Microvascular Oxygen Distribution in Awake Hamster Window Chamber Model During Hyperoxia

American Journal of Physiology. Heart and Circulatory Physiology. Oct, 2003  |  Pubmed ID: 12805029

The microvascular effects and hemodynamic events following exposure to normobaric hyperoxia (because of inspiration of 100% O2) were studied in the awake hamster window chamber model and compared with normoxia. Hyperoxia increased arterial blood Po2 to 477.9 +/- 19.9 from 60.0 +/- 1.2 mmHg (P < 0.05). Heart rate and blood pressure were unaltered, whereas cardiac index was reduced from 196 +/- 13 to 144 +/- 31 ml.min-1.kg-1 (P < 0.05) in hyperoxia. Direct measurements in the microcirculation showed there was arteriolar vasoconstriction, reduction of microvascular flow (83% of control, P < 0.05), and functional capillary density (FCD, 74 +/- 16% of control), the latter change being significant (P < 0.05). Calculations of oxygen delivery and oxygen consumption based on the measured changes in microvascular blood flow velocity and diameter and estimates of oxygen saturation corrected for the Bohr effect due to the lowered pH and increased Pco2 showed that oxygen transport in the microvascular network did not change between normal and hyperoxic condition. The congruence of systemic and microvascular hemodynamics events found with hyperoxia suggests that the microvascular findings are common to most tissues in the organism, and that hyperoxia, due to vasoconstriction and the decrease of FCD, causes a maldistribution of perfusion in the microcirculation.

Measurement of the Cardiac Output in Small Animals by Thermodilution

Microvascular Research. Sep, 2003  |  Pubmed ID: 12935765

Cardiac output (CO) measurements based on indicator dilution, microspheres, thermodilution and ultrasonic sensors are not suitable for small animals, because of limited blood volume, high heart rates and small caliber vessels that do not allow probe placement within the heart. We developed a modified thermodilution method to measure CO in awake animals weighing less than 100 g. Under anesthesia, the animal is instrumented with a jugular vein catheter placed proximal to the subclavian vein and a temperature probe in the carotid artery with the thermocouple positioned at the aortic arch. Two days after implantation, room temperature saline is injected (150 microl) into the jugular catheter and the temperature change recorded. This system uses the temperature probe as a digital feedback control: (1) to minimize recirculation error; (2) to adjust baseline temperature, thereby increasing sensitivity to small changes in temperature; and (3) to stabilize animal core temperature. The system was calibrated using a laboratory bench model with anatomically scaled components. CO was measured (n=29) in 16 hamsters (65-115 g), and was linearly related to body weight. Cardiac index (CI=CO/weight) was 197.0 +/- 18.8 (ml/min)/kg. Repeated measurements were made. This technique allows correlating systemic flow changes to be correlated to those measured in the microcirculation of window chamber preparations.

Microcirculatory Changes During Chronic Adaptation to Hypoxia

American Journal of Physiology. Heart and Circulatory Physiology. Nov, 2003  |  Pubmed ID: 14561680

Microcirculatory changes in the window chamber preparation in Syrian golden hamsters, secondary to chronic hypoxia adaptation, are presented herein. Adaptation was attained by keeping animals in a 10% oxygen environment for 1 wk and 5% the following week. The following groups were studied: group 1, adapted to chronic hypoxia and kept in a 5% oxygen environment throughout the experiment; group 2, adapted to chronic hypoxia and kept in a 21% oxygen environment 24 h before and during the experiment; and group 3, control. Adaptation caused venule enlargement and hematocrit increase (68.6 +/- 2.44 in group 1, 70 +/- 2.66 in group 2, and 43.27 +/- 2.30 in group 3; P < 0.05). Whereas heart rate decreased in adapted animals, blood pressure remained constant. Group 1 presented alkalosis, hypocapnia, and hypoxemia. The adapted groups had decreased blood flow velocity in arterioles and veins. We found no difference in microvasculature oxygen tension between groups 2 and 3; however, the number of capillaries with flow was markedly reduced in group 1 but significantly increased in group 2. Our findings suggest that, as an adaptation to hypoxia, erythropoiesis may prove beneficial by increasing blood viscosity and shear stress, leading to vasodilatation, in addition to the increase in oxygen-carrying capacity. Calculations show that oxygen extraction in the tissue of the window chamber model was significantly lowered in adapted animals breathing 5% oxygen, but was unchanged from the control when breathing 21% oxygen, even though blood hemoglobin content was increased from 14.5 +/- 0.07 g/dl at control to 21.04 +/- 1.24 g/dl in the adapted animals (P < 0.05).

Radial Displacement of Red Blood Cells During Hemodilution and the Effect on Arteriolar Oxygen Profile

American Journal of Physiology. Heart and Circulatory Physiology. Mar, 2004  |  Pubmed ID: 14615279

In this study, we assessed the magnitude of the erratic deviations in the radial position of red blood cells (RBCs) in the laminar flow regime of arterioles in a hamster window preparation and the intraluminal Po(2) profile to determine whether this variability affects the intraluminal distribution of oxygen in conditions of normal hematocrit and hemodilution. A gated image intensifier was used to visualize fluorescently labeled RBCs in tracer quantities and obtain multiple measurements of RBC radial and longitudinal positions at time intervals on the order of 5 ms within single arterioles (diameter range 40-95 microm). RBCs in the velocity range of 0.3-14 mm/s exhibit a mean coefficient of variation of velocity of 16.9 +/- 10.5% and a SD of the radial position of 1.98 +/- 0.98 microm. Both quantities were inversely related to hematocrit, and the former was significantly lowered by hemodilution. Our experimental results presented very similar values and shape compared with the intraluminal oxygen profile derived theoretically for normal hematocrit, suggesting that shear-augmented diffusion due to the measured radial displacement of RBCs did not significantly affect oxygen diffusion from blood into the arteriolar vessel wall. Po(2) profiles in the arterioles assumed an increasingly parabolic configuration with increasing levels of hemodilution.

Microlymphatic and Tissue Oxygen Tension in the Rat Mesentery

American Journal of Physiology. Heart and Circulatory Physiology. Mar, 2004  |  Pubmed ID: 14630627

Oxygen phosphorescence quenching was used to measure tissue Po(2) of lymphatic vessels of 43.6 +/- 23.1 microm (mean +/- SD) diameter in tissue locations of the rat mesentery classified according to anatomic location. Lymph and adipose tissue Po(2) were 20.6 +/- 9.1 and 34.1 +/- 7.8 mmHg, respectively, with the difference being statistically significant. Rare microlymphatic vessels in connective tissue not surrounded by microvessels had a Po(2) of 0.8 +/- 0.2 mmHg, whereas the surrounding tissue Po(2) was 3.0 +/- 3.2 mmHg, with both values being significantly lower than those of adipose tissue. Lower of lymph fluid Po(2) relative to the surrounding tissue was also evident in paired measurements of Po(2) in the lymphatic vessels and perilymphatic adipose tissue, which was significantly lower than the Po(2) in paired adipose tissue. The Po(2) of the lymphatic fluid of the mesenteric microlymphatics is consistently lower than that of the surrounding adipose tissue by approximately 11 mmHg; therefore, lymph fluid has the lowest Po(2) of this tissue. The disparity between lymph and tissue Po(2) is attributed to the microlymphatic vessel wall and lymphocyte oxygen consumption.

Microvascular Pressure and Functional Capillary Density in Extreme Hemodilution with Low- and High-viscosity Dextran and a Low-viscosity Hb-based O2 Carrier

American Journal of Physiology. Heart and Circulatory Physiology. Jul, 2004  |  Pubmed ID: 14975932

Blood losses are usually corrected initially by the restitution of volume with plasma expanders and subsequently by the restoration of oxygen-carrying capacity using either a blood transfusion or possibly, in the near future, oxygen-carrying plasma expanders. The present study was carried out to test the hypothesis that high-plasma viscosity hemodilution maintains perfused functional capillary density (FCD) by preserving capillary pressure. Microvascular pressure responses to extreme hemodilution with low- (LV) and high-viscosity (HV) plasma expanders and an exchange transfusion with a polymerized bovine cell-free Hb (PBH) solution were analyzed in the awake hamster window chamber model (n = 26). Systemic hematocrit was reduced from 50% to 11%. PBH produced a greater mean arterial blood pressure than the nonoxygen carriers. FCD was higher after a HV plasma expander (70 +/- 15%) vs. PBH (47 +/- 12%). Microvascular pressure spanning the capillary network was higher after a HV plasma expander (16-19 mmHg) compared with PBH (12-16 mmHg) and a LV plasma expander (11-14 mmHg) but lower than control (22-26 mmHg). FCD was found to be directly proportional to capillary pressure. The use of a HV plasma expander in extreme hemodilution maintained the number of perfused capillaries and tissue perfusion by comparison with a LV plasma expander due to increased mean arterial blood pressure and capillary pressure. The use of PBH increased mean arterial pressure but reduced capillary pressure due to vasoconstriction and did not maintain FCD.

Microvascular PO2 During Extreme Hemodilution with Hemoglobin Site Specifically PEGylated at Cys-93(beta) in Hamster Window Chamber

American Journal of Physiology. Heart and Circulatory Physiology. Oct, 2004  |  Pubmed ID: 15191899

The oxygen transport capacity of nonhypertensive polyethylene glycol (PEG)-conjugated hemoglobin solutions were investigated in the hamster chamber window model. Microvascular measurements were made to determine oxygen delivery in conditions of extreme hemodilution [hematocrit (Hct) 11%]. Two isovolemic hemodilution steps were performed with a 6% Dextran 70 (70-kDa molecular mass) plasma expander until Hct was 35% of control. Isovolemic blood volume exchange was continued using two surface-modified PEGylated hemoglobins (P5K2, P(50) = 8.6, and P10K2, P(50) = 8.3; P(50) is the hemoglobin Po(2) corresponding to its 50% oxygen saturation) until Hct was 11%. P5K2 and P10K2 are PEG-conjugated hemoglobins that maintain most of the hemoglobin allosteric properties and have a cooperativity index of n = 2.2. The effects of these molecular solutions were compared with those obtained in a previous study using MP4, a PEG-modified hemoglobin whose P(50) was 5.4 and cooperativity was 1.2 (Tsai et al., Am J Physiol Heart Circ Physiol 285: H1411-H1419, 2003). Tissue oxygen levels were higher after P5K2 (7.0 +/- 2.5 mmHg) and P10K2 (6.3 +/- 2.3 mmHg) versus MP4 (1.7 +/- 0.5 mmHg) or the nonoxygen carrier Dextran 70 (1.3 +/- 1.2 mmHg). Microvascular oxygen delivery was higher after P5K2 and P10K2 (2.22 and 2.34 ml O(2)/dl blood) compared with MP4 (1.41 ml O(2)/dl blood) or Dextran 70 (0.90 ml O(2)/dl blood); however, all these values were lower than control (7.42 ml O(2)/dl blood). The total hemoglobin in blood was similar in all cases; therefore, the improvement in tissue Po(2) and oxygen delivery appears to be due to the increased cooperativity of the new molecules.

Oxygen Delivery and Consumption in the Microcirculation After Extreme Hemodilution with Perfluorocarbons

American Journal of Physiology. Heart and Circulatory Physiology. Jul, 2004  |  Pubmed ID: 15210452

The oxygen transport capacity of fluorocarbons was investigated in the hamster chamber window model microcirculation to determine the rate at which oxygen is delivered to the tissue in conditions of extreme hemodilution [hematocrit (Hct) 11%]. Hydroxyethlyl starch (HES 200; 200 kDa molecular mass) was used as a plasma expander for two isovolemic hemodilutions performed with 10% HES 200 until a Hct of 65%. A third step reduced the Hct to 75% of baseline and was performed with either HES 200 or a 60% perfluorocarbon (PFC) emulsion. Comparisons of HES 200-only-hemodiluted animals versus 4.2 g/kg PFC emulsion-hemodiluted animals were made at 21% and 100% normobaric oxygen ventilation. It was found that systemic and microvascular oxygen delivery was 25% and 400% higher in the PFC animals compared with HES 200 animals, respectively, showing that PFCs deliver oxygen to the tissue when combined with hyperoxic ventilation in the present experiments, with no evidence of vasoconstriction or impaired microvascular function. Oxygen ventilation (100%) led to a positive base excess for the PFC group (5.5 +/- 2.5 mmol/l) versus a negative balance (-0.8 +/- 1.4 mmol/l) for the HES 200 group, suggesting that microvascular findings corresponded to systemic events.

Oxygen-carrying Blood Substitutes: a Microvascular Perspective

Expert Opinion on Biological Therapy. Jul, 2004  |  Pubmed ID: 15268681

Development of a viable blood substitute began by focusing on recreating the oxygen-carrying capacity of blood, leading to the recognition that haemoglobin (Hb) is presently unequalled for this function. However, as human Hb is the only realistic source of this protein, the production of a blood substitute that solves transfusional blood availability problems and shortages must introduce a multiplying factor between supply of natural blood and blood substitute, while maintaining equivalency of function/efficacy. In other words, a unit of blood should produce several units of equivalent blood substitute. This expansion is now possible because of new understanding of how blood delivers oxygen in the microcirculation and the consequences of reducing oxygen-carrying capacity in haemorrhage. This information is used to provide improved resuscitation capacity and maintenance of tissue metabolism by tailoring the properties of a blood substitute to the task of maintaining microvascular function, rather than oxygen delivery capacity. Resuscitation in an organism that is haemorrhaging requires maintenance perfusion, a process directly linked to the maintenance of adequate levels of shear stress on the endothelium, induced by either increased blood/plasma viscosity or increased blood flow velocity in the microcirculation. This process must also be intimately coupled with the requirement that no portion of the tissue is anoxic. This disparate set of requirements can be satisfied with high viscosity Hb solutions that have high affinity for oxygen, a combination of properties that causes the microcirculation to remain functional, and a requirement that supersedes restoration of oxygen-carrying capacity in the treatment of haemorrhage.

Increased Tissue PO2 and Decreased O2 Delivery and Consumption After 80% Exchange Transfusion with Polymerized Hemoglobin

American Journal of Physiology. Heart and Circulatory Physiology. Dec, 2004  |  Pubmed ID: 15297257

The O2-carrying blood substitute based on polymerized bovine hemoglobin (PBH) was used to determine efficacy in maintaining tissue Po2 after an 80% isovolemic blood exchange leading to a hematocrit of 19% [5.4 g Hb/dl from red blood cells (RBCs) and 6.3 g Hb/dl from PBH]. Effects were studied in terms of O2 delivery, O2 extraction, and tissue Po2 at the microcirculatory level at 1, 12, and 24 h after exchange transfusion in awake hamsters prepared with a window chamber model. At 1 h after exchange, arteriolar and venular diameters were decreased compared with baseline. Arteriolar diameter did not fully recover at 12 h after exchange, but venular diameter returned to normal. At 24 h after exchange, arteriolar and venular diameters were not different from baseline. Combining diameter and flow velocity data allowed us to calculate arteriolar and venular flows. At 1 h after exchange, arteriolar and venular flow was reduced compared with baseline. Arteriolar flow was lower at 12 h after exchange and recovered after 24 h. The number of capillaries with RBC passage [functional capillary density (FCD)] at 1 h after exchange with PBH was significantly lower than baseline. FCD remained decreased at 12 h; at 24 h after exchange transfusion, FCD was fully recovered. Tissue Po2 was maximal at 1 h after exchange and decreased progressively at 12 and 24 h after exchange. O2 release to the tissue was minimal at 1 h and increased at 12 and 24 h after exchange. These results suggest the impairment of tissue O2 metabolism after introduction of PBH into the circulation, which is mitigated as PBH concentration declines.

Resuscitation from Hemorrhagic Shock with MalPEG-albumin: Comparison with MalPEG-hemoglobin

Shock (Augusta, Ga.). Oct, 2004  |  Pubmed ID: 15377891

Our aim was to determine the efficacy of polyethylene glycol-conjugated human albumin (MalPEG-Alb) in restoring circulatory volume after 1 h of hemorrhagic shock. Experiments were performed in the awake condition in the hamster skin fold preparation. Microhemodynamic parameters and tissue Po2 were assessed with intravital microscopy and the use of the phosphorescence quenching technique. One hour after shock induction by withdrawal of 50% of the blood volume, animals were resuscitated with MalPEG-Alb (n = 6). Systemic and microhemodynamic parameters following resuscitation were identical to those obtained with the same protocol using MalPEG-Hb (1). However, parameters related to microvascular oxygen distribution were significantly lower in the MalPEG-Alb group compared with the previous data from the MalPEG-Hb group in that tissue oxygen partial pressure was 5 +/- 2 mmHg (vs. 8 +/- 3 mmHg, P < 0.05), oxygen delivery was reduced to 60 +/- 27% (P < 0.05), and oxygen consumption was reduced to 69 +/- 28% (P < 0.05). Both molecules were matched in composition (4.2 g/dL) and surface chemistry. MalPEG-Alb colloid osmotic pressure was 37 mmHg (vs. 49 mmHg for MalPEG-Hb), and viscosity was 2.7 cP (vs. 2.5 cP for MalPEG-Hb). The present results show that both solutions are efficacious plasma expanders and that the hemoglobin-based solution provides improved oxygen distribution and tissue Po2 in the hamster chamber model.

