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Renal medullary microcirculation: architecture and exchange.

The architecture of the renal medullary microcirculation is highly specialized. Consistent with their role in countercurrent exchange, the vessels (the vasa recta and the intervening capillaries) have high permeabilities to fluid and small hydrophilic solutes. The urea permeability of the continuous endothelium of the descending vasa recta (DVR) in the outer medulla is greatly enhanced by a urea transporter. Aquaporin channels have also been identified in these vessels. In spite of the absence of lymphatics from the inner medulla, fluid uptake from the interstitial fluid (ISF) through the fenestrated endothelium of the ascending vasa recta (AVR) appears to be driven by differences in hydrostatic and oncotic pressure. Because the AVR have high Lp's [10(-5) cm s-1 (cm H2O)-1] and are mechanically linked to surrounding structures, small increments of ISF pressures above the pressure within the AVR can drive significant volumes of fluid into AVR if ISF volume expands. The lower reflection coefficients to serum albumin of the AVR as compared with the DVR may be important in the clearance of interstitial plasma protein. Recent work on isolated DVR from the outer medulla has revealed that these vessels are capable of vasoconstriction and thus of regulating medullary blood flow.

Animals↗

A new method for blood velocimetry in the microcirculation.

We describe a simple new method for computing two-dimensional velocity vector fields of blood flows in microvascular networks. This method, known as optical flow, requires a time sequence of two or more images obtained by a process such as conventional videomicroscopy. There is no need to distinguish individual cells provided clear variations in the light intensity levels are present in the images. The result of the computation reveals the velocity distribution in the microvascular network and the geometry of the blood vessels. Although various implementations of the optical flow technique exist, we have found that the spatial correlation algorithm of Anandan performs best for microcirculatory flows. This report is an introduction to the application of optical flow to the microcirculation and provides instructions on how to obtain and run the codes.

Blood Circulation Time↗

Intravital microscopy of the peripheral lymph node microcirculation in mice.

OBJECTIVE: The purpose of this study was to develop a model for microscopic in situ observation of the murine peripheral lymph node (LN) microcirculation and to characterize the function of the lymphocyte homing receptor L-selectin (CD62L) and the peripheral node addressin (PNAd). The latter is a high-affinity ligand for L-selectin in LN high endothelial venules (HEV). METHODS: The subiliac (superficial inguinal) LN was microsurgically dissected in anesthetized adult mice. The nodal microvascular architecture and venular hemodynamics were characterized by bright field and epifluorescence video microscopy. L1-2 pre-B cells that were either mock transfected (L1-2Vector) or stably transfected to express human L-selectin (L1-2L-selectin) were labeled fluorescently and injected into a feeding artery. Cell adhesion in LN venules was studied in both the presence and absence of neutralizing monoclonal antibodies (MAb) to PNAd and L-selectin. RESULTS: The preparation allowed a detailed analysis of hemodynamic parameters and leukocyte adhesion in LN microvessels. L1-2Vector cells did not interact with LN microvessels. In contrast, L1-2L-selectin cells rolled efficiently in venules but not in arterioles or capillaries. Rolling was most prominent in subcortical HEV (orders III to V) and was less frequent but consistently detectable in downstream medullary and hilus venules (orders I and II). Rolling interactions were abrogated by MAb DREG-56 to the lectin domain of L-selectin and were markedly reduced by the anti-PNAd MAb MECA-79. CONCLUSIONS: The present study develops a new intravital microscopy model for in vivo visualization of leukocyte interactions with microvessels in murine LN. The preparation permitted an analysis of biophysical and molecular mechanisms of leukocyte adhesion to high endothelial cells. The data support the concept that L-selectin and PNAd are the predominant receptor/ligand pair responsible for lymphocyte rolling in HEV. The model will be useful for high-resolution analysis of intra- and extravascular events in living LN.

Animals↗

The cutaneous microcirculation: ultrastructure and microanatomical organization.

