大鼠颈总动脉端-侧吻合模型的建立

目的建立大鼠颈总动脉端-侧吻合模型,以期对欲进行显微血管吻合训练或相关实验的同道提供帮助。方法成年SD大鼠10只,将左侧颈总动脉远端穿过颈前肌肉群和气管之间的隧道,与右侧颈总动脉行端-侧吻合。结果成功建立大鼠颈总动脉端-侧血管吻合动物模型,手术成功率约为100%,平均吻合所需8针,平均血管吻合所需时间(35±5)min。吻合3个月后观察通畅率100%,HE染色示吻合口愈合良好。结论吻合成功的关键是提高显微操作技术水平,同时注意保护术野中小的血管和神经。此模型可以较好的应用于显微血管缝合训练。     

Comparison of lymphocyte subpopulations in arterial and venous blood of the rat

Lymphocyte subpopulation as well as other hematological properties were compared between arterial and venous bloods of Wistar-Imamichi rats. Lymphocyte subsets were defined with four monoclonal antibodies which were specific to the respective cell surface glycoproteins. Using these monoclonal antibodies, subsets of B lymphocytes, T lymphocytes, helper T lymphocytes, and suppressor and cytotoxic T lymphocytes in the peripheral lymphocytes were identified. The blood samples were taken from aorta abdominalis and venae cava caudalis. The population of these subsets were enumerated by a laser flow-cytometry system. The result showed that there was no significant difference in hematological properties between the arterial and venous blood except in leukocyte count and hemoglobin concentration. The difference in leukocyte counts was thought to depend mainly on the fluctuation of the lymphocyte counts. However, no significant difference was recognized in the proportion of positive cells to each monoclonal antibody. It was concluded that the difference in leukocyte counts found between the arterial and venous bloods of the Wistar-Imamichi rat did not produce any effects on the proportion of the subpopulation in the peripheral lymphocytes, and the lymphocyte subpopulations in both arterial and venous bloods were substantially equivalent to each other.     

End-to-end versus end-to-side microvascular anastomosis patency in experimental venous repairs

In this experimental study, venous end-to-end and end-to-side microvascular anastomoses in similar and diameter-discrepant vessels were compared. In 50 rats, end-to-end microvascular repair of the divided epigastric vein and end-to-side repair of the epigastric vein into the femoral vein showed 5-day patency rates of 75 and 88 percent, respectively. These data are not statistically different. In 20 rats, microvascular repair of end epigastric to end femoral veins (size discrepant) and end epigastric to side femoral veins showed 5-day patency rates of 50 and 85 percent, respectively. These data are statistically different (p less than 0.05). We conclude from these experimental data that end-to-side venous repairs may be useful in lowering the anastomosis thrombosis rate seen when size-discrepant veins are repaired.     

Inflammatory cells induce neointimal growth in a rat arterial autograft model

Subendothelial invasion by leukocytes is a sign of intimal thickening in arteriosclerosis and in the response of a vessel to mechanical damage. Our study was designed to establish whether these cells are implicated in the formation of a neointima in an autologous arterial graft model in the rat and to evaluate the effects of cyclosporin A (CsA). Three study groups were established according to whether the animals were treated with CsA-Cp (Sandimmun), CsA-Et (ethanol vehicle) or received no treatment (control group). Both drug forms were administered (5 mg/kg/day, s.c.) from 4 days prior to surgery until the time of sacrifice. Antibodies specific for lymphocytes (CD4, CD8), monocytes/macrophages-ED1, smooth muscle alpha-actin and the von Willebrand factor (vWF) were used to identify the cells in the grafted arterial wall. In control grafts, the neointima had formed by 2 weeks post-implant. However, the cells comprising this layer generally presented no positivity whatsoever towards the antibodies employed. At 50 days, the new layer was observed to be formed by a vWF-positive endothelium and alpha-actin-positive cells. In all three groups, several polymorphonuclear (PMN) cells adhered to the denuded luminal surface from 7 days onwards. In the treated animals, neutrophils and monocytes were seen to infiltrate intimal and medial layers during the later post-implant stages. Around the third week post-implant, the neointima had reached the grafted segment from the distal portion of the recipient artery, and by 50 days it was similar to that seen in control specimens. Our findings suggest that: a) neutrophils play a role in neointimal thickening in this arterial autograft model; and b) CsA promotes the adhesion and infiltration of neutrophils in the injured arterial wall.     