Hyperosmotic-hyperoncotic Versus Hyperosmotic-hyperviscous: Small Volume Resuscitation in Hemorrhagic Shock

Shock (Augusta, Ga.). Nov, 2004  |  Pubmed ID: 15489635

The aim of this study was to test the effects of using a high-viscosity fluid after small-volume hyperosmotic resuscitation from hemorrhagic shock and to compare this to hyperosmotic followed by hyperoncotic resuscitation. Studies were made in the awake hamster window chamber preparation with the animals subjected to hemorrhage of 50% of blood volume and resuscitated with a small volume of a 7.5% NaCl solution, which was followed within minutes by infusion of 25% of withdrawn volume of either 0.7% or 0.8% alginate solutions (A0.7%, 7.6 cp; and A0.8%, 10.2 cp) or 5% hydroxyethyl starch (HES 5%, 2.1 cp). All modalities of resuscitation returned blood pressure to near baseline values in 5 min, which remained elevated after 90 min with A0.7% and A0.8% but returned to near shock values in 15 min with HES 5%. Microvascular flow and functional capillary density (FCD) followed the same pattern, being significantly higher for the alginate solutions than HES 5% after 90 min. Plasma viscosity 90 min after resuscitation was 2.1 and 2.6 cp for A0.7% and A0.8%, respectively, and 1.1 cP for HES 5%. There was an apparent directly proportional relationship between the concentration of alginate and blood pressure recovery, with blood pressure near normal with A0.8%, and approximately 20 mmHg lower with A0.7%. The recovery of microvascular flow and FCD, although showing a trend toward being more effective with A0.8%, was not significantly different from A0.7% but statistically different and improved relative to HES 5%. The high-viscosity fluids provide a novel small-volume method of resuscitation that maximizes microvascular perfusion for extended periods until surgical control of bleeding is possible. Results show that high-plasma-viscosity resuscitation provides a more consistent and prolonged resuscitation than hyperoncotic treatment. The increase in viscosity presents a gradual recovery in blood pressure and may be used as an alternative for small-volume hypotensive resuscitation, increasing tissue perfusion while potentially limiting hemorrhage in vascular injuries of the major blood vessels.

Microvascular Perfusion Upon Exchange Transfusion with Stored Red Blood Cells in Normovolemic Anemic Conditions

Transfusion. Nov, 2004  |  Pubmed ID: 15504169

Transfusions are intended to augment oxygen-carrying capacity. The ability of fresh and stored red blood cells (RBCs) to maintain microvascular perfusion and oxygen delivery to the tissue has not been directly measured.

Oxygen Distribution and Respiration by the Microcirculation

Antioxidants & Redox Signaling. Dec, 2004  |  Pubmed ID: 15548898

Longitudinal and radial oxygen gradients in the microcirculation due to oxygen release from arterioles show that in some tissues oxygen is primarily supplied by arterioles and secondarily by capillaries. In several tissues, the arteriolar rate of oxygen exit is too large to be explained by diffusion alone, indicating that in these tissues oxygen consumption of the arteriolar wall in vivo is much greater than that shown in in vitro studies of endothelium and vascular smooth muscle, a phenomenon that may be related to the synthesis autocoids by the endothelium in vivo. The functional significance of the high metabolic rate of the arteriolar vessels may be related to the need of providing a metabolic barrier for protecting the parenchymal tissue from high oxygen levels in arterial blood, thus reducing formation of oxygen free radicals in the perivascular tissue, a supposition supported by the finding that the radial oxygen gradient at the microvascular wall and therefore its rate of oxygen consumption are proportional to local blood oxygen partial pressure (pO(2)). Oxygen consumption by the endothelium and/or smooth muscle is also a factor in causing terminal lymphatic pO(2) to have the lowest oxygen level in the tissue, rendering this compartment most vulnerable in hypoxic conditions.

Oxygen Transport by Low and Normal Oxygen Affinity Hemoglobin Vesicles in Extreme Hemodilution

American Journal of Physiology. Heart and Circulatory Physiology. Apr, 2005  |  Pubmed ID: 15563528

The oxygen transport capacity of phospholipid vesicles encapsulating purified Hb (HbV) produced with a Po(2) at which Hb is 50% saturated (P 50 ) of 8 (HbV(8)) and 29 mmHg (HbV(29)) was investigated in the hamster chamber window model by using microvascular measurements to determine oxygen delivery during extreme hemodilution. Two isovolemic hemodilution steps were performed with 5% recombinant albumin (rHSA) until Hct was 35% of baseline. Isovolemic exchange was continued using HbV suspended in rHSA solution to a total [Hb] of 5.7 g/dl in blood. P(50) was modified by coencapsulating pyridoxal 5'-phosphate. Final Hct was 11% for the HbV groups, with a plasma [Hb] of 2.1 +/- 0.1 g/dl after exchange with HbV(8) or HbV(29). A reference group was hemodiluted to Hct 11% with only rHSA. All groups showed stable blood pressure and heart rate. Arterial oxygen tensions were significantly higher than baseline for the HbV groups and the rHSA group and significantly lower for the HbV groups compared with the rHSA group. Blood pressure was significantly higher for the HbV(8) group compared with the HbV(29) group. Arteriolar and venular blood flows were significantly higher than baseline for the HbV groups. Microvascular oxygen delivery and extraction were similar for the HbV groups but lower for the rHSA group (P < 0.05). Venular and tissue Po(2) were statistically higher for the HbV(8) vs. the HbV(29) and rHSA groups (P < 0.05). Improved tissue Po(2) is obtained when red blood cells deliver oxygen in combination with a high- rather than low-affinity oxygen carrier.

Microvascular Oxygen Delivery and Consumption Following Treatment with Verapamil

American Journal of Physiology. Heart and Circulatory Physiology. Apr, 2005  |  Pubmed ID: 15563532

The microvascular distribution of oxygen was studied in the arterioles and venules of the awake hamster window chamber preparation to determine the contribution of vascular smooth muscle relaxation to oxygen consumption of the microvascular wall during verapamil-induced vasodilatation. Verapamil HCl delivered in a 0.1 mg/kg bolus injection followed by a continuous infusion of 0.01 mg.kg(-1).min(-1) caused significant arteriolar dilatation, increased microvascular flow and functional capillary density, and decreased arteriolar vessel wall transmural Po(2) difference. Verapamil caused tissue Po(2) to increase from 25.5 +/- 4.1 mmHg under control condition to 32.0 +/- 3.7 mmHg during verapamil treatment. Total oxygen released by the microcirculation to the tissue remained the same as at baseline. Maintenance of the same level of oxygen release to the tissue, increased tissue Po(2), and decreased wall oxygen concentration gradient are compatible if vasodilatation significantly lowers vessel wall oxygen consumption, which in this model appears to constitute an important oxygen-consuming compartment. These findings show that treatment with verapamil, which increases oxygen supply through vasodilatation, may further improve tissue oxygenation by lowering oxygen consumption of the microcirculation.

Elevated Plasma Viscosity in Extreme Hemodilution Increases Perivascular Nitric Oxide Concentration and Microvascular Perfusion

American Journal of Physiology. Heart and Circulatory Physiology. Apr, 2005  |  Pubmed ID: 15576432

We tested the hypothesis that high-viscosity (HV) plasma in extreme hemodilution causes wall shear stress to be greater than low-viscosity (LV) plasma, leading to enhanced production of nitric oxide (NO). The perivascular concentration of NO was measured in arterioles and venules and the tissue of the hamster chamber window model, subjected to acute extreme hemodilution, with a hematocrit (Hct) of 11% using Dextran 500 (n = 6) or Dextran 70 (n = 5) with final plasma viscosities of 1.99 +/- 0.11 and 1.33 +/- 0.04 cp, respectively. HV plasma significantly increased the periarteriolar, perivenular, and tissue NO concentration by 2.0, 1.9, and 1.4 times the control (n = 7). The NO concentration with LV plasma was not statistically different from control. Arteriolar shear stress was significantly increased in HV plasma relative to LV plasma in arterioles but not in venules. Aortic endothelial NO synthase (eNOS) protein expression was increased with HV plasma but not with LV plasma. There was a weak correlation between perivascular NO concentration and the locally calculated shear stress induced by the procedures, when blood viscosity was corrected according to Hct values previously determined in studies of microvascular Hct distribution. The finding that the periarteriolar and venular NO concentration in HV plasma was the same although arteriolar shear stress was significantly greater than venular shear stress maybe be due to differences in vessel wall metabolism between arterioles and venules and the presence of NO transport through the blood stream in the microcirculation. Results support the concept that in extreme hemodilution HV plasma maintains functional capillary density through a NO-mediated vasodilatation.

Alginate Plasma Expander Maintains Perfusion and Plasma Viscosity During Extreme Hemodilution

American Journal of Physiology. Heart and Circulatory Physiology. Apr, 2005  |  Pubmed ID: 15591096

Extreme hemodilution was performed in the hamster chamber window model using 6% Dextran 70, lowering systemic hematocrit by 60%. Animals were subsequently divided into three groups and hemodiluted to a hematocrit of 11% using 6% Dextran 70, 6% Dextran 500, and a 4% Dextran 70 + 0.7% alginate solution (n = 6 each group). Final plasma viscosities were 1.4 +/- 0.2, 2.2 +/- 0.1, and 2.7 +/- 0.2 cp, respectively, (P < 0.05, high viscosity vs. low viscosity). Blood viscosities were 2.1 +/- 0.2, 2.9 +/- 0.4, and 3.9 +/- 0.3 cp, respectively. The lowest blood and plasma viscosity group had a significantly lower functional capillary density, 37 +/- 16%, whereas the two high-viscosity solutions were 71 +/- 15% and 76 +/- 12% (P < 0.05, high viscosity vs. low viscosity), respectively. Arteriolar and venular flow in the Dextran 500 and alginate groups was higher than baseline (i.e., normal nontreated animals), whereas the low-viscosity group showed a reduction in flow. These microvascular changes were paralleled by changes in base excess, which was negative for the Dextran 70 group and positive for the other groups. However, tissue Po(2) was uniformly low for all groups (average of 1.4 mmHg). Calculation of tissue oxygen consumption in the window chamber based on the microvascular data, flow, and intravascular Po(2) showed that only the alginate + Dextran 70 solution-exchanged animals returned to baseline oxygen consumption, whereas the other groups were lower than baseline (P < 0.05). These results show that hemodilution performed with high-viscosity plasma expanders yields systemic arterial pressures and functional capillary densities that are significantly higher (P < 0.05) than those obtained with 6% Dextran 70, a fluid whose viscosity is similar to that of plasma. A condition for obtaining these results is that the oncotic pressure of the plasma expander be titrated to near normal, so that autotransfusion of fluid from the tissue into the vascular compartment does not reduce the effects of increasing plasma viscosity and increased shear stress on the microvascular wall.

Effects of Extreme Hemodilution with Hemoglobin-based O2 Carriers on Microvascular Pressure

American Journal of Physiology. Heart and Circulatory Physiology. May, 2005  |  Pubmed ID: 15637119

A surface-modified polyethylene glycol-conjugated human hemoglobin (MP4) and alpha alpha-cross-linked human hemoglobin (alpha alpha Hb) were used to restore oxygen carrying capacity in conditions of extreme hemodilution (hematocrit 11%) in the hamster window model preparation. Changes in microvascular function were analyzed in terms of effects on capillary pressure and functional capillary density (FCD). MP4, at 1.0 +/- 0.2 g/dl blood concentration, significantly lowered mean arterial pressure (MAP) below baseline (99.6 +/- 7.6 mmHg) to 82.4 +/- 6.9 mmHg (P < 0.05) and decreased of FCD to 70 +/- 9%. alpha alpha Hb caused a greater recovery in MAP to 94.4 +/- 6.2 mmHg and lowered FCD to 62 +/- 8%. However, differences between alpha alpha Hb and MP4 in FCD were not statistically significant. Capillary pressures were in the ranges of 17-21 mmHg for MP4 and 15-19 mmHg for alpha alpha Hb, with both significantly lower than baseline (P < 0.05). Pressure in 80-microm-diameter arterioles was significantly increased with alpha alpha Hb relative to MP4 (P < 0.05). These results were compared with previous findings on the relation between capillary pressure and FCD; they supported the concept of a relationship between FCD and capillary pressure. Measurement of changes in arteriolar diameter, microvascular blood flow, and FCD show that there was no statistical difference between using alpha alpha Hb and MP4 in extreme hemodilution. Microvascular resistance in arterioles with a diameter range of 70-80 microm showed an increase relative to control with alpha alpha Hb, whereas MP4 caused a decrease.

Oxygen Release from Low and Normal P50 Hb Vesicles in Transiently Occluded Arterioles of the Hamster Window Model

American Journal of Physiology. Heart and Circulatory Physiology. Jun, 2005  |  Pubmed ID: 15681705

A phospholipid vesicle encapsulating Hb [Hb vesicle (HbV)] has been developed as a transfusion alternative. One characteristic of HbV is that the O(2) affinity [Po(2) at which Hb is 50% saturated (P(50))] of Hb can be easily regulated by the amount of the coencapsulated allosteric effector pyridoxal 5'-phosphate. In this study, we prepared two HbVs with different P(50)s (8 and 29 mmHg, termed HbV(8) and HbV(29), respectively) and observed their O(2)-releasing behavior from an occluded arteriole in a hamster skinfold window model. Conscious hamsters received HbV(8) or HbV(29) at a dose rate of 7 ml/kg. In the microscopic view, an arteriole (diameter: 53.0 +/- 6.6 mum) was occluded transcutaneously by a glass pipette on a manipulator, and the reduction of the intra-arteriolar Po(2) 100 mum down from the occlusion was measured by the phosphorescence quenching of preinfused Pd-porphyrin. The baseline arteriolar Po(2) (50-52 mmHg) decreased to about 5 mmHg for all the groups. Occlusion after HbV(8) infusion showed a slightly slower rate of Po(2) reduction compared with that after HbV(29) infusion. The arteriolar O(2) content was calculated at each reducing Po(2) in combination with the O(2) equilibrium curves of HbVs, and it was clarified that HbV(8) showed a significantly slower rate of O(2) release compared with HbV(29) and was a primary source of O(2) (maximum fraction, 0.55) overwhelming red blood cells when the Po(2) was reduced (e.g., <10 mmHg) despite a small dosage of HbV. This result supports the possible utilization of Hb-based O(2) carriers with lower P(50) for oxygenation of ischemic tissues.

Increase Plasma Viscosity Sustains Microcirculation After Resuscitation from Hemorrhagic Shock and Continuous Bleeding

Shock (Augusta, Ga.). Jun, 2005  |  Pubmed ID: 15897809

Resuscitation from hemorrhagic shock (50% of blood volume, BV) followed by continuous bleeding (20% of BV per hour, over the entire observation time, 90 min) was studied in the unanesthetized hamster chamber window model. Blood losses equaled 100% of total BV. A single volume infusion (resuscitation) was performed 60 min after hemorrhage using 25% of the BV with 10% hydroxyethyl starch (HES 200, group HES4), or a mixture of HES 200 with 0.3% or 0.6% (w/v) alginate (groups HES7 and HES10, respectively) leading to solutions with a uniform colloidal oncotic pressure (84-87 mmHg) and viscosities ranging from 3.8 to 9.8 cp. Results showed all solutions to be similar immediately after resuscitation (10-15 min) and diverged after this initial period. The viscosity-enhanced solutions showed improved and longer-lasting effects (90 min) relative to the conventional low viscosity, in terms of sustained arterial blood pressure, microvascular flow, capillary perfusion, and laboratory parameters. All microvascular parameters 90 min after resuscitation with low viscosity fell back to the shock level. Improved recovery obtained with a hyperviscous plasma expander was related to microcirculation shear stress preservation, leading to improve blood flow by lowering peripheral vascular resistance when compared with low viscosity resuscitation. These findings suggest the possibility of using hyperviscous plasma expanders to prolong the period for initial treatment of blood losses and definitive institution therapy.