The cutaneous microcirculation is organized as two horizontal plexuses. One is situated 1-1.5 mm below the skin surface, and the other is at the dermal-subcutaneous junction. Ascending arterioles and descending venules are paired as they connect the two plexuses. From the upper layer, arterial capillaries arise to form the dermal papillary loops that represent the nutritive component of the skin circulation. There are sphincter-like smooth muscle cells at the point where the ascending arterioles divide to form the arteriolar component of the upper horizontal plexus. At the dermal subcutaneous junction, there are collecting veins with 2-cusped valves that are oriented to prevent the retrograde flow of blood. Laser Doppler flowmetry (LDF) has demonstrated vasomotion of red cell flux localized to the sites of ascending arterioles. The simultaneous recording by LDF of red cell flux and the concentration of moving red blood cells from individual sites allows one to construct by computer topographic maps of these two valves. The two maps, based on initial studies using correlative skin biopsy specimens, can define 1-mm3 volumes of skin that are predominantly arteriolar in composition, predominantly venular in composition, or essentially devoid of all microvascular elements. The electron and light microscopic features that define the microvascular segments, when coupled with the ability of LDF to define the predominant microvascular segments under the probe, will allow one to study both the mechanisms of normal physiological states and the pathogenetic mechanisms underlying pathological skin disorders in which the microvasculature plays a predominant role.

Arterioles↗

Microcirculation in grafted bone. A chamber technique for vital microscopy of rabbit bone transplants.

A method enabling in vivo observations of the microcirculation in orthotopically transplated bone in the rabbit is presented. The technique permits repeated study of a well-defined bone tissue compartment both before and after grafting to an autogenous or allogenous recipient site, respectively. It yields information about the reorganization of the microvascular system and the resorptive and formative processes of bone remodelling during incorporation of grafted bone. In the autologous experimental series, new capillaries were seen in the bone grafts 4 to 5 days after transplantation. No re-utilization of pre-existing graft vessels was observed.

Animals↗

Wound healing after total elbow replacement in rheumatoid arthritis. Wound complications in 50 cases and laser-Doppler imaging of skin microcirculation.

Wound healing complications, predisposing to deep infection, are common following prosthetic surgery of the elbow. 50 capitellocondylar elbow prostheses were inserted, using a lateral approach, in 42 patients with rheumatoid arthritis. The first 5 elbows were immobilized postoperatively for 5 days and the following 45 elbows for 12 days, because of delayed wound healing in 2 of the first 5 elbows. No wound healing complications were recorded in elbows immobilized for 12 days and elbow motion was not compromised. 5 elbows were investigated with laser-Doppler imaging (LDI) technique, both pre- and postoperatively. Postoperative LDI values were considerably higher than preoperative ones, indicating no impairment of local skin microcirculation. The authors conclude that the lateral approach is safe to use in prosthetic surgery on the elbow. Early mobilization can delay wound healing, but this can be prevented by 2 weeks of postoperative immobilization.

Adult↗

Cigarette smoke and hypoxia induce acute changes in the testicular and cerebral microcirculation.

The acute effects of cigarette smoking and hypoxia on the cerebral and testicular microcirculation were studied in anestethised adult rats. Smoking for 2 min did not influence arterial pO2, pCO2 or pH but it induced an increase in cerebral blood flow by 34% and inhibited vasomotion in the testis for about 1 h. One hour after smoke exposure apnea induced a slight increase in arterial pCO2, a significant decrease in pO2, and an increase in cerebral blood flow (CBF) by 54%. In animals not previously exposed to cigarette smoke apnea increased CBF by 121%, demonstrating that a short-term exposure to tobacco smoke influences the cerebrovascular reactivity for more than one hour. In the testis, apnea resulted in a decreased blood flow by 39% and a complete depression of vasomotion. Breathing 10% O2/90% N2 resulted in moderate hypoxia, a total disappearance of the vasomotion in the testis, a 24% decrease in testicular blood flow, but a 23% increase in CBF.

Animals↗

Microcirculation after cerebral venous occlusions as assessed by laser Doppler scanning.

Research on cerebral venous circulation disturbances (CVCDs) has been limited partly by the paucity of animal models that produce consistent venous infarction. Occlusion of two adjacent cortical veins in rats by means of a photochemical thrombotic technique provides a minimally invasive, clinically relevant, and reproducible model suited to study the pathophysiology of CVCDs. In this study, the effects of venous occlusion on regional cortical blood flow and the brain damage that ensues were evaluated. Cortical vein occlusion was induced by photoactivation of rose bengal via 100-microm fiberoptic illumination. The cerebral venous flow pattern was examined using fluorescence angiography until 90 minutes after venous occlusion, and regional cerebral blood flow (rCBF) was determined at 48 locations by using laser Doppler scanning. Histological damage was assessed 48 hours after vein occlusion. Occlusion of two cortical veins (Group T; seven animals) was compared with single-vein occlusion and its ensuing brain damage (Group S; five animals) and with sham-operated control (five animals). An rCBF reduction occurred 30 minutes after occlusion in Group T and was more extensive than the decrease in Group S after 60 minutes. Observation frequency histograms based on local CBF data obtained in Group T demonstrated that local CBF at some sites decreased to a level below the ischemic threshold within 90 minutes. Six of the seven rats in Group T had a growing venous thrombus with extravasation of fluorescein. The resulting infarction was significantly larger in Group T (9.8 +/- 4.5% of the hemispheric area) than in Group S (only 3 +/- 1.5% of the hemispheric area). In conclusion, microcirculation perturbations occur early after venous occlusion and result in the formation of a venous thrombus accompanied by local ischemia and severe venous infarction. The extent of vein occlusion determines the resulting brain damage. Based on the results of this study, the authors conclude that CVCDs may be attenuated by prevention of venous thrombus progression together with the use of protective measures against the consequences of ischemia.