Pressure drops in a distensible model of end-to-side anastomosis in systemic-to-pulmonary shunts

The modified Blalock-Taussig shunt is a surgical procedure used as a palliation to treat complex congenital heart defects. It consists of an interposing prosthetic tube between the innominate/subclavian artery and the right pulmonary artery. Previous experience indicates that the pressure drop across the shunt is affected by the pulmonary pressure at the distal anastomosis combined with the distensibility of the anastomosis. In this study, a computational fluid-structure interaction approach is presented to investigate the haemodynamic behaviour. Steady-state fluid dynamics and structural analyses were carried out using commercial codes based on the finite element method (FIDAP and ABAQUS) coupled by means of a purposely-developed procedure to transfer boundary conditions. Both prosthetic tube and artery walls were characterised by non-linear material properties. Three different pulmonary pressures (2, 5 and 15 mmHg) and two volume flow rates (0.4 and 0.8 l/min) were investigated. Results indicate that the effects of distensibility at the distal anastomosis on the shunt pressure drop are relevant only when the distal anastomosis on the shunt pressure drop are relevant only when the distal anastomosis is not fully distended, which occurs when the pulmonary pressure is lower than 5 mmHg.     

Preferential use of end-to-side arterial anastomoses in free flap transfers.

In our early clinical experience with free flaps, we used end-to-end arterial anastomoses and in 9 our of 24 we had complete failures--7 of which were due to early arterial thrombosis. Contrarily, in 41 consecutive free flaps with end-to-side anastomoses we have not had a single failure. At the same time that we began using the end-to-side anastomoses, we also began using the latissimus dorsi free flap as our flap of first choice, and we agree that this was probably an additional reason for our improved success rate. The use of end-to-side anastomoses has the following advantages: (1) a high success rate; (2) preservation of all existing vessels in an injured extremity; (3) greater freedom of operative planning; and (4) technical simplicity in terms of access to the vessels. For us, these advantages have made end-to-side anastomosis the technique of choice in the transfer of free flaps.     

The arterial and venous blood supply of the septum pellucidum in the rat.

The arterial and venous systems of the rat's septum pellucidum has been studied by means of the perfusion technique. The arteries may be classified into three groups. The branches of each of the three groups originate from the a. hemispherica and enter the septum from the frontal direction. The veins may be divided into an anterior and a posterior group. The anterior veins flow through the v. subcallosa into the sinus cavernosus, the posterior veins into v. cerebri magna. The arterial and venous blood supply of the individual nuclei of the septum are compar independent.     

Effect of different oxygen saturations on venous versus arterial endothelial cell growth in vitro

Canine carotid arterial and external jugular venous endothelial cells were derived, cutured, passed, and pooled. Three X 10(6) cells were seeded into capped roller-bottles. Three roller-bottes were seeded with venous, and another three with arterial derived endothelial cells for each of three "oxygen saturations" 5, 20 and 50% O2 respectively, and incubated for seven days and then counted. Arterial endothelial cells showed no cell proliferation in high oxygen saturation compared to an 83% increase in cell numbers when oxygen supply had been held to 20%. Venous endothelial cells showed no cell proliferation in medium or high oxygen supply but low oxygen supply (5% O2) gave a 91% increase in cell counts. The small number of roller-bottles investigated does not allow firm conclusions but our results warrant further evaluation.     

Flow and wall shear stress in end-to-side and side-to-side anastomosis of venous coronary artery bypass grafts

Purpose  

Coronary artery bypass graft (CABG) surgery represents the standard treatment of advanced coronary artery disease. Two major types of anastomosis exist to connect the graft to the coronary artery, i.e., by using an end-to-side or a side-to-side anastomosis. There is still controversy because of the differences in the patency rates of the two types of anastomosis. The purpose of this paper is to non-invasively quantify hemodynamic parameters, such as mass flow and wall shear stress (WSS), in end-to-side and side-to-side anastomoses of patients with CABG using computational fluid dynamics (CFD).     