Early Difference in Tissue PH and Microvascular Hemodynamics in Hemorrhagic Shock Resuscitation Using Polyethylene Glycol-albumin- and Hydroxyethyl Starch-based Plasma Expanders

Shock (Augusta, Ga.). Jul, 2005  |  Pubmed ID: 15988323

The hamster chamber window model was subjected to hemorrhagic shock by the withdrawal of 50% of blood volume (BV). BV was restored 1 h after hemorrhage with a single volume infusion (resuscitation) of 25% BV with polyethylene glycol (PEG)-conjugated bovine serum albumin (Alb) and hydroxyethyl starch (HES). Hemorrhage, shock, and resuscitation were monitored continuously in terms of mean arterial pressure (MAP), microvascular blood flow, capillary perfusion, and tissue pH. Blood samples for laboratory parameters were taken at baseline, shock, and resuscitation. Intravascular and tissue pO2 were assessed after resuscitation, and microvascular oxygen supply and extraction were calculated and corrected for pH effect on hemoglobin saturation. Resuscitation with PEG-Alb restored systemic and microvascular parameters up to the end of the observation period (90 min). HES was identical to PEG-Alb resuscitation during the initial 10 to 15 min, but was not sustained subsequently. The trend of recovery in MAP for HES persisted beyond the time when both function capillary density and tissue pH decreased, thus MAP was not indicative of early microvascular dysfunction. Hemoglobin oxygen saturation estimation showed a significant pH dependence. However, oxygen-dependant parameters corrected for pH varied less than 10% from uncorrected data. Early differences found at the microvascular levels suggest that decisions to amend end-result of resuscitation may be short and on the order of minutes. Furthermore, PEG-Alb appears to provide early and long-term sustained systemic and microvascular recovery when used to restitute perfusion and metabolic conditions after resuscitation from hemorrhagic shock.

Blood Viscosity: a Factor in Tissue Survival?

Critical Care Medicine. Jul, 2005  |  Pubmed ID: 16003089

Extreme Hemodilution with PEG-hemoglobin Vs. PEG-albumin

American Journal of Physiology. Heart and Circulatory Physiology. Dec, 2005  |  Pubmed ID: 16024576

Isovolemic hemodilution to 11% systemic hematocrit was performed in the hamster window chamber model using 6% dextran 70 kDa (Dx 70) and 5% human serum albumin (HSA). Systemic and microvascular effects of these solutions were compared with polyethylene glycol (PEG)-conjugated 5% albumin (MPA) and PEG-conjugated 4.2% Hb (MP4). These studies were performed for the purpose of comparing systemic and microvascular responses of PEG vs. non-PEG plasma expanders and similar oxygen-carrying vs. noncarrying blood replacement fluids. Mean arterial blood pressure was statistically significantly reduced for all groups compared with baseline (P < 0.05), HSA, MPA, and MP4 higher than Dx 70 (P < 0.05). MP4 and MPA had a significantly higher cardiac index than HSA and Dx 70, in addition to a positive base excess. Microvascular blood flow and capillary perfusion were significantly higher for the PEG compounds compared with HSA and Dx 70. Intravascular PO2 for MP4 and MPA was higher in arterioles (P < 0.05) compared with HSA and Dx 70, but there was no difference in either tissue or venular PO2 between groups. Total Hb in the MP4 group was 4.8 +/- 0.4 g/dl, whereas the remaining groups had a range of 3.6-3.8 g/dl. The hemodilution results showed that PEG compounds maintained microvascular conditions with lower concentrations than conventional plasma expanders. Furthermore, microvascular oxygen delivery and extraction in the window chamber tissue were significantly higher for the PEG compounds. MP4 was significantly higher than MPA (P < 0.05) and was not statistically different from baseline, an effect due to the additional oxygen release to the tissue by the Hb MP4.

Effect of Oxygen Consumption by Measuring Method on PO2 Transients Associated with the Passage of Erythrocytes in Capillaries of Rat Mesentery

American Journal of Physiology. Heart and Circulatory Physiology. Oct, 2005  |  Pubmed ID: 16162870

Mathematical models have predicted the existence of Po(2) gradients between erythrocytes in capillaries in the usual case where plasma contributes substantial resistance to oxygen diffusion. According to theoretical predictions, these gradients could be detected as rapid Po(2) fluctuations (erythrocyte-associated transients, EATs) along the capillary. However, verification of a model and correct choice of its parameters can be made only on the basis of direct experimental measurements. We used phosphorescence quenching microscopy to measure Po(2) in 52 capillaries of rat mesentery to obtain plasma Po(2) values 100 times/s at a given point along a capillary. A 532-nm laser generated 10-mus pulses of light, concentrated by a x100 objective, onto a spot 0.9 mum in diameter. The presence of erythrocytes in the excitation region was detected on the basis of phosphorescence amplitude (PA), proportional to the amount of plasma encountered by the laser beam, and on the basis of the intensity of transmitted laser light (LT), detected by a photodiode placed under the capillary. The data revealed correlated waveforms in PA, LT, and Po(2) in capillaries. The magnitude of the Po(2) gradients between erythrocytes and plasma was correlated with average capillary Po(2). EATs in Po(2) were more readily detected in capillaries with relatively low oxygenation. The correlation coefficients between PA and Po(2) for the half of the capillaries (n = 26) below the median Po(2) (mean Po(2) = 17 mmHg; R = -0.72) was higher than that for the other half (mean Po(2) = 39 mmHg; R = -0.38). These results support the theoretical predictions of EATs and plasma Po(2) gradients in capillaries.

Role of Endothelial Nitric Oxide in Microvascular Oxygen Delivery and Consumption

Free Radical Biology & Medicine. Nov, 2005  |  Pubmed ID: 16214038

Nitric oxide (NO) is an important signaling molecule modulating diverse processes such as vasodilation, neurotransmission, long-term potentiation, and immune responses. The endothelium contributes a significant fraction of NO from endothelial NO synthase (eNOS). The objective of this work was to analyze the role of eNOS in the modulation of oxygen supply to the tissues and in adaptation to maintain oxygenation uncompromised. Oxygen delivery and consumption were measured in the microcirculation of homozygous mutant endothelial nitric oxide synthase-deficient (eNOS(-/-)) and wild-type mice. Animals were implanted with a dorsal window chamber, allowing us to assess the intact microvascular system. Hemodynamics and oxygen tension were assessed in the microcirculation of conscious animals. The eNOS(-/-) mice had significantly higher blood pressure and lower heart rate (146 +/- 8 mm Hg, 401 +/- 17 bpm) than wild type (127 +/- 6 mm Hg, 428 +/- 20 bpm). Microvascular hemodynamic parameters were not significantly different between groups. The eNOS(-/-) animals delivered less oxygen to the microcirculation and released more oxygen to the tissue; both differences were statistically significant compared to wild type. The arteriolar vessel wall oxygen gradient, a measure of vascular smooth muscle cells and endothelial cell wall oxygen consumption, was significantly lower for eNOS(-/-) than for wild type, suggesting that the inhibition of eNOS is an antianoxia (oxygen sparing) mechanism. Finally, the findings of the study support the argument that NO availability limits oxygen consumption by the tissue.

Nitric Oxide Regulation of Microvascular Oxygen Exchange During Hypoxia and Hyperoxia

Journal of Applied Physiology (Bethesda, Md. : 1985). Apr, 2006  |  Pubmed ID: 16357070

The objective of this work was to test the hypothesis that the limitation of nitric oxide (NO) availability accentuates microvascular reactivity to oxygen. The awake hamster chamber window model was rendered hypoxic and hyperoxic by ventilation with 10 and 100% oxygen. Systemic and microvascular parameters were determined in the two conditions and compared with normoxia in a group receiving the NO scavenger nitronyl nitroxide and a control group receiving only the vehicle (saline). Mean arterial blood pressure did not change with different gas mixtures during infusion of the vehicle, but it increased significantly in the NO-depleted group. NO scavenging increased the reactivity of microvessels to the changed oxygen supply, causing the arteriolar wall to significantly increase oxygen consumption. Tissue Po2 was correspondingly significantly reduced during NO scavenger infusion. The present findings support the hypothesis that microvascular oxygen consumption is proportional to oxygen-induced vasoconstriction. The effect of oxygen on vascular tone is modulated by NO. As a consequence, NO acts as a regulator of the vessel wall oxygen consumption. The vessel wall consumes oxygen in proportion to the local Po2, and an impairment of NO availability renders the circulation more sensitive to changes in the oxygen supply.

Microvascular Effects Following Treatment with Polyethylene Glycol-albumin in Lipopolysaccharide-induced Endotoxemia

Critical Care Medicine. Jan, 2006  |  Pubmed ID: 16374164

To determine whether resuscitation with polyethylene glycol conjugated bovine serum albumin (2.5% weight/volume) infused at 16 mL/kg/hr (PEG-BSA-16) or at 24 mL/kg/hr (PEG-BSA-24) for 1 hr improves microcirculatory conditions in endotoxemia compared with dextran 70 (6% weight/volume) infused at 24 mL/kg/hr (Dex).

Blood Viscosity Maintains Microvascular Conditions During Normovolemic Anemia Independent of Blood Oxygen-carrying Capacity

American Journal of Physiology. Heart and Circulatory Physiology. Aug, 2006  |  Pubmed ID: 16517943

Responses to exchange transfusion with red blood cells (RBCs) containing methemoglobin (MetRBC) were studied in an acute isovolemic hemodiluted hamster window chamber model to determine whether oxygen content participates in the regulation of systemic and microvascular conditions during extreme hemodilution. Two isovolemic hemodilution steps were performed with 6% dextran 70 kDa (Dex70) until systemic hematocrit (Hct) was reduced to 18% (Level 2). A third-step hemodilution reduced the functional Hct to 75% of baseline by using either a plasma expander (Dex70) or blood adjusted to 18% Hct with all MetRBCs. In vivo functional capillary density (FCD), microvascular perfusion, and oxygen distribution in microvascular networks were measured by noninvasive methods. Methylene blue was administered intravenously to reduce methemoglobin (rRBC), which increased oxygen content with no change in Hct or viscosity from MetRBC. Final blood viscosities after the entire protocol were 2.1 cP for Dex70 and 2.8 cP for MetRBC (baseline, 4.2 cP). MetRBC had a greater mean arterial pressure (MAP) than did Dex70. FCD was substantially higher for MetRBC [82 (SD 6) of baseline] versus Dex70 [38 (SD 10) of baseline], and reduction of methemoglobin to oxyhemoglobin did not change FCD [84% (SD 5) of baseline]. P(O2) levels measured with palladium-meso-tetra(4-carboxyphenyl)porphyrin phosphorescence were significantly changed for Dex70 and MetRBC compared with Level 2 (Hct 18%). Reduction of methemoglobin to oxyhemoglobin partially restored P(O2) to Level 2. Wall shear rate and wall shear stress decreased in arterioles and venules for Dex70 and did not change for MetRBC or rRBC. Increased MAP and shear stress-mediated factors could be the possible mechanisms that improved perfusion flow and FCD after exchange for MetRBC. Thus the fall in systemic and microvascular conditions during extreme hemodilution with low-viscosity plasma expanders seems to be, in part, from the decrease in blood viscosity independent of the reduction in oxygen content.

Can the Effects of Vasoactivity of Molecular Hemoglobin-based Plasma Expanders Be Ignored?

Critical Care Medicine. May, 2006  |  Pubmed ID: 16633264

Plasma Viscosity Regulates Systemic and Microvascular Perfusion During Acute Extreme Anemic Conditions

American Journal of Physiology. Heart and Circulatory Physiology. Nov, 2006  |  Pubmed ID: 16731641

The hamster window chamber model was used to study systemic and microvascular hemodynamic responses to extreme hemodilution with low- and high-viscosity plasma expanders (LVPE and HVPE, respectively) to determine whether plasma viscosity is a factor in homeostasis during extreme anemic conditions. Moderated hemodilution was induced by two isovolemic steps performed with 6% 70-kDa dextran until systemic hematocrit (Hct) was reduced to 18% (level 2). In a third isovolemic step, hemodilution with LVPE (6% 70-kDa dextran, 2.8 cP) or HVPE (6% 500-kDa dextran, 5.9 cP) reduced Hct to 11%. Systemic parameters, cardiac output (CO), organ flow distribution, microhemodynamics, and functional capillary density, were measured after each exchange dilution. Fluorescent-labeled microspheres were used to measure organ (brain, heart, kidney, liver, lung, and spleen) and window chamber blood flow. Final blood and plasma viscosities after the entire protocol were 2.1 and 1.4 cP, respectively, for LVPE and 2.8 and 2.2 cP, respectively, for HVPE (baseline = 4.2 and 1.2 cP, respectively). HVPE significantly elevated mean arterial pressure and CO compared with LVPE but did not increase vascular resistance. Functional capillary density was significantly higher for HVPE [87% (SD 7) of baseline] than for LVPE [42% (SD 11) of baseline]. Increases in mean arterial blood pressure, CO, and shear stress-mediated factors could be responsible for maintaining organ and microvascular perfusion after exchange with HVPE compared with LVPE. Microhemodynamic data corresponded to microsphere-measured perfusion data in vital organs.

The Vascular Wall As a Regulator of Tissue Oxygenation

Current Opinion in Nephrology and Hypertension. Jan, 2006  |  Pubmed ID: 16340669

The development of the phosphorescence quenching oxygen measurement technique has allowed for a simultaneous measurement of intra and perivascular partial pressure oxygen along arteriolar vessels in vivo. Mapping the microvascular distribution and oxygen gradients across the vascular walls using this high-resolution technique reveals the existence of large radial gradients between the vasculature and the tissue, with concomitant longitudinal oxygen loss. Mass balance analysis along vessel segments indicates that the vascular wall has a high rate of oxygen consumption. This review presents the current status of in-vivo studies on the partitioning of oxygen between blood, the vascular wall and the surrounding tissue, thereby positioning an oxygen sink between blood and tissue regulating oxygen release.

Oxygen Release from Arterioles with Normal Flow and No-flow Conditions

Journal of Applied Physiology (Bethesda, Md. : 1985). May, 2006  |  Pubmed ID: 16384838

The rate of oxygen release from arterioles ( approximately 55 microm diameter) was measured in the hamster window chamber model during flow and no-flow conditions. Flow was stopped by microvascular transcutaneous occlusion using a glass pipette held by a manipulator. The reduction of the intra-arteriolar oxygen tension (Po2) was measured by the phosphorescence quenching of preinfused Pd-porphyrin, 100 microm downstream from the occlusion. Oxygen release from arterioles was found to be 53% greater during flow than no-flow conditions (2.6 vs. 1.7 x 10(-5) ml O2.cm(-2).s(-1), P < 0.05). Acute hemodilution with dextran 70 was used to reduce vessel oxygen content, significantly increase wall shear stress (14%, P < 0.05), reduce Hct to 28.4% (SD 1.0) [vs. 48.8% (SD 1.8) at baseline], lower oxygen supply by the arterioles (10%, P < 0.05), and increase oxygen release from the arterioles (39%, P < 0.05). Hemodilution also increased microcirculation oxygen extraction (33% greater than nonhemodilution, P < 0.05) and oxygen consumption by the vessel wall, as shown by an increase in vessel wall oxygen gradient [difference in Po2 between the blood and the tissue side of the arteriolar wall, nonhemodiluted 16.2 Torr (SD 1.0) vs. hemodiluted 18.3 Torr (SD 1.4), P < 0.05]. Oxygen released by the arterioles during flow vs. nonflow was increased significantly after hemodilution (3.6 vs. 1.8 x 10(-5) ml O2.cm(-2).s(-1), P < 0.05). The oxygen cost induced by wall shear stress, suggested by our findings, may be >15% of the total oxygen delivery to tissues by arterioles during flow in this preparation.