Animals↗

About the importance of account of some microcirculation parameters dispersion in modelling oxygen transport.

During the recent years various investigators have developed the technique of kryophotometric measurements of oxyhemoglobin saturation with oxygen (psi) in small vessels. In this paper, on the basis of the statistical model of the organ developed by the authors, a theoretical relationship is established between the distributions of measured in various groups of experiments in the capillaries and the distributions of psi in the arterioles and venules. It is shown that for the description of the distribution of psi in a separate multicapillary unit, lying between the arteriole and the venule, it is possible to introduce a special distribution function. In some cases this function may be determined analytically. Statistical distributions of other microcirculation parameters may be included into the statistical model of the organ using the similar procedure described in this paper for the parameters psi a' psi v- arteriolar and venular psi.

Humans↗

Leukocyte kinetics in the microcirculation.

The transport of leukocytes in the microcirculation is specific for the type, size, and the rheological and adhesive properties, the microanatomy of the host organ, and the hemodynamics. The adhesion to the endothelium is determined largely by the degree of activation via chemotactic factors. Leukocyte motion differs from that of red cells or platelets in several respects. When granulocytes enter into capillaries, they are deformed just like red cells. Under normal flow conditions, the time to deform at the entry to capillaries is typically 1,000 times larger than for the red cell, leading to temporary obstruction of the capillaries. After entry, granulocytes move with lower velocity than red cells which causes a cell train formation inside the capillary. At the venular side, the granulocyte is displaced from the center stream toward the endothelium by faster moving red cells. This process leads to systematic attachment of the granulocytes to the endothelium. At a reduced perfusion pressure or in the presence of locally elevated levels of chemotactic factors, the granulocytes may not be able to pass through the capillary network, which leads to microvascular obstruction. Organs with a narrow capillary network may thereby become filters for circulating granulocytes. This event is accompanied in many situations with damage to the host organ.

Cell Adhesion↗

Lymphatic microcirculation in frog lung.

Lymphatic microvessels were microscopically observed on the surface of frog lungs. Magnified images of lymphatic microvessels were recorded on video tapes. The lymphatic microcirculation was studied on a TV monitor at the magnification of 1500 times. 1) valves were observed in lymphatic microvessels, whose diameter was 15 micron, in frog lungs, 2) the valves were incompetent, 3) contained particles repeatedly flowed backwards and forwards in each lymphatic section, 4) after repetition of the movements, particles passed through the outlet valve, 5) particles seldom flowed back through the inlet valve into the preceding section of the lymphatic, 6) the peak flow velocity of particles attained 0.5 mm/sec, and 7) the mean flow velocity was 11 +/- 4 micron on an average and +/- SD, 8) the diameter of a localized portion of the lymphatic microvessels changed periodically.

Animals↗

Dynamic viscous flow in distensible vessels of skeletal muscle microcirculation: application to pressure and flow transients.

Blood flow in the microcirculation of the rat skeletal muscle during transient changes of arterial pressure is analyzed theoretically. Although flow in such small vessels is quasi-steady and has a very low Reynolds number, time-dependent nonuniform flows along the length of the blood vessels can be observed due to vessel distensibility. The governing equations for a single microvessel are derived using previously measured microvessel elasticity, and several solutions to different inflow and outflow pressures and flow conditions are investigated. The results indicate that when such distensible microvessels are subjected to a step increase of arterial pressure, the arterial flow shows a rapid overshoot followed by a progressive decay to steady-state. An arterial step flow induces a different response which takes the form of a monotonically increasing pressure. Pressure and flows are nonuniform along the vessel length during such transients. In-vitro whole organ pressure-flow data are presented in the dilated rat gracilis muscle which qualitatively agree with the theoretical predictions.

Animals↗

Changes of tumor microcirculation after transcatheter arterial chemoembolization: first pass perfusion MR imaging and Chinese ink casting in a rabbit model.