Numerical simulation of wall shear stress conditions and platelet localization in realistic end-to-side arterial anastomoses

Research studies over the last three decades have established that hemodynamic interactions with the vascular surface as well as surgical injury are inciting mechanisms capable of eliciting distal anastomotic intimal hyperplasia (IH) and ultimate bypass graft failure. While abnormal wall shear stress (WSS) conditions have been widely shown to affect vascular biology and arterial wall self-regulation, the near-wall localization of critical blood particles by convection and diffusion may also play a significant role in IH development. It is hypothesized that locations of elevated platelet interactions with reactive or activated vascular surfaces, due to injury or endothelial dysfunction, are highly susceptible to IH initialization and progression. In an effort to assess the potential role of platelet-wall interactions, experimentally validated particle-hemodynamic simulations have been conducted for two commonly implemented end-to-side anastomotic configurations, with and without proximal outflow. Specifically, sites of significant particle interactions with the vascular surface have been identified by a novel near-wall residence time (NWRT) model for platelets, which includes shear stress-based factors for platelet activation as well as endothelial cell expression of thrombogenic and anti-thrombogenic compounds. Results indicate that the composite NWRT model for platelet-wall interactions effectively captures a reported shift in significant IH formation from the arterial floor of a relatively high-angle (30 deg) graft with no proximal outflow to the graft hood of a low-angle graft (10 deg) with 20% proximal outflow. In contrast, other WSS-based hemodynamic parameters did not identify the observed system-dependent shift in IH formation. However, large variations in WSS-vector magnitude and direction, as encapsulated by the WSS-gradient and WSS-angle-gradient parameters, were consistently observed along the IH-prone suture-line region. Of the multiple hemodynamic factors capable of eliciting a hyperplastic response at the cellular level, results of this study indicate the potential significance of platelet-wall interactions coinciding with regions of low WSS in the development of IH.     

Numerical simulations of pulsatile flow in an end-to-side anastomosis model

A potential interaction between the local hemodynamics and the artery wall response has been suggested for vascular graft failure by intimal hyperplasia (IH). Among the various hemodynamic factors, wall shear stress has been implicated as the primary factor responsible for the development of IH. In order to explore the role of hemodynamics in the formation of IH in end-to-side anastomosis, computational fluid dynamics is employed. To validate the numerical simulations, comparisons with existing experimental data are performed for both steady and pulsatile flows. Generally, good agreement is observed with the velocity profiles whereas some discrepancies are found in wall shear stress (WSS) distributions. Using the same end-to-side anastomosis geometry, numerical simulations are extended using a femoral artery waveform to identify the possible role of unsteady hemodynamics. In the current simulations, Carreau-Yasuda model is used to account for the non-Newtonian nature of blood. Computations indicated a disturbed flow field at the artery-graft junction leading to locally elevated shear stresses on the vascular wall. Furthermore, the shear stress distribution followed the same behavior with oscillating magnitude over the entire flow cycle. Thus, distal IH observed in end-to-side artery-graft models may be caused by the fluctuations in WSS's along the wall.     

Ventricular-arterial coupling in a rat model of reduced arterial compliance provoked by hypervitaminosis D and nicotine

The vitamin D(3) and nicotine (VDN) model is one of isolated systolic hypertension (ISH) in which arterial calcification raises arterial stiffness and vascular impedance. The effects of VDN treatment on arterial and cardiac hemodynamics have been investigated; however, a complete analysis of ventricular-arterial interaction is lacking. Wistar rats were treated with VDN (VDN group, n = 9), and a control group (n = 10) was included without the VDN. At week 8, invasive indexes of cardiac function were obtained using a conductance catheter. Simultaneously, aortic pressure and flow were measured to derive vascular impedance and characterize ventricular-vascular interaction. VDN caused significant increases in systolic (138 +/- 6 vs. 116 +/- 13 mmHg, P < 0.01) and pulse (42 +/- 10 vs. 26 +/- 4 mmHg, P < 0.01) pressures with respect to control. Total arterial compliance decreased (0.12 +/- 0.08 vs. 0.21 +/- 0.04 ml/mmHg in control, P < 0.05), and pulse wave velocity increased significantly (8.8 +/- 2.5 vs. 5.1 +/- 2.0 m/s in control, P < 0.05). The arterial elastance and end-systolic elastance rose significantly in the VDN group (P < 0.05). Wave reflection was augmented in the VDN group, as reflected by the increase in the wave reflection coefficient (0.63 +/- 0.06 vs. 0.52 +/- 0.05 in control, P < 0.05) and the amplitude of the reflected pressure wave (13.3 +/- 3.1 vs. 8.4 +/- 1.0 mmHg in control, P < 0.05). We studied ventricular-arterial coupling in a VDN-induced rat model of reduced arterial compliance. The VDN treatment led to development of ISH and provoked alterations in cardiac function, arterial impedance, arterial function, and ventricular-arterial interaction, which in many aspects are similar to effects of an aged and stiffened arterial tree.     