Increased Cardiac Output and Microvascular Blood Flow During Mild Hemoconcentration in Hamster Window Model

American Journal of Physiology. Heart and Circulatory Physiology. Jul, 2006  |  Pubmed ID: 16489106

The effect of small hematocrit (Hct) increases on cardiac index (cardiac output/body wt) and oxygen release to the microcirculation was investigated in the awake hamster window chamber model by means of exchange transfusions of homologous packed red blood cells. Increasing Hct between 8 and 13% from baseline increased cardiac index by 5-31% from baseline (P < 0.05) and significantly lowered systemic blood pressure (P < 0.05). The relationship between Hct and cardiac index is described by a second-order polynomial (R2 = 0.84; P < 0.05) showing that Hct increases up to 20% from baseline increase cardiac index, whereas increases over 20% from baseline decrease cardiac index. Combining this data with measurements of blood pressure allowed to determine total peripheral vascular resistance, which was a minimum at 8-13% Hct increase and was described by a second-order polynomial (R2 = 0.83; P < 0.05). Oxygen measurements in arterioles, venules, and the tissue at 8-13% Hct increase were identical to control; thus, as a consequence of increased flow and oxygen-carrying capacity, oxygen delivery and extraction increased, but the change was not statistically significant. Previous results with the same model showed that the observed effects are related to shear stress-mediated release of nitric oxide, an effect that should be also present in the heart microcirculation, leading to increased blood flow, myocardial oxygen consumption, and contractility. We conclude that a minimum viscosity level is necessary for generating the shear stress required for maintaining normal cardiovascular function.

Microvascular Perspective of Oxygen-carrying and -noncarrying Blood Substitutes

Annual Review of Biomedical Engineering. 2006  |  Pubmed ID: 16834558

The development of an alternative to natural blood has evolved from the initial goal of replicating blood properties to the current objective of formulating a fluid that can be used to replace blood while preserving microvascular function and delivering oxygen. The properties of this fluid are counterintuitive and different from blood because it has high viscosity, oxygen affinity, and a low oxygen carrier concentration when compared with blood. The optimal oxygen carrier devised presently is poly-ethylene-conjugated human hemoglobin, a material demonstrated to be vasoinactive and void of the toxicities present in previous hemoglobin formulations. A feature of this material is that it is effective in small quantities, and therefore amplifies the equivalent supply of blood derived from blood donations.

Dissociation of Local Nitric Oxide Concentration and Vasoconstriction in the Presence of Cell-free Hemoglobin Oxygen Carriers

Blood. Nov, 2006  |  Pubmed ID: 16857991

Cell-free hemoglobin's (CFH) high affinity for nitric oxide (NO) could limit CFH's use as an oxygen-carrying blood replacement fluid because it scavenges NO, causing vasoconstriction and hypertension. However, the extent to which perivascular NO levels change following intravascular administration of hemoglobin (Hb) with different molecular dimensions correlates with vasoconstrictive responses in the microcirculation is unknown. The study objective was to determine vasoconstrictive effects following bolus infusions of (1) alphaalpha cross-linked Hb; (2) polymerized bovine Hb; or (3) polyethylene glycol-decorated Hb (PEG-Hb), by measurements of in vivo microvessel diameter, blood flow, perivascular NO concentration, and systemic hemodynamic parameters. All CFHs caused reductions in perivascular NO levels, not correlated to microvascular responses. PEG-Hb (largest molecular volume) maintained blood flow, while the others caused vasoconstriction and reduced perfusion. All solutions increased mean arterial pressure due to vasoconstriction and blood volume expansion, except for PEG-Hb, which increased blood pressure due to blood volume expansion and maintenance of cardiac output. In conclusion, perivascular NO reduction is similar for all Hb solutions because NO binding affinities are similar; however, effects on vascular resistance are related to the type of molecular modification, molecular volume, and oxygen affinity.

Hemorrhagic Shock Resuscitation with Carbon Monoxide Saturated Blood

Resuscitation. Feb, 2007  |  Pubmed ID: 17092627

The response to exchange transfusion with red blood cells (RBCs) saturated with carbon monoxide (CO) in amelioration of microvascular function and providing tissue protection in hemorrhagic shock resuscitation was investigated in the hamster chamber window model. Shock was induced by the withdrawal of 50% of blood volume (BV). Blood volume was restored 1 h after hemorrhage with a single volume infusion (resuscitation) of 25% BV with fresh RBCs (saturated or unsaturated with CO) suspended in human serum albumin (HSA). Hemorrhage, shock and resuscitation were monitored continuously in terms of mean arterial pressure (MAP), microvascular blood flow, capillary perfusion and systemic gas parameters. Eight hours after resuscitation, Annexin V and propidium iodide (PI) were injected into the window chamber to study tissue viability, and labeled cells were observed by using intravital epifluorescence microscopy. TUNEL staining was performed on the tissue to confirm in vivo results. Systemic and microvascular restoration were not different with or without CO up to 90 min after resuscitation. CO concentration decreased over 90 min, increasing oxygen carrying capacity and gradually reoxygenating the tissue. CO saturated blood partially mitigated cell injury at 8 h after resuscitation. The precise cellular mechanisms involved require further elucidation. CO is a novel experimental strategy to improve tissue viability and requires the appropriated preclinical studies to confirm its efficacy.

Microvascular and Systemic Effects Following Top Load Administration of Saturated Carbon Monoxide-saline Solution

Critical Care Medicine. Apr, 2007  |  Pubmed ID: 17334240

To determine how top loads with different doses of carbon monoxide (CO)-saturated saline solutions (CO-saline) affect microvascular and systemic hemodynamics and to delineate the corresponding biochemical mechanisms.

Microvascular and Capillary Perfusion Following Glycocalyx Degradation

Journal of Applied Physiology (Bethesda, Md. : 1985). Jun, 2007  |  Pubmed ID: 17347383

Systemic parameters and microvascular and capillary hemodynamics were studied in the hamster window chamber model before and after hyaluronan degradation by intravenous injection of Streptomyces hyaluronidase (100 units, 40-50 U/ml plasma). Glycocalyx permeation was estimated using fluorescent markers of different molecular size (40, 70, and 2,000 kDa), and electrical charge. Systemic parameters (blood pressure, heart rate, blood gases) and microhemodynamics (vascular tone, velocity, and blood flow) remained statistically unchanged after injection of hyaluronidase, compared with inactivated hyaluronidase. Conversely, capillary hemodynamics were drastically affected. Functional capillary density, the capillaries perfused with red blood cells (RBCs), decreased by 35%, capillary Hct of the remaining functional capillaries increased from 16 to 27%, and penetration of 70-kDa fluorescent marker increased. Furthermore, plasma-only perfused capillaries statistically increased 30 min after hyaluronidase. The decrease in functional capillary density accounted for an increased RBC flux in the remainder of the capillaries, since the same number of RBCs had to traverse a reduced number of capillaries. Flux balances showed a reduction from baseline of 11% for the RBC flux and 20% for the plasma flux after treatment. These discrepancies are within the margin of error of the techniques used and could be explained by accounting for RBC over-velocity compared with plasma. These findings suggest that the decrease in the glycocalyx leads to capillary perfusion impairments.

Excessive Erythrocytosis Does Not Elevate Capillary Oxygen Delivery in Subcutaneous Mouse Tissue

Microcirculation (New York, N.Y. : 1994). Feb, 2007  |  Pubmed ID: 17365666

Acclimatization to reduced environmental oxygen includes erythropoietin-regulated increase in erythrocytes enhancing the blood's oxygen content. However, increased hematocrit levels result in elevated blood viscosity that might impair microcirculation and tissue oxygenation. To assess this oxygen supply to the skin, the authors used erythropoietin overexpressing transgenic mice (tg6) that develop excessive erythrocytosis in an oxygen-independent manner. These animals have been previously reported to elevate their blood viscosity 4-fold.

Is Resuscitation from Hemorrhagic Shock Limited by Blood Oxygen-carrying Capacity or Blood Viscosity?

Shock (Augusta, Ga.). Apr, 2007  |  Pubmed ID: 17414420

Systemic and microvascular hemodynamic responses to volume restoration from hemorrhagic shock were studied in the hamster window chamber model to determine the significance of blood rheological and oxygen transport properties. Moderated hemorrhage was induced by means of arterial controlled bleeding of 50% of the blood volume. The hypovolemic shock state was maintained for 1 h before resuscitation. The animals were resuscitated by infusion of 25% of blood volume using either fresh plasma or blood and were studied for 90 min. Transfusion was performed with either oxygen-carrying fresh red blood cells (RBCs) or non-oxygen-carrying RBCs whose hemoglobin was converted to methemoglobin (MetHb). Systemic parameters, including cardiac output, vital organ blood flow distribution, microvascular hemodynamics, and capillary perfusion (functional capillary density [FCD]), were measured during the resuscitation period. Fluorescent-labeled microspheres were used to measure organ blood flow (brain, heart, kidney, liver, lung, spleen, and window chamber). The blood viscosities at the end of the 90-min period were 2.4 cP after resuscitation with plasma, and 2.9 to 3.0 cP after blood transfusion (baseline, 4.2 cP). Resuscitation with RBCs with or without oxygen-carrying capacity had greater mean arterial pressure than did the plasma resuscitation group. The FCD was substantially higher for RBC transfusions (0.56% +/- 7% of baseline) compared with plasma (46% +/- 7% of baseline), and the presence of MetHb in the fresh RBC did not change the FCD or the microvascular hemodynamics. Oxygen delivery and extraction levels were significantly lower for resuscitation with plasma and MetHb-loaded RBCs compared with oxygen-carrying RBCs. The curtailed recovery of systemic and microvascular conditions after volume restitution with plasma seems to be due to the decrease in blood viscosity. Conversely, the restoration of blood rheological properties improves resuscitation independently of the restitution of oxygen-carrying capacity.

Effects of Erythrocyte Flexibility on Microvascular Perfusion and Oxygenation During Acute Anemia

American Journal of Physiology. Heart and Circulatory Physiology. Aug, 2007  |  Pubmed ID: 17449555

Responses to exchange transfusion using red blood cells (RBCs) with normal and reduced flexibility were studied in the hamster window chamber model during acute moderate isovolemic hemodilution to determine the role of RBC membrane stiffness in microvascular perfusion and tissue oxygenation. Erythrocyte stiffness was increased by 30-min incubation in 0.02% glutaraldehyde solution, and unreacted glutaraldehyde was completely removed. Filtration pressure through 5-microm pore size filters was used to quantify stiffness of the RBCs. Anemic conditions were induced by two isovolemic hemodilution steps using 6% 70-kDa dextran to a hematocrit (Hct) of 18% (moderate hemodilution). The protocol continued with an exchange transfusion to reduce native RBCs to 75% of baseline (11% Hct) with either fresh RBCs (RBC group) or reduced-flexibility RBCs (GRBC group) suspended in 5% albumin at 18% Hct; a plasma expander (6% 70-kDa dextran; Dex70 group) was used as control. Systemic parameters, microvascular perfusion, capillary perfusion [functional capillary density (FCD)], and oxygen levels across the microvascular network were measured by noninvasive methods. RBC deformability for GRBCs was significantly decreased compared with RBCs and moderate hemodilution conditions. The GRBC group had a greater mean arterial blood pressure (MAP) than the RBC and Dex70 groups. FCD was substantially higher for RBC (0.81 +/- 0.07 of baseline) vs. GRBC (0.32 +/- 0.10 of baseline) and Dex70 (0.38 +/- 0.10 of baseline) groups. Microvascular tissue Po(2) was significantly lower for Dex70 and GRBC vs. RBC groups and the moderate hemodilution condition. Results were attributed to decreased oxygen uploading in the lungs and obstruction of tissue capillaries by rigidified RBCs, indicating that the effects impairing RBC flexibility are magnified at the microvascular level, where perfusion and oxygenation may define transfusion outcome.

Deferoxamine Lowers Tissue Damage After 80% Exchange Transfusion with Polymerized Hemoglobin

Antioxidants & Redox Signaling. Mar, 2007  |  Pubmed ID: 17184174

Hemoglobin (Hb) solutions have been proposed as potential substitutes for erythrocytes to maintain oxygen-carrying capacity in situations in which blood is not available. This study investigated systemic and microvascular hemodynamics as well as tissue oxygenation and viability after an 80% exchange transfusion with an oxygen-carrying blood substitute based on polymerized bovine hemoglobin (PBH). Studies were carried in unanesthetized hamsters prepared with a window-chamber model for microcirculation evaluation. Heme iron-mediated injury to the tissue was analyzed by using deferoxamine (an iron chelator), which reduces free iron toxicity. Exchange transfusion led to a significant decrease in hematocrit (Hct) and an increase in plasma Hb, in addition to a significant decrease of arteriolar and venular diameters, flow velocity, and, therefore, microvascular blood flow. Capillary perfusion was severely compromised after exchange, but tissue pO2 increased above baseline, and oxygen extraction was reduced. Apoptotic and necrotic cells increased significantly after the exchange; however, this effect was only partially due to the toxicity of free iron. Iron therapy decreased the microvascular and oxygenation changes but did not fully reverse the adverse effects. Assessment of tissue viability after exchange suggests that chelation treatment in cases of large exchange transfusions with acellular Hb could be potentially beneficial.

Perfluorocarbon in Microcirculation During Ischemia Reperfusion

Journal of the American College of Surgeons. Feb, 2007  |  Pubmed ID: 17254926

The effects of perfluorocarbon (PFC) emulsions administered at a nonhemodiluting dose were studied in the hamster window chamber model to determine the difference in ischemia-reperfusion injury associated with PFC delivery before and after an ischemic episode.

Transfusion Restores Blood Viscosity and Reinstates Microvascular Conditions from Hemorrhagic Shock Independent of Oxygen Carrying Capacity

Resuscitation. Oct, 2007  |  Pubmed ID: 17481796

Systemic and microvascular hemodynamic responses to transfusion of oxygen using functional and non-functional packed fresh red blood cells (RBCs) from hemorrhagic shock were studied in the hamster window chamber model to determine the significance of RBCs on rheological and oxygen transport properties. Moderate hemorrhagic shock was induced by arterial controlled bleeding of 50% of the blood volume, and a hypovolemic state was maintained for 1h. Volume restitution was performed by infusion of the equivalent of 2.5 units of packed cells, and the animals were followed for 90 min. Resuscitation study groups were non-oxygen functional fresh RBCs where the hemoglobin (Hb) was converted to methemoglobin (MetHb) [MetRBC], fully oxygen functional fresh RBCs [OxyRBC] and 10% hydroxyethyl starch [HES] as a volume control solution. Measurement of systemic variables, microvascular hemodynamics and capillary perfusion were performed during the hemorrhage, hypovolemic shock and resuscitation. Final blood viscosities after the entire protocol were 3.8 cP for transfusion of RBCs and 2.9 cP for resuscitation with HES (baseline: 4.2 cP). Volume restitution with RBCs with or without oxygen carrying capacity recovered higher mean arterial pressure (MAP) than HES. Functional capillary density (FCD) was substantially higher for transfusion versus HES, and the presence of MetHb in the fresh RBC did not change FCD or microvascular hemodynamics. Oxygen delivery and extraction were significantly lower for resuscitation with HES and MetRBC compared to OxyRBC. Incomplete re-establishment of perfusion after resuscitation with HES could also be a consequence of the inappropriate restoration of blood rheological properties which unbalance compensatory mechanisms, and appear to be independent of the reduction in oxygen carrying capacity.

New Phosphorescence Quenching Oxygen Measurements Technique Yields Unusual Tissue and Plasma PO2 Distributions

Journal of Applied Physiology (Bethesda, Md. : 1985). May, 2007  |  Pubmed ID: 17483445

Time-dependant Oxygen Partial Pressure in Capillaries and Tissue in the Hamster Window Chamber Model

Antioxidants & Redox Signaling. Jul, 2007  |  Pubmed ID: 17508910

The possibility of a plasma oxygen diffusion barrier implies a significant resistance to oxygen diffusion and the existence of capillary erythrocyte-associated transients of oxygen. This effect was analyzed by measuring intracapillary blood and tissue pO(2) in the hamster chamber window model using a noninvasive intravital microscopy palladium porphyrin phosphorescence decay technique for two set light excitations (high and low). Using high light excitation, intracapillary blood pO(2) was 13.7 +/- 6.1 mm Hg, and increased to 18.0 +/- 4.5 mm Hg for low light excitation. For high light excitation, intracapillary blood pO(2) peaks were in the range of 25-30 mm Hg, and the lowest values were in the range of 5-10 mm Hg. Reducing the excitation provided a more uniform pO(2) ranging 15-25 mm Hg. With temporal reduction in blood capillary pO(2), levels were correlated to the increase in phosphorescent amplitude that corresponded to plasma gaps. Tissue pO(2) measured at low light excitation in the proximity of capillaries was 23.1 +/- 1.8 mm Hg. In conclusion, low intracapillary blood pO(2) measurements at full hematocrit are an artifact, only observed when oxygen consumption by the measurement technique was excessive and/or absorption of the excitation light was increased by the absence of RBCs. These findings suggest that resistance to oxygen diffusion in plasma is a minor factor in tissue oxygenation by capillaries in the hamster model.