AIM: To observe the change of tumor microcirculation after transcatheter arterial chemoembolization (TACE) with bletilla microspheres by using first pass perfusion MR imaging (FP) and Chinese ink casting. METHODS: VX2 carcinoma cells were surgically implanted into the left and right lobes of liver of 30 New Zealand white rabbits, which were divided into 3 groups at random. Emulsion of lipiodol mixed with mitomycin C, and 5-FU bletilla microspheres were injected into the hepatic artery respectively, and saline was used as control agent. MR imaging was performed with turbo-flash sequence 14 d after tumor implantation and 7 d after interventional therapy. The steepest slopes (SS) of the signal intensity versus time curves were created for quantitative analysis, 7.5% Chinese ink gelatin solution was injected through ascending artery (17 cases) or portal vein (2 cases) for lesion microvessel area (MVA) measurement after the last MRI examination. The correlation between perfusion imaging and MVA was studied blindly. RESULTS: The SS values at the rim of tumor in lipiodol group (mean, 49% per second) and bletilla group (mean, 35% per second) were significantly decreased (P<0.05) as compared with control group (mean, 124% per second), no difference was found between lipiodol and bletilla groups (P>0.05). In lipiodol group, the MVAs (24 974+/-11 836 microm(2)) in the center of the tumor were significantly smaller than those of the control group (35 510+/-15 675 microm(2)) (P<0.05), while the MVAs (80 031+/-22 745 microm(2)) around the tumor were significantly increased because small and dense plexuses appeared around the tumor which correlated to intense reaction of granulation tissue. None of the vessels was seen in the tumor in bletilla group, the peripheral MVAs of the tumor were significantly smaller than those of the control group (P<0.05) and lipiodol group (P<0.05). There was a good correlation between SS and MVAs in control group (r(s), 0.985, P<0.0001) and bletilla group (r(s), 0.743, P<0.05), the correlation was not significant in lipiodol group (r(s), 0.527, P>0.05). CONCLUSION: TACE with bletilla microspheres may enhance its anti-tumor effect by inhibiting the angiogenesis, and FP-MRI provides useful information to assess the TACE effect by depicting tumor vascularization and perfusion.

Animals↗

Microcirculation disturbance affects rats with acute severe pancreatitis following lung injury.

AIM: To study the effects of microcirculation disturbance (MD) on rats with acute severe pancreatitis (ASP). METHODS: We developed ASP rat models, and anatomized separately after 1, 3, 5, 7, and 9 h. We took out blood and did hemorrheologic examination and erythrocyte osmotic fragility test, checked up the water content, capillary permeability, and genetic expression of intercellular adhesion molecule-1 (ICAM-1) in lung tissues, examined the apoptosis degree of blood vessel endothelium while we tested related gene expression of Bax and Bcl-2 in lung tissues. We did the same examination in control group. RESULTS: The viscosity of total blood and plasma, the hematocrit, and the erythrocyte osmotic fragility were all increased. Fibrinogen was decreased. The water content in lung tissues and capillary permeability were increased. Apoptosis degree of blood vessel endothelium was increased too. ICAM-1 genetic expression moved up after 1 h and reached its peak value after 9 h. CONCLUSION: MD plays an important role in ASP following acute lung injury (ALI). The functional damage of blood vessel endothelium, the apoptosis of capillary vessel endothelium, WBC edging-concentration and the increasing of erythrocyte fragility are the main reasons of ALI.

Acute Disease↗

Disturbance of hepatic and intestinal microcirculation in experimental liver cirrhosis.

AIM: To analyze hepatic, mesenteric and mucosal microcirculation and leukocyte-endothelium interaction (LEI) in a rat model with liver cirrhosis. METHODS: Hepatic cirrhosis was induced in Wistar rats by gavage with carbon tetrachloride, and intravital videomicroscopy was performed in liver, mesentery and small intestine mucosa. Special emphasis is given on microcirculatory and morphometric changes during cirrhotic portal hypertension. RESULTS: LEI was influenced significantly in the cirrhotic liver but not in the gut. Blood flow measurement showed significant differences among liver, main mesenteric vessels and the mucosa. The results of our study indicate that liver cirrhosis leads to alterations in hepatic and mesenteric blood flow but not in mucosal blood flow. CONCLUSION: The enhanced inflammatory response in hepatic microvessels may be caused by a decrease of antithrombin III levels and could be responsible for disturbances of organ pathology.

Animals↗

Liver microcirculation after hepatic artery embolization with degradable starch microspheres in vivo.

