can lead to significant errors in calculating radiopharmaceutical distribution. The use of the general formula for the blood-volume calculation described here should improve the accuracy and reliability of estimates of radiopharmaceutical distribution.
To the Editor:
The major disadvantage in rat models of global cerebral ischemia1 2 3 is the difficulty of occluding the vertebral artery through the alar foramina because the electrocoagulations are done “blind.” This mode of vertebral vessel occlusion is often unsatisfactory, and while postoperative selection on the basis of righting responses, EEG, and pupillary size is used in cerebral 4-vessel occlusion models,1 2 3 there is no substitute for demonstrating the completeness of the vascular occlusion . This “blind” vertebral occlusion technique might also explain the wide variation in cerebral blood flow measurements after brain 4-vessel occlusion.4 5 Furthermore, in the case of cerebral 2-vessel …
The aim of this study was to visualize hepatic microvessels (less than 100 microm in diameter) in vivo, which could not be visualized by conventional X-ray angiography, by using synchrotron radiation (ultra-bright and monochromatic X-ray).
Five female Balb/c nu/nu mice were used. To investigate the hepatic microvessels under nearly physiologic conditions, we performed in vivo aortography under anesthesia with 370 mgI/ml nonionic iodine contrast medium using monochromatic 17-keV X-rays generated by a synchrotron. Images were captured with a pixel matrix size of 1024 x 1024 at a rate of 30 pictures/s. The field of view was 7 mm x 7 mm and thus the pixel size was approximately 7 microm. Captured images were evaluated both qualitatively and quantitatively.
Small hepatic arterial and portal venous branches of the liver were visualized separately during one sequential aortogram. The minimum diameter of the vessels observed was approximately 20 microm, and the vessels which ran parallel to the hepatic artery were observed and it seemed to be intrahepatic peribiliary arterial plexus.
Our new experimental model would be useful for visualization of changes in the hepatic microcirculation under nearly physiologic conditions.
Although the rat subarachnoid hemorrhage model is well established in vasospasm research, the angiographic evaluation is difficult due to the animal's small size. For this reason, the aim of the study was to develop a standardized angiographic examination technique without additional complex equipment. Under general anesthesia, 11 Sprague-Dawley rats underwent selective cerebral digital subtraction angiography using a 0.3 mm focal spot and a 2.0-fold linear magnification. Five animals had experimental subarachnoid hemorrhage according to the "double-hemorrhage" model. Comparison with the intraarterial tip of the microcatheter enabled calibration of the vessel lumen. The diameter of the normal basilar artery (n=6) was 0.34+/-0.03 mm (mean+/-SD), whereas delayed vasospastic constriction (mean 6.2 days) caused a reduction in diameter of 32.4% (0.23+/-0.09 mm) as well as impaired collateral blood flow via the posterior communicating artery and anterior spinal artery. Histological examination of sections stained with hematoxylin and eosin under a light microscope confirmed vasospasm. In conclusion, biplane digital subtraction angiography allows precise and reliable evaluation of arterial diameter reduction and hemodynamic parameters in a rat vasospasm model. However, further investigation is required for assessment of vasoactive drugs, e.g., endothelin receptor antagonists.