Resuscitation from Hemorrhagic Shock with Hydroxyethyl Starch and Coagulation Changes

Shock (Augusta, Ga.). Oct, 2007  |  Pubmed ID: 17558350

Administration of fluids to maintain or restore intravascular volume is a common intervention after hemorrhagic shock, but there is uncertainty whether the choice of fluid significantly influences outcome. Systemic parameters, microvascular perfusion, and functional capillary density were used to characterize resuscitation from hemorrhagic shock with hydroxyethyl starch (HES) of different molecular weights. Studies were made in the hamster window chamber model to determine their effects on blood rheological properties, restoration of perfusion and coagulation changes. Moderate hemorrhagic shock was induced by controlled arterial bleeding of 50% of blood volume, and hypovolemia was maintained for 1 h before resuscitation. Twenty-five percent of blood volume was restituted, and recovery was followed over 60 min. Low-molecular weight (MW) HES (L-HES) 130 kd, degree of substitution (DS) 0.40, and high-MW HES (H-HES) 670 kd, DS 0.75, were used as resuscitation fluids. Microthrombi formation was induced by endothelial laser irradiation. H-HES improved systemic conditions, microcirculatory flow, and metabolic recovery after resuscitation when compared with L-HES. Mean arterial pressure was significantly improved after resuscitation with H-HES compared with L-HES, but lower than baseline and the sham group. Thrombus formation was impaired in both groups after resuscitation compared with sham. There was no difference in microthrombi formation between low- and H-HES for medium and large laser endothelial injuries. Our results indicate that fluid resuscitation with HES may increase the risk of bleeding, but not necessarily caused by the properties (MW and DS) of the colloid. Impairment of thrombus formation seems to be in part related to altered hemodynamics and transport inherent to hemodilution, leading to lowered platelet availability due to hemodilution and increased shear stress at the vessel wall when plasma viscosity is increased. The HES MW does not seem to be a factor in compromising platelet adherence on stimulated endothelium. The longer initial intravascular persistence of H-HES might result in longer-lasting volume effects.

Terminal Lymphatics: the Potential "lethal Corner" in the Distribution of Tissue PO2

Lymphatic Research and Biology. 2007  |  Pubmed ID: 18035934

Terminal lymphatic fluid is the compartment furthest removed from the oxygen supply, and therefore should present the lowest pO(2) in the tissue due to oxygen consumption by the tissue and the lymphatic vessel wall.

Increased Hematocrit and Reduced Blood Pressure Following Control of Glycemia in Diabetes

Clinical Hemorheology and Microcirculation. 2008  |  Pubmed ID: 18094460

In this study we determine the effects of reducing blood glucose on mean arterial blood pressure (MAP) and hematocrit (Hct) in patients with type 2 diabetes who are not responding to conventional treatment in an intensive treatment program 1 year after initiation of treatment. Data on MAP, glucose and Hct was obtained from 21 diabetic type 2 individuals subjected to personalized treatment and compared (paired statistics) to pretreatment conditions. Exclusion criteria were severe retinopathy, diabetic nephropathy, amputation of diabetic foot and increased glucose>50 mg/dl. Treatment was the combined administration of glibenclamide and metformin dosed to obtain a reduction of glucose levels. Exercise and strict adherence to a prescribed diet were prescribed in all cases. One year after initiation of therapy, glucose decreased from 219 +/- 87 to 158+/-51 mg/dl (p<0.002), Hct increased from 41.6 +/- 3.2 to 44.7+/-2.9% (p<0.001) and MAP decreased from 100.6 +/- 11.0 to 94.3+/-7.2 mmHg (p<0.001). There were no statically significant changes in cholesterol and triglyceride concentrations. The patients lost weight (72.5+/-12.6 to 70.3+/-13.0 kg, p<0.001) and lowered blood creatinine concentration from 1.04+/-0.24 to 0.95+/-0.25 mg/dl, p<0.05. The increase in Hct should correspond to an increase in blood viscosity of about 12%, however blood pressure, and presumably vascular resistance, decreased by 6%. It is proposed that these effects are in part related to improved kidney function resulting in increased Hct and blood viscosity which increases vascular wall shear stress and NO bioavailability leading to a vasodilator effect.

Isovolemic Exchange Transfusion with Increasing Concentrations of Low Oxygen Affinity Hemoglobin Solution Limits Oxygen Delivery Due to Vasoconstriction

American Journal of Physiology. Heart and Circulatory Physiology. Nov, 2008  |  Pubmed ID: 18835914

O2-carrying fluids based on hemoglobin (Hb) are in various stages of clinical trials to determine their suitability as O2-carrying plasma expanders. Polymerized Hb solutions are characterized by their vasoactivity, low O2 affinity, oncotic effect, prolonged shelf life, and stability. Physiological responses to facilitated O2 transport after exchange transfusion with polymerized bovine Hb (PBH) were studied in the hamster window chamber model during acute moderate anemia to determine how PBH affects microvascular perfusion and tissue oxygenation. The anemic state [29% hematocrit (Hct)] was induced by hemodilution with a plasma expander (70 kDa dextran). After hemodilution, animals were randomly assigned to different exchange transfusion groups. Study groups were based on the concentration of PBH used, namely: PBH at 13 g Hb/dl [PBH13], PBH diluted to 8 (PBH8) or 4 (PBH4) g Hb/dl in albumin solution at matching colloidal osmotic pressure (COP), and no PBH (only albumin solution) at matching COP (PBH0). Measurement of systemic parameters, microvascular hemodynamics, capillary perfusion, and intravascular and tissue O2 levels was performed at 18% Hct. Restitution of O2-carrying capacity with PBH13 increased arterial pressure and triggered vasoconstriction, low perfusion, and high peripheral resistance. PBH4 and PBH0 exhibited lower arterial pressures compared with PBH13. Exchange transfused animals with PBH8 and PBH4 better maintained perfusion and functional capillary density than PBH13. Blood gas parameters and acid-base balance were recovered proportional to microvascular perfusion. Arterial O2 tensions were improved with PBH4 and PBH8 by preventing O2 precapillary release and increasing O2 reserve. Further studies to establish PBH optimal dosage, efficacy, safety, and its effect on outcome are indicated before Hb-based O2-carrying blood substitutes are implemented in routine practice.

Increased Plasma Viscosity Prolongs Microhemodynamic Conditions During Small Volume Resuscitation from Hemorrhagic Shock

Resuscitation. Jun, 2008  |  Pubmed ID: 18308459

Systemic and microvascular hemodynamic responses to hemorrhagic shock resuscitation with hypertonic saline (HTS, 7.5% NaCl) followed with a small volume of plasma expander were studied in the hamster window chamber model to determine the role of plasma expander viscosity in the acute resuscitation outcome. Moderate hemorrhagic shock was induced by arterial controlled bleeding of 50% of blood volume (BV) and the hypovolemic state was maintained for 1 h. Volume restitution was performed by infusion of HTS, 3.5% of BV followed by 10% of BV plasma expanders. Resuscitation was followed for 90 min. The experimental groups were named based on the plasma expanders infused after the HTS, namely: [Hextend], Hextend (6% Hetastarch 670 kDa in lactated electrolyte solution, 4 cp), [Hextend+V], Hextend with viscosity enhanced by the addition of 0.4% alginate, 8 cp, and [NVR] no volume resuscitation as control group. Measurement of systemic parameters, microvascular hemodynamics and capillary perfusion were performed during hemorrhage, shock and resuscitation. Restitution with Hextend yielded the higher mean arterial pressure (MAP), followed by Hextend+V and NVR. Increasing plasma viscosity did not increase peripheral vascular resistance. Functional capillary density (FCD) was higher for Hextend+V than Hextend and NVR. The level of restoration of acid-base balance correlated with microvascular perfusion and was significantly improved with Hextend+V when compared to Hextend and NVR. These results suggest the importance of restoration of blood rheological properties through enhancing plasma viscosity, influencing the re-establishment of microvascular perfusion during small volume resuscitation from hemorrhagic shock.

Survival Time in Severe Hemorrhagic Shock After Perioperative Hemodilution is Longer with PEG-conjugated Human Serum Albumin Than with HES 130/0.4: a Microvascular Perspective

Critical Care (London, England). 2008  |  Pubmed ID: 18423033

Preoperative hemodilution is an established practice that is applied to reduce surgical blood loss. It has been proposed that polyethylene glycol (PEG) surface decorated proteins such as PEG-conjugated human serum albumin may be used as non-oxygen-carrying plasma expanders. The purpose of this study was to determine whether there is any difference in survival time after severe hemorrhagic shock following extreme hemodilution using a conventional hydroxyethyl starch (HES)-based plasma expander or PEG-albumin.

Microvascular Experimental Evidence on the Relative Significance of Restoring Oxygen Carrying Capacity Vs. Blood Viscosity in Shock Resuscitation

Biochimica Et Biophysica Acta. Oct, 2008  |  Pubmed ID: 18502215

The development of volume replacement fluids for resuscitation in hemorrhagic shock comprises oxygen carrying and non carrying fluids. Non oxygen carrying fluids or plasma expanders are used up to the transfusion trigger, and upon reaching this landmark either blood, and possibly in the near future oxygen carrying blood substitutes, are used. An experimental program in hemorrhagic shock using the hamster chamber window model allowed to compare the relative performance of most fluids proposed for shock resuscitation. This model allows investigating simultaneously the microcirculation and systemic reactions, in the awake condition, in a tissue isolated from the environment. Results from this program show that in general plasma expanders such as Ringer's lactate and dextran 70 kDa do not sufficiently restore blood viscosity upon reaching the transfusion trigger, causing microvascular collapse. This is in part restored by a blood transfusion, independently of the oxygen carrying capacity of red blood cells. These results lead to the proposal that effective blood substitutes must be designed to prevent microvascular collapse, manifested in the decrease of functional capillary density. Achievement of this goal, in combination with the increase of oxygen affinity, significantly postpones the need for a blood transfusion, and lowers the total requirement of restoration of intrinsic oxygen carrying capacity.

Oxygen Transport During Hemodilution with a Perfluorocarbon-based Oxygen Carrier: Effect of Altitude and Hyperoxia

Journal of Applied Physiology (Bethesda, Md. : 1985). Aug, 2008  |  Pubmed ID: 18535127

Oxygen delivery and consumption after hemodilution with a perfluorocarbon-based oxygen carrier (PFCOC) was evaluated at sea level and at 2,600 m above sea level. Fifteen anesthetized rats were subjected to a two-exchange normovolemic hemodilution of 40% of the circulating blood volume each. First exchange was performed with a colloid solution. Second exchange was with 80% PFCOC and 20% colloid. Animals were then ventilated with 100% oxygen. Experiments were performed at barometric pressure of 1.0 atm (sea-level group, n=9) or 0.74 atm (2,600-m group, n=6). Blood gases, hematocrit, fluorocrit, and hemoglobin content were measured at baseline and 15 min after each exchange. After hemodilution, total arterial content was not modified by the PFCOC in either group. In contrast, arteriovenous oxygen difference increased significantly in both groups, as did the oxygen extraction ratio. In the second exchange, although total arterial content was similar between the two groups, the perfluorocarbon and plasma phases contributed significantly more at sea level. Arteriovenous oxygen difference was significantly less at sea level with a higher contribution from the perfluorocarbon and plasma phases. In conclusion, hemodilution with a PFCOC induced changes in oxygen delivery and consumption that differ with altitude. The 2,600-m group exhibited a higher oxygen extraction ratio and arteriovenous oxygen difference, with reduced oxygen delivery and unloading from both the fluorocarbon and plasma phase. Therefore, the efficacy of PFCOCs at 2,600 m above sea level is reduced, and altitude must be taken into account when PFCOCs are used.

Volume Resuscitation from Hemorrhagic Shock with Albumin and HexaPEGylated Human Serum Albumin

Resuscitation. Oct, 2008  |  Pubmed ID: 18621463

The effect of restoring intravascular volume with polyethylene glycol (PEG) conjugated to human serum albumin (PEG-Alb) on systemic parameters and microvascular hemodynamics after hemorrhagic shock resuscitation was studied in the hamster window chamber model. Moderate hemorrhagic shock was induced by controlled arterial bleeding of 50% of blood volume, and hypovolemia was maintained for 1h. Fluid resuscitation was accomplished by infusion of 25% of blood volume and recovery was followed over 90 min. The PEG-Alb (six chains of maleimide phenyl PEG conjugated human serum albumin at 4%) resuscitation group was compared human serum albumin (HSA) at 5% (HSA5) and 10% (HSA10) protein concentrations. Systemic parameters, microvascular perfusion and capillary perfusion (functional capillary density, FCD) were measured by noninvasive methods. Hyperoncotic solutions provided rapid restoration of blood pressure, blood gas parameters and microvascular perfusion. Systemic and microvascular recovery was best and most rapid with PEG-Alb and followed by HSA10 and HSA5. Only recovery with PEG-Alb was sustained beyond 90 min. Hemodynamic functional benefits of PEG-Alb and the potential disadvantages associated with HSA, suggest PEG-Alb as better resuscitation solution.

Balance Between Vasoconstriction and Enhanced Oxygen Delivery

Transfusion. Oct, 2008  |  Pubmed ID: 18631171

Hemoglobin (Hb) solutions are potential alternatives to blood transfusion when native oxygen (O(2))-carrying capacity is lacking. Polymerized bovine Hb (PBH) solutions are characterized by its vasoactivity, low O(2) affinity, oncotic effect, prolonged shelf life, and stability. Responses to facilitated O(2) transport, after exchange transfusion with PBH, were studied in the hamster window chamber model during acute extreme anemia to determine how PBH affects microvascular perfusion and tissue oxygenation.

Blood Pressure Reduction Due to Hemoglobin Glycosylation in Type 2 Diabetic Patients

Vascular Health and Risk Management. 2008  |  Pubmed ID: 19066010

To test the hypothesis that glycosylation of hemoglobin constitutes a risk factor for hypertension.

Lowering of Blood Pressure by Increasing Hematocrit with Non Nitric Oxide Scavenging Red Blood Cells

American Journal of Respiratory Cell and Molecular Biology. Feb, 2008  |  Pubmed ID: 17709601

Isovolemic exchange transfusion of 40% of the blood volume in awake hamsters was used to replace native red blood cells (RBCs) with RBCs whose hemoglobin (Hb) was oxidized to methemoglobin (MetHb), MetRBCs. The exchange maintained constant blood volume and produced different final hematocrits (Hcts), varying from 48 to 62% Hct. Mean arterial pressure (MAP) did not change after exchange transfusion, in which 40% of the native RBCs were replaced with MetRBCs, without increasing Hct. Increasing Hct with MetRBCs lowered MAP by 12 mm Hg when Hct was increased 12% above baseline. Further increases of Hct with MetRBCs progressively returned MAP to baseline, which occurred at 62% Hct, a 30% increase in Hct from baseline. These observations show a parabolic "U" shaped distribution of MAP against the change in Hct. Cardiac index, cardiac output divided by body weight, increased between 2 and 17% above baseline for the range of Hcts tested. Peripheral vascular resistance (VR) was decreased 18% from baseline when Hct was increased 12% from baseline. VR and MAP were above baseline for increases in Hct higher than 30%. However, vascular hindrance, VR normalized by blood viscosity (which reflects the contribution of vascular geometry), was lower than baseline for all the increases in Hct tested with MetRBC, indicating prevalence of vasodilation. These suggest that acute increases in Hct with MetRBCs increase endothelium shear stress and stimulate the production of vasoactive factors (e.g., nitric oxide [NO]). When MetRBCs were compared with functional RBCs, vasodilation was augmented for MetRBCs probably due to the lower NO scavenging of MetHb. Consequently, MetRBCs increased the viscosity related hypotension range compared with functional RBCs as NO shear stress vasodilation mediated responses are greater.