Plexiform-like lesions and increased tissue factor expression in a rat model of severe pulmonary arterial hypertension

Severe pulmonary arterial hypertension (PAH) occurs in idiopathic form and in association with diverse diseases. The pathological hallmarks are distal smooth muscle hypertrophy, obliteration of small pulmonary arteriole lumens, and disorganized cellular proliferation in plexiform lesions. In situ thrombosis is also observed. A detailed understanding of the disease progression has been hampered by the absence of an animal model bearing all the pathological features of human disease. To create a model with these characteristics, we gave young (200-g) rats monocrotaline 1 wk following left pneumonectomy; controls with vehicle treatment or sham operation were also studied. In experimental rats, pulmonary arteries had distal smooth muscle hypertrophy and proliferative perivascular lesions. The lesions had a plexiform appearance, occurred early in disease development, and were composed of cells expressing endothelial antigens. Three-dimensional microangiography revealed severe vascular pruning and disorganized vascular networks. We found that expression of tissue factor (TF), the membrane glycoprotein that initiates coagulation, facilitates angiogenesis, and mediates arterial injury in the systemic circulation, was increased in the pulmonary arterioles and plexiform-like lesions of the rats. TF was also heavily expressed in the vessels and plexiform lesions of humans with pulmonary arterial hypertension. We conclude that plexiform-like lesions can be reproduced in rats, and this model will facilitate experiments to address controversies about the role of these lesions in PAH. Increased TF expression may contribute to the prothrombotic diathesis and vascular cell proliferation typical of human disease.     

Comparison of three different supercharging procedures in a rat skin flap model

A significant clinical problem in reconstructive surgery is partial loss of a pedicled flap. To resolve this problem, various methods of vascular augmentation have been developed; "supercharging" is one of those techniques. A new rat flap model was developed for investigation of the supercharging procedure, and the efficacy of the arterial supercharging method was examined. The purpose of this study was to investigate how an arterial supercharging procedure could generate large flap survival areas with different supercharging positions in rats. On the basis of the vascular anatomical features of rats, a circumferential skin flap from the lower abdomen to the back, measuring 4 x 12 cm, was marked. The flap was divided along the dorsal midline. Forty rats were divided into four experimental groups, as follows: group 1 (control), flaps based only on the deep circumflex iliac artery and vein; group 2, flaps supercharged with the ipsilateral superficial inferior epigastric artery; group 3, flaps supercharged with the contralateral superficial inferior epigastric artery; group 4, flaps supercharged with the contralateral deep circumflex iliac artery. On the fourth postoperative day, the flaps were evaluated with measurements of necrosis and survival areas. Microfil (Flow Tech, Inc., Carver, Mass.) was then injected manually throughout the body, and the vascular changes produced by supercharging were angiographically evaluated. Compared with group 1 (control), the flap survival areas were significantly greater in distally supercharged flaps in groups 3 and 4 (mean flap survival, 91.2 +/- 5.2 percent and 90.5 +/- 10.6 percent, respectively; p < 0.001) and in proximally supercharged flaps in group 2 (45.9 +/- 4.1 percent, p < 0.05). Angiographic assessment of the flaps that survived completely revealed marked dilation of the choke veins among the territories and reorientation of dilated veins along the axes of the flaps. This study suggests that distal arterial supercharging (contralateral superficial inferior epigastric artery or contralateral deep circumflex iliac artery) is more effective than proximal arterial supercharging (ipsilateral superficial inferior epigastric artery) in increasing flap survival. Although the rat skin flap may not be analogous to human flaps, distal arterial supercharging might have useful therapeutic potential in increasing flap survival in clinical practice.