AIM: To observe the dynamic changes of liver microcirculation in vivo after arterial embolization with degradable starch microspheres (DSM). METHODS: DSM were injected into the proper hepatic artery through a silastic tube inserted retrogradely in gastroduodenal artery (GDA) of SD rats. Fluorescent microscopy was used to evaluate the dynamic changes of blood flow through the terminal portal venules (TPVs), sinusoids and terminal hepatic venules (THVs). The movements of DSM debris were also recorded. Six hours after injection of DSM, percentages of THVs with completely stagnant blood flow were recorded. RESULTS: Two phases of blood flow change were recorded. In phase one: after intra-arterial injection of DSM, slow or stagnant blood flow was immediately recorded in TPVs, sinusoids and THVs. This change was reversible, and blood flow resumed completely. In phase two: after phase one, blood flow in TPVs changed again and three patterns of blood flow were recorded. Six hours after DSM injection, 36.9% +/- 9.2% of THVs were found with completely stagnant blood flow. CONCLUSION: DSM can stop the microcirculatory blood flow in some areas of liver parenchyma. Liver parenchyma supplied by arteries with larger A-P shunt is considered at a higher risk of total microcirculatory blood stagnation after injection of DSM through hepatic artery.

Animals↗

Epidural anaesthesia restores pancreatic microcirculation and decreases the severity of acute pancreatitis.

AIM: To investigate the effect of epidural anaesthesia (EA) on pancreatic microcirculation during acute pancreatitis (AP). METHODS: AP was induced by injection of sodium taurocholate into the pancreatic duct of Sprague-Dawley rats. To realize EA, a catheter was introduced into the epidural space between T7 and T9 and bupivacaine was injected. Microcirculatory flow was measured by laser Doppler flowmetry. Arterial blood gas analyses were performed. At the end of the experiment (<or=5 h), pancreas was removed for histology. The animals were divided into three groups: Group 1 (n=9), AP without EA; Group 2 (n=4), EA without AP; and Group 3 (n=6), AP treated by EA. RESULTS: In Group 1, pancreatic microcirculatory flow prior to AP was 141+/- 39 perfusion units (PU). After AP, microcirculatory flow obviously decreased to 9+/- 6 PU (P<0.05). Metabolic acidosis developed with base excess (BE) of -14+/-3 mmol/L. Histology revealed extensive edema and tissue necrosis. In Group 2, EA did not significantly modify microcirculatory flow. BE remained unchanged and histological analysis showed normal pancreatic tissue. In Group 3, AP initially caused a significant decrease in microcirculatory flow from 155+/-25 to 11+/-7 PU (P<0.05). After initiation of EA, microcirculatory flow obviously increased again to 81+/-31 PU (P<0.05). BE was -6+/-4 mmol/L, which was significantly different compared to Group 1 (P<0.05). Furthermore, histology revealed less extensive edema and necrosis in pancreatic tissue in Group 3 than that in Group 1. CONCLUSION: AP caused dramatic microcirculatory changes within the pancreas, with development of metabolic acidosis and tissue necrosis. EA allowed partial restoration of microcirculatory flow and prevented development of tissue necrosis and systemic complications. Therefore, EA should be considered as therapeutic option to prevent evolution from edematous to necrotic AP.

Acute Disease↗

Impaired microcirculation and tissue oxygenation in human cerebral malaria: a single photon emission computed tomography and near-infrared spectroscopy study.

Serial single photon emission computed tomography (SPECT), near-infrared spectroscopy (NIRS), and transcranial doppler (TCD) sonography examinations were performed to investigate changes of cerebral perfusion and tissue oxygenation in a patient with complicated cerebral malaria that have been acquired in Nigeria. On admission to the Neurologic Intensive Care Unit in Innsbruck, Austria, SPECT and NIRS revealed focal right hemispheric hypoperfusion and decreased oxygen saturation, respectively, correlating exactly to the patient's right hemispheric localizing signs. In contrast, TCD examinations of the basal cerebral vessels revealed normal flow patterns. The patient showed an initial Plasmodium falciparum parasitemia rate of 30% and was cured by intravenous quinine and oral mefloquine therapy. He was discharged without neurologic symptoms. Follow-up SPECT and NIRS examinations revealed regular cerebral perfusion and oxygenation patterns in both cortical hemispheres. In summary, the presented findings provide first evidence that noninvasive SPECT and NIRS may be important diagnostic tools in the evaluation of impaired cerebral microcirculation in patients with P. falciparum malaria.

Adult↗