Modulation of Perfusion and Oxygenation by Red Blood Cell Oxygen Affinity During Acute Anemia

American Journal of Respiratory Cell and Molecular Biology. Mar, 2008  |  Pubmed ID: 17884988

Responses to exchange transfusion using red blood cells (RBCs) with modified hemoglobin (Hb) oxygen (O(2)) affinity were studied in the hamster window chamber model during acute anemia to determine its role on microvascular perfusion and tissue oxygenation. Allosteric effectors were introduced in the RBCs by electroporation. Inositol hexaphosphate (IHP) and 5-hydroxymethyl-2-furfural (5HMF) were used to decrease and increase Hb-O(2) affinity. In vitro P50s (partial pressure of O(2) at 50% Hb saturation) were modified to 10, 25, 45, and 50 mm Hg (normal P50 is 32 mm Hg). Allosteric effectors also decreased the Hill coefficient. Anemic condition was induced by isovolemic hemodilution exchanges using 6% dextran 70 kD to 18% hematocrit (Hct). Modified RBCs (at 18% Hct in 5% albumin solution) were infused by exchange transfusion of 35% of blood volume. Systemic parameters, microvascular perfusion, capillary perfusion (functional capillary density, FCD), and microvascular Po(2) levels were measured. RBcs with P50 of 45 mm Hg increased tissue Po(2) and decreased O(2) delivery (Do(2)) and extraction (Vo(2)) and RBCs with P50 of 60 mmHg reduced FCD, microvascular flow, tissue Po(2), Do(2) and Vo(2). Erythrocytes with increased Hb-O(2) affinity maintained hemodynamic conditions, Do(2) and decreased tissue Po(2). This study shows that in an anemic condition, maximal tissue Po(2) does not correspond to maximal Do(2) and Vo(2).

Exogenous Nitric Oxide Induces Protection During Hemorrhagic Shock

Resuscitation. Jun, 2009  |  Pubmed ID: 19362408

This study analyzed the systemic and microvascular hemodynamic changes related to increased nitric oxide (NO) availability during the early phase of hemorrhagic shock. Hemodynamic responses to hemorrhagic shock were studied in the hamster window chamber.

Nonobese, Exercising Children Diagnosed with Dyslipidemia Have Normal C-reactive Protein

Vascular Health and Risk Management. 2009  |  Pubmed ID: 19436676

Nonobese children age 10.4 +/- 1.1 years diagnosed with dyslipidemia (n = 51) were compared to normal children age 10.8 +/- 1.1 years (n = 38). Affected individuals had increased total cholesterol: 223 +/- 23 vs 152 +/- 17 mg/dl, p < 0.001; and decreased high-density lipoprotein-cholesterol: 41.9 +/- 4.1 vs 57.6 +/- 5.7 mg/dl, p < 0.001 and triglycerides: 90.8 +/- 40.5 vs 65.7 +/- 25.0 mg/dl, p < 0.002. Fasting glucose was also significantly elevated (p < 0.02). All other parameters, including blood pressure, were not statistically different between groups. The concentration of C-reactive protein was not statistically different between groups. Analysis of medical records showed that this anomaly may be related to this group (as well as the control group) performing regular, daily exercise. This activity was quantified via a self administered questionnaire, and found to be statistically identical in controls and dyslipidemic individuals. Exercise is associated with the release of antiinflammatory cytokines, therefore our results support the contention that it is a significant factor in promoting health conditions from an early stage in life.

Will Fixing the Vasoactivity Caused by Hemoglobin-based Oxygen Carriers Be Enough?

Critical Care Medicine. Jun, 2009  |  Pubmed ID: 19448469

Hematocrit and Mean Arterial Blood Pressure in Pre- and Postmenopause Women

Vascular Health and Risk Management. 2009  |  Pubmed ID: 19554088

The relationship between mean arterial blood pressure (MAP) and hematocrit (Hct) was studied in pre- and postmenopause women in the city of Durango, Mexico. Premenopause women show a negative trend between parameters that is not statistically significant. MAP and Hct are directly related in postmenopause women (p < 0.01). It is proposed that that this MAP/Hct relationship is in part due to differences in endothelial function where menopause decreases the capacity of the endothelium to respond to increased blood viscosity and shears stress, leading to the increased production of vasodilator mediators to compensate for changes in blood viscosity due to changes in Hct. Comparison with a large group of postmenopause women in the city of Stockholm showed identical trends.

Hexa-thiocarbamoyl Phenyl PEG5K Hb: Vasoactivity and Structure: Influence of Rigidity of the Conjugation Linkage on the PEGylation Induced Plasma Expander-like Solution Properties of PEG-Hb Adducts

The Protein Journal. Jun, 2009  |  Pubmed ID: 19653083

A new hexaPEGylated hemoglobin, (TCP-PEG5K)(6)-Hb (TCP, thiocarbamoyl phenyl) has been developed using PEG-phenyl-isothiocyanate and its vasoactivity and structure has been investigated. Of the six PEG5K chains of (TCP-PEG5K)(6)-Hb, 4 are conjugated to the alpha-amino groups of Hb, and the other 2 chains are distributed on epsilon-amino groups, identified as Lys-40(alpha) (approximately 45%), Lys-56(alpha) (approximately 25%), and Lys-8(beta) (approximately 24%). The studies with hamster infused with a bolus of a 4 gm % solution of (TCP-PEG5K)(6)-Hb equivalent to 10% of their blood volume have established that this new hexaPEGylated Hb is vasoinactive. The viscosity and the colloidal osmotic pressure of (TCP-PEG5K)(6)-Hb at 4% is 1.9 cP and 69.7 mmHg, respectively. The molecular radius of (TCP-PEG5K)(6)-Hb is about 4.6 nm and is significantly smaller than hexaPEGylated Hbs developed using other direct and extension arm facilitated PEGylation platform. The presence of an outside the central cavity intramolecular crosslink, succinimidophenyl-PEG2K between Cys-93(beta, beta') in (TCP-PEG5K)(6)-betabeta-Hb strongly impacts its solution properties. These patterns of influence suggest that the inter-dimeric interactions in (TCP-PEG5K)(6)-Hb is weakened just as with other direct PEGylation platforms, and (SP-PEG5K)(6)-Hb generated by EAF-PEGylation is unique in not inducing this effect. A comparison of the properties of hexaPEGylated Hbs establishes that rigidity of the conjugation linkage between PEG and Hb plays a significant influence on the resultant dictating solution properties and/structure/conformation of PEG-Hb adduct.

Effects of the Molecular Mass of Tense-state Polymerized Bovine Hemoglobin on Blood Pressure and Vasoconstriction

Journal of Applied Physiology (Bethesda, Md. : 1985). Nov, 2009  |  Pubmed ID: 19745190

Despite recent advances in the design of hemoglobin (Hb)-based oxygen carriers (HBOCs), vasoconstriction, presumably caused by nitric oxide (NO) scavenging, vessel wall hyperoxygenation, and/or extravasation, has been identified as the principal road block hampering commercial development of HBOCs. This study was designed to analyze systemic and microvascular responses to the molecular mass and plasma concentration of tense (T)-state polymerized bovine Hb (PolybHb) solutions. Experiments were performed using the hamster window chamber model subjected to successive hypervolemic infusions of T-state PolybHb solutions. PolybHb plasma concentrations were evaluated, namely, 0.5, 1.0 and 1.5 g/dl, respectively. Infusion of PolybHb solutions with molecular mass >500 kDa elicited hypertension and vasoconstriction proportional to the plasma concentration and inversely proportional to the PolybHb cross-link density. However, two high-molecular mass PolybHb solutions, PolybHb(40:1)(high) PolybHb(50:1)(high), did not elicit vasoconstriction at all concentrations studied, whereas PolybHb(50:1)(high) only elicited moderate hypertension at the highest concentration studied. In contrast, infusion of PolybHb solutions with molecular mass <500 kDa elicited significant hypertension and vasoconstriction compared with PolybHb solutions with molecular mass >500 kDa that was proportional to the plasma concentration and inversely proportional to the PolybHb cross-link density. We present promising results for highly cross-linked T-state PolybHb solutions with molecular mass >500 kDa [PolybHb(40:1)(high) PolybHb(50:1)(high)], which supports the concept that HBOC size/molecular mass influences its proximity to the vascular endothelium and molecular diffusivity. The hemodynamics of HBOC within the plasma layer surrounding the abluminal side endothelium regulates NO production and consumption, vessel oxygen flux, and extravasation. Although mechanistically attractive, neither of these hypotheses can be directly tested in vivo and will require further investigation.

Low Dose Nitrite Enhances Perfusion After Fluid Resuscitation from Hemorrhagic Shock

Resuscitation. Dec, 2009  |  Pubmed ID: 19804938

This study determines the systemic and microvascular hemodynamic consequences of administering a low dose sodium nitrite after fluid resuscitation from hemorrhagic shock. Hemodynamic responses to hemorrhagic shock and resuscitation were studied in the hamster window chamber model. Moderated hemorrhage was induced by arterial controlled bleeding of 50% of the blood volume (BV) and the hypovolemic state was maintained for 1h. Volume restitution was performed by infusion of 25% of BV using Hextend (6% Hetastarch 670kDa in lactated electrolyte solution) 10min after fluid resuscitation 100microl of specific concentrations of sodium nitrite were infused. The experimental groups were named based on the nitrite concentration used, namely: 0microM, 10microM and 50microM. Systemic parameters, microvascular hemodynamics and capillary perfusion (functional capillary density, FCD) were followed during entire protocol. Exogenous 10microM nitrite maintained systemic and microhemodynamic conditions post fluid resuscitation from hemorrhagic shock, compared to 50microM or no nitrite. A moderated increase in plasma nitrite during the early phase of resuscitation reversed arteriolar vasoconstriction and increased capillary perfusion and venous return, improving central cardiac function. Nitrite effects on resistance vessels, directly influenced intravascular pressure redistribution, sustained blood flow, and prevented tissue ischemia. In conclusion, increasing nitrite plasma bioavailability after fluid resuscitation from hemorrhagic shock can be a potential therapy to enhance microvascular perfusion and to improve overall outcome.

Nitrate and Nitrite in Biology, Nutrition and Therapeutics

Nature Chemical Biology. Dec, 2009  |  Pubmed ID: 19915529

Inorganic nitrate and nitrite from endogenous or dietary sources are metabolized in vivo to nitric oxide (NO) and other bioactive nitrogen oxides. The nitrate-nitrite-NO pathway is emerging as an important mediator of blood flow regulation, cell signaling, energetics and tissue responses to hypoxia. The latest advances in our understanding of the biochemistry, physiology and therapeutics of nitrate, nitrite and NO were discussed during a recent 2-day meeting at the Nobel Forum, Karolinska Institutet in Stockholm.

Polymerized Bovine Hemoglobin Can Improve Small-volume Resuscitation from Hemorrhagic Shock in Hamsters

Shock (Augusta, Ga.). Mar, 2009  |  Pubmed ID: 18636045

Systemic and microvascular hemodynamic responses to hemorrhagic shock volume resuscitation with hypertonic saline followed by infusion of polymerized bovine hemoglobin (PBH) at different concentrations were studied in the hamster window chamber model to determine the role of plasma oxygen-carrying capacity and vasoactivity during resuscitation. Moderate hemorrhagic shock was induced by arterial controlled bleeding of 50% of blood volume (BV), and a hypovolemic state was maintained for 1 h. Volume was restituted by infusion of hypertonic saline (7.5% NaCl), 3.5% of BV, followed by 10% of BV of PBH at 2 different concentrations. Resuscitation was followed for 90 min and was carried out using 13 gPBH/dL (PBH13), PBH diluted to 4 gPBH/dL in albumin solution at matching colloidal osmotic pressure (PBH4), and an albumin-only solution at matching colloidal osmotic pressure (PBH0). Systemic parameters, microvascular hemodynamics, and functional capillary density were determined during hemorrhage, hypovolemic shock, and resuscitation. The PBH13 caused higher arterial pressure without reverting vasoconstriction and hypoperfusion. The PBH4 and PBH0 had lower MAP and partially reverted vasoconstriction. Only treatment with PBH4 restored perfusion and functional capillary density when compared with PBH13 and PBH0. Blood gas parameters and acid-base balance recovered proportionally to microvascular perfusion. Tissue PO2 was significantly improved in the PBH4 group, showing that limited restoration of oxygen-carrying capacity is beneficial and compensates for the effects of vasoactivity, a characteristic of molecular hemoglobin solutions proposed as blood substitutes.

Microcirculatory Effects of Changing Blood Hemoglobin Oxygen Affinity During Hemorrhagic Shock Resuscitation in an Experimental Model

Shock (Augusta, Ga.). Jun, 2009  |  Pubmed ID: 18948853

Microvascular responses to blood volume restitution using red blood cells (RBCs) with modified hemoglobin (Hb) oxygen affinity were studied in the hamster window chamber model during resuscitation from hemorrhagic shock. Allosteric effectors inositol hexaphosphate and 5-hydroxymethyl-2-furfural were introduced into the RBCs by electroporation to decrease and increase Hb-oxygen affinity. In vitro P50s (partial pressure of oxygen at 50% Hb saturation) were modified to 10 and 50 mmHg (normal P50, 32 mmHg). Awake hamsters were subjected to hemorrhage of 50% of blood volume, followed by a shock period of 1 h, and then resuscitated with 25% blood volume with high or low P50 RBCs (hematocrit, 50%). After resuscitation, base excess was significantly lower than baseline in the high-P50 RBC group (HP50; 0.3 +/- 2 vs. 5.0 +/- 1.7 mM) and MAP was lower than baseline in the low-P50 RBC group (LP50; 93 +/- 6 vs. 109 +/- 6 mM). Arteriolar diameter and flow were significantly lower in the HP50. Functional capillary density in the HP50 was significantly lower than LP50 at 60 and 90 min after resuscitation. There was no significantly difference in arteriolar PO2. Tissue PO2, venular PO2, and oxygen delivery were higher in LP50 than in HP50. There was no significant difference in oxygen extraction. Oxygen extraction ratio (oxygen extraction/oxygen delivery) x 100 was significantly higher in HP50 than in LP50. These results suggest that lowering blood P50 in resuscitation provides improved microvascular function in comparison with higher P50.

Cardiac Mechanoenergetic Cost of Elevated Plasma Viscosity After Moderate Hemodilution

Biorheology. 2010  |  Pubmed ID: 21084746

The purpose of this study was to investigate how plasma viscosity affects cardiac and vascular function during moderate hemodilution. Twelve anesthetized hamsters were hemodiluted by 40% of blood volume with two different viscosity plasma expanders. Experimental groups were based on the plasma expander viscosity, namely: high viscosity plasma expander (HVPE, 6.3 mPa · s) and low viscosity plasma expander (LVPE, 2.2 mPa · s). Left ventricular (LV) function was intracardiacally measured with a high temporal resolution miniaturized conductance catheter and concurrent pressure-volume results were used to calculate different LV indices. Independently of the plasma expander, hemodilution decreased hematocrit to 28% in both groups. LVPE hemodilution reduced whole blood viscosity by 40% without changing plasma viscosity, while HVPE hemodilution reduced whole blood viscosity by 23% and almost doubled plasma viscosity relative to baseline. High viscosity plasma expander hemodilution significantly increased cardiac output, stroke volume and stroke work compared to baseline, whereas LVPE hemodilution did not. Furthermore, an increase in plasma viscosity during moderate hemodilution produced a higher energy transfer per unit volume of ejected blood. Systemic vascular resistance decreased after hemodilution in both groups. Counter-intuitively, HVPE hemodilution showed lower vascular resistance and vascular hindrance than LVPE hemodilution. This result suggests that geometrical changes in the circulatory system are induced by the increase in plasma viscosity. In conclusion, an increase in plasma viscosity after moderate hemodilution directly influenced cardiac and vascular function by maintaining hydraulic power and reducing systemic vascular resistance through vasodilation.

Synthesis and Biophysical Properties of Polymerized Human Serum Albumin

Biotechnology Progress. Nov, 2010  |  Pubmed ID: 21213297

The use of many plasma expanders (PEs) is often limited by undesirable side effects, such as red blood cell (RBC) aggregation (hydroxyethyl starch), nephrotoxicity (dextran), and extravasation (albumin). Despite its natural prevalence in the bloodstream, human serum albumin (HSA) can increase the risk of mortality when administered to patients with increased vascular permeability (i.e., patients suffering from burns, septic shock, and endothelial dysfunction). The harmful extravasation of HSA can be limited by polymerizing HSA to increase its molecular size. In this study, HSA was nonspecifically cross-linked with glutaraldehyde at different cross-link densities by varying the molar ratio of glutaraldehyde to HSA. The results of this study show that the weight-averaged molecular weight (MW), viscosity, and extent of RBC aggregation of polymerized HSA increases with increasing cross-link density, whereas the colloid osmotic pressure (COP) decreases with increasing cross-link density. Interestingly, circular dichroism measurements indicate that the secondary structure of HSA is unaffected by polymerization. Altogether, these results show that glutaraldehyde can effectively cross-link HSA to produce high MW polymers, yielding a novel series of potential PEs that exhibit low COP and high viscosity. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011.

Tissue Oxidative Metabolism After Extreme Hemodilution with PEG-conjugated Hemoglobin

Journal of Applied Physiology (Bethesda, Md. : 1985). Dec, 2010  |  Pubmed ID: 20813980

NADH-localized fluorometry was used as a noninvasive technique to monitor changes in the energy state of intact tissue (muscle and connective tissue), without anesthesia, as a function of blood plasma O(2)-carrying capacity in the hamster window chamber model. Acute moderate isovolemic hemodilution was induced by two isovolemic hemodilution steps: in the first step, 6% 70-kDa dextran (Dex70) was used to induce an acute anemic state (18% Hct); in the second step, exchange transfusion of polyethylene glycol (PEG) maleimide-conjugated Hb (4 g/dl, PEG-Hb) or Dex70 (6 g/dl) was used to reduce erythrocytes to 75% of baseline (11% Hct). PEG-Hb had six copies of PEG (5 kDa) conjugated to each human Hb (0.48 g PEG/g Hb) through extension arm-facilitated chemistry. Systemic parameters, microvascular perfusion, functional capillary density, intravascular and interstitial Po(2), and intracellular NADH fluorescence were monitored. Mean arterial blood pressure after extreme hemodilution was statistically significantly reduced for Dex70 compared with PEG-Hb. The presence of PEG-Hb in the circulation maintained positive acid-base balance. While microvascular blood flows were not different, functional capillary density was significantly higher for PEG-Hb than Dex70. Arteriolar Po(2) was higher in the presence of PEG-Hb than Dex70, but tissue and venular Po(2) were not different. Cellular energy metabolism (intracellular O(2)) in the tissues was improved with PEG-Hb. Moderate hemodilution to 18% Hct (6.4 g Hb/dl) brings tissue O(2) delivery to the verge of inadequacy. Extreme hemodilution to 11% Hct (3.7 g Hb/dl) produces tissue anoxia, and high-O(2)-affinity PEG-Hb (Po(2) at which blood is 50% saturated with O(2) = 4 Torr, 1.1 g Hb/dl) only partially decreases anaerobic metabolism without increasing tissue Po(2).

Effects of Plasma Viscosity Modulation on Cardiac Function During Moderate Hemodilution

Asian Journal of Transfusion Science. Jul, 2010  |  Pubmed ID: 20859509

Previous studies have found that increasing plasma viscosity as whole blood viscosity decrease has beneficial effects in microvascular hemodynamics. As the heart couples with systemic vascular network, changes in plasma and blood viscosity during hemodilution determine vascular pressure drop and flow rate, which influence cardiac function. This study aimed to investigate how changes in plasma viscosity affect on cardiac function during acute isovolemic hemodilution.

Tissue Oxygenation After Exchange Transfusion with Ultrahigh-molecular-weight Tense- and Relaxed-state Polymerized Bovine Hemoglobins

American Journal of Physiology. Heart and Circulatory Physiology. Mar, 2010  |  Pubmed ID: 20061539

Hemoglobin (Hb)-based O(2) carriers (HBOCs) constitute a class of therapeutic agents designed to correct the O(2) deficit under conditions of anemia and traumatic blood loss. The O(2) transport capacity of ultrahigh-molecular-weight bovine Hb polymers (PolybHb), polymerized in the tense (T) state and relaxed (R) state, were investigated in the hamster chamber window model using microvascular measurements to determine O(2) delivery during extreme anemia. The anemic state was induced by hemodilution with a plasma expander (70-kDa dextran). After an initial moderate hemodilution to 18% hematocrit, animals were randomly assigned to exchange transfusion groups based on the type of PolybHb solution used (namely, T-state PolybHb and R-state PolybHb groups). Measurements of systemic parameters, microvascular hemodynamics, capillary perfusion, and intravascular and tissue O(2) levels were performed at 11% hematocrit. Both PolybHbs were infused at 10 g/dl, and their viscosities were higher than nondiluted blood. Restitution of the O(2) carrying capacity with T-state PolybHb exhibited lower arterial pressure and higher functional capillary density compared with R-state PolybHb. Central arterial O(2) tensions increased significantly for R-state PolybHb compared with T-state PolybHb; conversely, microvascular O(2) tensions were higher for T-state PolybHb compared with R-state PolybHb. The increased tissue Po(2) attained with T-state PolybHb results from the larger amount of O(2) released from the PolybHb and maintenance of macrovascular and microvascular hemodynamics compared with R-state PolybHb. These results suggest that the extreme high O(2) affinity of R-state PolybHb prevented O(2) bound to PolybHb from been used by the tissues. The results presented here show that T-state PolybHb, a high-viscosity O(2) carrier, is a quintessential example of an appropriately engineered O(2) carrying solution, which preserves vascular mechanical stimuli (shear stress) lost during anemic conditions and reinstates oxygenation, without the hypertensive or vasoconstriction responses observed in previous generations of HBOCs.

Murine Cerebral Malaria is Associated with a Vasospasm-like Microcirculatory Dysfunction, and Survival Upon Rescue Treatment is Markedly Increased by Nimodipine

The American Journal of Pathology. Mar, 2010  |  Pubmed ID: 20110412

Brain hemodynamics in cerebral malaria (CM) is poorly understood, with apparently conflicting data showing microcirculatory hypoperfusion and normal or even increased blood flow in large arteries. Using intravital microscopy to assess the pial microvasculature through a closed cranial window in the murine model of CM by Plasmodium berghei ANKA, we show that murine CM is associated with marked decreases (mean: 60%) of pial arteriolar blood flow attributable to vasoconstriction and decreased blood velocity. Leukocyte sequestration further decreased perfusion by narrowing luminal diameters in the affected vessels and blocking capillaries. Remarkably, vascular collapse at various degrees was observed in 44% of mice with CM, which also presented more severe vasoconstriction. Coadministration of artemether and nimodipine, a calcium channel blocker used to treat postsubarachnoid hemorrhage vasospasm, to mice presenting CM markedly increased survival compared with artemether plus vehicle only. Administration of nimodipine induced vasodilation and increased pial blood flow. We conclude that vasoconstriction and vascular collapse play a role in murine CM pathogenesis and nimodipine holds potential as adjunctive therapy for CM.

Blood Pressure Directly Correlates with Blood Viscosity in Diabetes Type 1 Children but Not in Normals

Clinical Hemorheology and Microcirculation. 2010  |  Pubmed ID: 20134093

To determine the relationship between mean arterial blood pressure (MAP) and blood viscosity in diabetic type 1 children and healthy controls to investigate whether MAP is independent of blood viscosity in healthy children, and vice versa.

Synthesis, Biophysical Properties and Pharmacokinetics of Ultrahigh Molecular Weight Tense and Relaxed State Polymerized Bovine Hemoglobins

Biomaterials. May, 2010  |  Pubmed ID: 20149433

Hemoglobin-based oxygen carriers (HBOC) are currently being developed as red blood cell (RBC) substitutes for use in transfusion medicine. Despite significant commercial development, late stage clinical results of polymerized hemoglobin (PolyHb) solutions hamper development. We synthesized two types of PolyHbs with ultrahigh molecular weights: tense (T) state PolyHb (M(W)=16.59 MDa and P(50)=41 mmHg) and relaxed (R) state PolyHb (M(W)=26.33 MDa and P(50)=0.66 mmHg). By maintaining Hb in either the T- or R-state during the polymerization reaction, we were able to synthesize ultrahigh molecular weight PolyHbs in distinct quaternary states with no tetrameric Hb, high viscosity, low colloid osmotic pressure and the ability to maintain O(2) dissociation, CO association and NO dioxygenation reactions. The PolyHbs elicited some in vitro RBC aggregation that was less than 6% dextran (500 kDa) but more than 5% human serum albumin. In vitro, T-state PolybHb autoxidized faster than R-state PolybHb as expected from previously reported studies, conversely, when administered to guinea pigs as a 20% exchange transfusion, R-state PolybHb oxidized faster and to a greater extent than T-state PolybHb, suggesting a more complex oxidative processes in vivo. Our findings also demonstrate that T-state PolybHb exhibited a longer circulating half-life, slower clearance and longer systemic exposure time compared to R-state PolybHb.

Low Oxygen-affinity Hemoglobin Solution Increases Oxygenation of Partially Ischemic Tissue During Acute Anemia

Journal of the American College of Surgeons. Mar, 2010  |  Pubmed ID: 20193889

Maintenance of postsurgical tissue oxygenation depends on the ability of the specific tissue to recruit perfusion and oxygen (O(2)) supply. When native O(2)-carrying capacity is lacking, fluids to improve O(2)-carrying capacity based in hemoglobin (Hb) could prevent partially ischemic tissue hypoxia by increasing O(2) release from the remaining red blood cells.

Sustained Release Nitric Oxide from Long-lived Circulating Nanoparticles

Free Radical Biology & Medicine. Aug, 2010  |  Pubmed ID: 20460149

The current limitations of nitric oxide (NO) delivery systems have stimulated an extraordinary interest in the development of compounds that generate NO in a controlled and sustained manner with a heavy emphasis on the treatment of cardiovascular disease states. This work describes the positive physiological response to the infusion of NO-releasing nanoparticles prepared using a new platform based on hydrogel/glass hybrid nanoparticles. When exposed to moisture, these nanoparticles slowly release therapeutic levels of NO, previously generated through thermal reduction of nitrite to NO trapped within the dry particles. The controlled and sustained release of NO observed from these nanoparticles (NO-np) is regulated by its hydration over extended periods of time. In a dose-dependent manner, circulating NO-np both decreased mean arterial blood pressure and increased exhaled concentrations of NO over a period of several hours. Circulating NO-np induced vasodilatation and increased microvascular perfusion during their several hour circulation lifetime. Control nanoparticles (control-np; without nitrite) did not induce changes in arterial pressure, although a decrease in the number of capillaries perfused and an increase in leukocyte rolling and immobilization in the microcirculation were observed. The NO released by the NO-np prevents the inflammatory response observed after infusion of control-np. These data suggest that NO release from NO-np is advantageous relative to other NO-releasing compounds, because it does not depend on chemical decomposition or enzymatic catalysis; it is only determined by the rate of hydration. Based on the observed physiological properties, NO-np has clear potential as a therapeutic agent and as a research tool to increase our understanding of NO signaling mechanisms within the vasculature.

The Variability of Blood Pressure Due to Small Changes of Hematocrit

American Journal of Physiology. Heart and Circulatory Physiology. Sep, 2010  |  Pubmed ID: 20601464

The hematocrit (Hct) of awake hamsters was lowered to 90% of baseline by isovolemic hemodilution using hamster plasma to determine the acute effect of small changes in Hct and blood viscosity on systemic hemodynamics. Mean arterial blood pressure increased, reaching a maximum of about 10% above baseline (8.6 +/- 5.5 mmHg) when Hct decreased 8.4 +/- 1.9% (P < 0.005). Cardiac output increased continuously with hemodilution. These conditions were reached at approximately 60 min after exchange transfusion and remained stationary for 1 h. Peripheral vascular resistance was approximately constant up to a decrease of Hct of about 10% and then fell continuously with lowering Hct. Vascular hindrance or vascular resistance independent of blood viscosity increased by about 20% and remained at this level up to an Hct decrease of 20%, indicating that the vasculature constricted with the lowered Hct. The results for the initial 2-h period are opposite but continuous with previous findings with small increases in Hct. In conclusion, limited acute anemic conditions increase mean arterial blood pressure during the initial period of 2 h, an effect that is quantitatively similar but opposite to the acute increase of Hct during the same period.

Perfusion Vs. Oxygen Delivery in Transfusion with "fresh" and "old" Red Blood Cells: the Experimental Evidence

Transfusion and Apheresis Science : Official Journal of the World Apheresis Association : Official Journal of the European Society for Haemapheresis. Aug, 2010  |  Pubmed ID: 20646963

We review the experimental evidence showing systemic and microvascular effects of blood transfusions instituted to support the organism in extreme hemodilution and hemorrhagic shock, focusing on the use of fresh vs. stored blood as a variable. The question: "What does a blood transfusion remedy?" was analyzed in experimental models addressing systemic and microvascular effects showing that oxygen delivery is not the only function that must be addressed. In extreme hemodilution and hemorrhagic shock blood transfusions simultaneously restore blood viscosity and oxygen carrying capacity, the former being critically needed for re-establishing a functional mechanical environment of the microcirculation, necessary for obtaining adequate capillary blood perfusion. Increased oxygen affinity due to 2,3 DPG depletion is shown to have either no effect or a positive oxygenation effect, when the transfused red blood cells (RBCs) do not cause additional flow impairment due to structural malfunctions including increased rigidity and release of hemoglobin. It is concluded that fresh RBCs are shown to be superior to stored RBCs in transfusion, however increased oxygen affinity may be a positive factor in hemorrhagic shock resuscitation. Although experimental studies seldom reproduce emergency and clinical conditions, nonetheless they serve to explore fundamental physiological mechanisms in the microcirculation that cannot be directly studied in humans.

The Physics of Oxygen Delivery: Facts and Controversies

Antioxidants & Redox Signaling. Mar, 2010  |  Pubmed ID: 19757988

Cardiac Systolic Function Recovery After Hemorrhage Determines Survivability During Shock

The Journal of Trauma. Apr, 2011  |  Pubmed ID: 20805773

Small animal model has not been available to study cardiac pathophysiology during hemorrhagic shock. The main purpose of this study, therefore, was to establish earlier differences in left ventricle functional disturbances during hypovolemia comparable in survival and nonsurvival animals. Ventricular pressure-volume relationships have become well established as the most rigorous and comprehensive venue to assess intact heart function.

Biophysical Properties and Oxygenation Potential of High-molecular-weight Glutaraldehyde-polymerized Human Hemoglobins Maintained in the Tense and Relaxed Quaternary States

Tissue Engineering. Part A. Apr, 2011  |  Pubmed ID: 20979534

Recent clinical evaluation of commercial glutaraldehyde-polymerized hemoglobins (PolyHbs) as transfusion solutions has demonstrated several adverse side effects. Chief among these is the hypertensive effect. Fortunately, previous studies have shown that the hypertensive effect can be attenuated by removing free hemoglobin (Hb) and low-molecular-weight (low-MW) PolyHbs from the PolyHb mixture. In this work, polymerized human Hb (PolyhHb) solutions were synthesized in two distinct quaternary states with high MW and subjected to extensive diafiltration to remove free Hb and low-MW PolyhHb components (<500 kDa). The resultant PolyhHb solutions possessed high MW, distinct quaternary state, distinct reactivities with O(2) and CO, similar NO deoxygenating rate constants, distinct autoxidation rate constants, high viscosity, and low colloid osmotic pressure. To preliminarily assess the ability of PolyhHb solutions to oxygenate surrounding tissues fed by a blood vessel, we evaluated the ability of PolyhHbs to transport O(2) to cultured hepatocytes in a mathematical model of a hollow fiber bioreactor. The structure of individual hollow fibers in the bioreactor is similar to that of a blood vessel and provides an easy way to assess the oxygenation potential of PolyhHbs without the need for expensive and time-consuming animal studies. It was observed that PolyhHbs with low O(2) affinities were more effective in oxygenating cultured hepatocytes inside the bioreactor than high O(2) affinity PolyhHbs. Taken together, our results show that it is possible to synthesize high-MW PolyhHbs with no free Hb and low-MW PolyhHb components that are capable of transporting O(2) to cultured cells/tissues.

Reversal of Hemoglobin-induced Vasoconstriction with Sustained Release of Nitric Oxide

American Journal of Physiology. Heart and Circulatory Physiology. Jan, 2011  |  Pubmed ID: 21057038

Erythrocyte free hemoglobin (Hb) induces vasoconstriction due to nitric oxide (NO) scavenging, limiting the NO available for vascular smooth muscle. The central objective of this study was to restore NO bioavailability using long-lived circulating NO-releasing nanoparticles (NO-np) to reverse the vasoconstriction and hypertension induced by polymerized bovine Hb (PBH) NO scavenging. PBH (13 g/dl) was infused in a volume equal to 10% of the animal blood volume. Intravascular NO supplementation was provided with an infusion of NO-np (10 and 20 mg/kg body wt). This study was performed using the hamster window chamber model to concurrently access systemic and microvascular hemodynamics. Infusion of PBH increased blood pressure and induced vasoconstriction. Treatment with 10 and 20 mg/kg NO-np reduced the blood pressure and vasoconstriction induced by PBH. Moreover, the higher dose of NO-np decreased blood pressure and induced vasodilation compared with baseline, respectively. Treatment with NO-np to decrease PBH-induced vasoconstriction increased methemoglobin levels and plasma nitrite and nitrate. In conclusion, NO-np counteracted both systemic hypertension and decreased the vasoconstrictor effects of PBH infusion, improving systemic and microvascular function. Based on the observed physiological properties, NO-np has clear potential as a therapeutic agent to replenish NO in situations where NO production is impaired, insufficient, or consumed, thereby preventing vascular complications.

Small-volume Resuscitation from Hemorrhagic Shock with Polymerized Human Serum Albumin

The American Journal of Emergency Medicine. Nov, 2011  |  Pubmed ID: 22100469

Human serum albumin (HSA) is used as a plasma expander; however, albumin is readily eliminated from the intravascular space. The objective of this study was to establish the effects of various-sized polymerized HSAs (PolyHSAs) during small-volume resuscitation from hemorrhagic shock on systemic parameters, microvascular hemodynamics, and functional capillary density in the hamster window chamber model. Polymerized HSA size was controlled by varying the cross-link density (ie, molar ratio of glutaraldehyde to HSA). Hemorrhage was induced by controlled arterial bleeding of 50% of the animal's blood volume (BV), and hypovolemic shock was maintained for 1 hour. Resuscitation was implemented in 2 phases, first, by infusion of 3.5% of the BV of hypertonic saline (7.5% NaCl) then followed by infusion of 10% of the BV of each PolyHSA. Resuscitation provided rapid recovery of blood pressure, blood gas parameters, and microvascular perfusion. Polymerized HSA at a glutaraldehyde-to-HSA molar ratio of 60:1 (PolyHSA(60:1)) provided superior recovery of blood pressure, microvascular blood flow, and functional capillary density, and acid-base balance, with sustained volume expansion in relation to the volume infused. The high molecular weight of PolyHSA(60:1) increased the hydrodynamic radius and solution viscosity. Pharmacokinetic analysis of PolyHSA(60:1) indicates reduced clearance and increased circulatory half-life compared with monomeric HSA and other PolyHSA formulations. In conclusion, HSA molecular size and solution viscosity affect central hemodynamics, microvascular blood flow, volume expansion, and circulation persistence during small-volume resuscitation from hemorrhagic shock. In addition, PolyHSA can be an alternative to HSA in pathophysiological situations with compromised vascular permeability.

Nonlinear Cardiovascular Regulation Consequent to Changes in Blood Viscosity

Clinical Hemorheology and Microcirculation. 2011  |  Pubmed ID: 22214675

Increasing blood and plasma viscosity is generally associated with pathological conditions, and increased cardiovascular risk, a perception based in part on studies where blood viscosity is increased to extreme values attained by hemoconcentration. Present studies, supported by epidemiological studies in humans, show that moderate increases in Hct improve cardiovascular function and vice versa. This result is due to the nonlinear regulation of peripheral vascular resistance arising from the increased production of nitric oxide following the increase of shear stress on the vascular wall due to increasing blood viscosity. Similar effects are found in when plasma viscosity is increased in the extremely hemodiluted circulation. In both cases there is an effect at the arteriolar/capillary level, leading to a condition of improved microvascular function and supra perfusion that facilitates clearance of metabolic waste products, while maintaining oxygen delivery. Application of these findings to the design of viscogenic plasma expanders suggests a new approach for the treatment of hemorrhage that in part replaces the use of blood transfusions, making it feasible to lower the transfusion trigger to levels below than normally considered safe.

Improved Resuscitation from Hemorrhagic Shock with Ringer's Lactate with Increased Viscosity in the Hamster Window Chamber Model

The Journal of Trauma. Aug, 2011  |  Pubmed ID: 21248647

Infusion of large volume of fluid is practiced in the treatment of hemorrhagic shock although resuscitation with small fluid volumes reduces the risks associated with fluid overload. We explored the hypothesis that reduced Ringer's lactate (RL) volume restoration in hemorrhage is significantly improved by increasing its viscosity, leading to improved microvascular conditions.

Synthesis and Biophysical Properties of Polymerized Human Serum Albumin

Biotechnology Progress. Jan-Feb, 2011  |  Pubmed ID: 21312376

The use of many plasma expanders (PEs) is often limited by undesirable side effects, such as red blood cell (RBC) aggregation (hydroxyethyl starch), nephrotoxicity (dextran), and extravasation (albumin). Despite its natural prevalence in the bloodstream, human serum albumin (HSA) can increase the risk of mortality when administered to patients with increased vascular permeability (i.e., patients suffering from burns, septic shock, and endothelial dysfunction). The harmful extravasation of HSA can be limited by polymerizing HSA to increase its molecular size. In this study, HSA was nonspecifically cross-linked with glutaraldehyde at different cross-link densities by varying the molar ratio of glutaraldehyde to HSA. The results of this study show that the weight-averaged molecular weight (MW), viscosity, and extent of RBC aggregation of polymerized HSA increases with increasing cross-link density, whereas the colloid osmotic pressure (COP) decreases with increasing cross-link density. Interestingly, circular dichroism measurements indicate that the secondary structure of HSA is unaffected by polymerization. Altogether, these results show that glutaraldehyde can effectively cross-link HSA to produce high MW polymers, yielding a novel series of potential PEs that exhibit low COP and high viscosity.

Delaying Blood Transfusion in Experimental Acute Anemia with a Perfluorocarbon Emulsion

Anesthesiology. Apr, 2011  |  Pubmed ID: 21326091

To avoid unnecessary blood transfusions, physiologic transfusion triggers, rather than exclusively hemoglobin-based transfusion triggers, have been suggested. The objective of this study was to determine systemic and microvascular effects of using a perfluorocarbon-based oxygen carrier (PFCOC) to maintain perfusion and oxygenation during extreme anemia.

Small-volume Resuscitation from Hemorrhagic Shock Using High-molecular-weight Tense-state Polymerized Hemoglobins

The Journal of Trauma. Oct, 2011  |  Pubmed ID: 21336190

The objective of this study was to determine the role of plasma oxygen carrying capacity during resuscitation from hemorrhagic shock (HS).

Exogenous Nitric Oxide Prevents Cardiovascular Collapse During Hemorrhagic Shock

Resuscitation. May, 2011  |  Pubmed ID: 21342744

This study investigated the systemic and microvascular hemodynamic changes related to increased nitric oxide (NO) availability following significant hemorrhage, made available by administration of NO releasing nanoparticles (NO-nps). Hemodynamic responses to hemorrhagic shock were studied in the hamster window chamber. Acute hemorrhage was induced by arterial controlled bleeding of 50% of blood volume, and the resulting hemodynamic parameters were followed over 90 min. Exogenous NO was administered in the form of NO-nps (5mg/kg suspended in 50 μl saline) 10 min following induced hemorrhage. Control groups received equal dose of NO free nanoparticles (Control-nps) and Vehicle solution. Animals treated with NO-nps partially maintained systemic and microvascular function during hypovolemic shock compared to animals treated with Control-nps or the Vehicle (50 μl saline). The continuous NO released by the NO-nps reverted arteriolar vasoconstriction, partially recovered both functional capillary density and microvascular blood flows. Additionally, NO supplementation post hemorrhage prevented cardiac decompensation, and thereby maintained and stabilized the heart rate. Paradoxically, the peripheral vasodilation induced by the NO-nps did not decrease blood pressure, and combined with NO's effects on vascular resistance, NO-nps promoted intravascular pressure redistribution and blood flow, avoiding tissue ischemia. Therefore, by increasing NO availability with NO-nps during hypovolemic shock, it is possible that cardiac stability and microvascular perfusion can be preserved, ultimately increasing survivability and local tissue viability, and reducing hemorrhagic shock sequelae. The relevance, stability, and efficacy of exogenous NO therapy in the form of NO-nps will potentially facilitate the intended use in battlefield and trauma situations.

Nitric Oxide Protection Against Murine Cerebral Malaria is Associated with Improved Cerebral Microcirculatory Physiology

The Journal of Infectious Diseases. May, 2011  |  Pubmed ID: 21415018

Cerebral malaria (CM) is a leading cause of death in Plasmodium falciparum infections. In the Plasmodium berghei ANKA (PbA) murine model, CM pathogenesis is associated with low nitric oxide (NO) bioavailability and brain microcirculatory complications, with a marked decrease in cerebral blood flow, vasoconstriction, vascular plugging by adherent cells, and hemorrhages. Using intravital microscopy through a closed cranial window, here we show that NO supplementation in the form of a NO donor (dipropylenetriamine NONOate [DPTA-NO]) prevented vasoconstriction and improved blood flow in pial vessels of PbA-infected mice. Arterioles and venules of smaller diameters (20-35.5 μm) showed better response to treatment than vessels of larger diameters (36-63 μm). Exogenous NO provided protection against brain hemorrhages (mean, 1.4 vs 24.5 hemorrhagic foci per section) and inflammation (mean, 2.5 vs 10.9 adherent leukocytes per 100 μm vessel length) compared with saline treatment. In conclusion, NO protection against CM is associated with improved brain microcirculatory hemodynamics and decreased vascular pathology.

Integration of Cardiovascular Regulation by the Blood/endothelium Cell-free Layer

Wiley Interdisciplinary Reviews. Systems Biology and Medicine. Jul-Aug, 2011  |  Pubmed ID: 21523919

The cell-free layer (CFL) width separating red blood cells in flowing blood from the endothelial cell membrane is shown to be a regulator of the balance between nitric oxide (NO) production by the endothelium and NO scavenging by blood hemoglobin. The CFL width is determined by hematocrit (Hct) and the vessel wall flow velocity gradient. These factors and blood and plasma viscosity determine vessel wall shear stress which regulates the production of NO in the vascular wall. Mathematical modeling and experimental findings show that vessel wall NO concentration is a strong nonlinear function of Hct and that small Hct variations have comparatively large effects on blood pressure regulation. Furthermore, NO concentration is a regulator of inflammation and oxygen metabolism. Therefore, small, sustained perturbations of Hct may have long-term effects that can promote pro-hypertensive and pro-inflammatory conditions. In this context, Hct and its variability are directly related to vascular tone, peripheral vascular resistance, oxygen transport and delivery, and inflammation. These effects are relevant to the analysis and understanding of blood pressure regulation, as NO bioavailability regulates the contractile state of blood vessels. Furthermore, regulation of the CFL is a direct function of blood composition therefore understanding of its physiology relates to the design and management of fluid resuscitation fluids. From a medical perspective, these studies propose that it should be of clinical interest to note small variations in patient's Hct levels given their importance in modulating the CFL width and therefore NO bioavailability. WIREs Syst Biol Med 2011 3 458-470 DOI: 10.1002/wsbm.150

Synthesis, Biophysical Properties, and Oxygenation Potential of Variable Molecular Weight Glutaraldehyde-polymerized Bovine Hemoglobins with Low and High Oxygen Affinity

Biotechnology Progress. Apr, 2011  |  Pubmed ID: 21584950

In a recent study, ultrahigh molecular weight (M(w) ) glutaraldehyde-polymerized bovine hemoglobins (PolybHbs) were synthesized with low O(2) affinity and exhibited no vasoactivity and a slight degree of hypertension in a 10% top-load model.(1) In this work, we systematically investigated the effect of varying the glutaraldehyde to hemoglobin (G:Hb) molar ratio on the biophysical properties of PolybHb polymerized in either the low or high O(2) affinity state. Our results showed that the M(w) of the resulting PolybHbs increased with increasing G:Hb molar ratio. For low O(2) affinity PolybHbs, increasing the G:Hb molar ratio reduced the O(2) affinity and CO association rate constants in comparison to bovine hemoglobin (bHb). In contrast for high O(2) affinity PolybHbs, increasing the G:Hb molar ratio led to increased O(2) affinity and significantly increased the CO association rate constants compared to unmodified bHb and low O(2) affinity PolybHbs. The methemoglobin level and NO dioxygenation rate constants were insensitive to the G:Hb molar ratio. However, all PolybHbs displayed higher viscosities compared to unmodified bHb and whole blood, which also increased with increasing G:Hb molar ratio. In contrast, the colloid osmotic pressure of PolybHbs decreased with increasing G:Hb molar ratio. To preliminarily evaluate the ability of low and high O(2) affinity PolybHbs to potentially oxygenate tissues in vivo, an O(2) transport model was used to simulate O(2) transport in a hepatic hollow fiber (HF) bioreactor. It was observed that low O(2) affinity PolybHbs oxygenated the bioreactor better than high O(2) affinity PolybHbs. This result points to the suitability of low O(2) affinity PolybHbs for use in tissue engineering and transfusion medicine. Taken together, our results show the quantitative effect of varying the oxygen saturation of bHb and G:Hb molar ratio on the biophysical properties of PolybHbs and their ability to oxygenate a hepatic HF bioreactor. We suggest that the information gained from this study can be used to guide the design of the next generation of hemoglobin-based oxygen carriers (HBOCs) for use in tissue engineering and transfusion medicine applications. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011.

Exogenous Nitric Oxide Decreases Brain Vascular Inflammation, Leakage and Venular Resistance During Plasmodium Berghei ANKA Infection in Mice

Journal of Neuroinflammation. 2011  |  Pubmed ID: 21649904

Cerebral malaria (CM) is a lethal complication of Plasmodium falciparum infections. In the Plasmodium berghei ANKA (PbA) murine model, CM is associated with marked brain inflammation, increased expression of endothelial cell adhesion molecules and leukocyte and platelet accumulation in brain vessels, causing vascular occlusion and decreased blood flow, damaging the endothelium and leading to blood-brain barrier breakdown, leakage and hemorrhages. Exogenous nitric oxide (NO) administration largely prevents the syndrome. Here we evaluated whether the mechanism of action of NO in preventing murine CM is related to its anti-inflammatory properties and to protection of the endothelium.

Nanoengineering Bactericidal Nitric Oxide Therapies

Virulence. May-Jun, 2011  |  Pubmed ID: 21670586

Exogenous Intravascular Nitric Oxide Enhances Ventricular Function After Hemodilution with Plasma Expander

Life Sciences. Jan, 2012  |  Pubmed ID: 22056371

This study evaluated the hypothesis that exogenous nitric oxide (NO) supplementation during acute hemodilution with plasma expander (PE) provides beneficial effects on cardiac function.

Effects of Acute Anemia and Hyperoxia on Oxygen Distribution

Advances in Experimental Medicine and Biology. 2012  |  Pubmed ID: 22259104

Simulation of NO and O(2) Transport Facilitated by Polymerized Hemoglobin Solutions in an Arteriole That Takes into Account Wall Shear Stress-induced NO Production

Biophysical Chemistry. Jan, 2012  |  Pubmed ID: 22285312

A mathematical model was developed to study nitric oxide (NO) and oxygen (O(2)) transport in an arteriole and surrounding tissues exposed to a mixture of red blood cells (RBCs) and hemoglobin (Hb)-based O(2) carriers (HBOCs). A unique feature of this model is the inclusion of blood vessel wall shear stress-induced production of endothelial-derived NO, which is very sensitive to the viscosity of the RBC and HBOC mixture traversing the blood vessel lumen. Therefore in this study, a series of polymerized bovine Hb (PolyHb) solutions with high viscosity, varying O(2) affinities, NO dioxygenation rate constants and O(2) dissociation rate constants that were previously synthesized and characterized by our group was evaluated via mathematical modeling, in order to investigate the effect of these biophysical properties on the transport of NO and O(2) in an arteriole and its surrounding tissues subjected to anemia with the commercial HBOC Oxyglobin® and cell-free bovine Hb (bHb) serving as appropriate controls. The computer simulation results indicated that transfusion of high viscosity PolyHb solutions promoted blood vessel wall shear stress dependent generation of the vasodilator NO, especially in the blood vessel wall and should transport enough NO inside the smooth muscle layer to activate vasodilation compared to the commercial HBOC Oxyglobin® and cell-free bHb. However, NO scavenging in the arteriole lumen was unavoidable due to the intrinsic high NO dioxygenation rate constant of the HBOCs being studied. This study also observed that all PolyHbs could potentially improve tissue oxygenation under hypoxic conditions, while low O(2) affinity PolyHbs were more effective in oxygenating tissues under normoxic conditions compared with high O(2) affinity PolyHbs. In addition, all ultrahigh molecular weight PolyHbs displayed higher O(2) transfer rates than the commercial HBOC Oxyglobin® and cell-free bHb. Therefore, these results suggest that ultrahigh molecular weight PolyHb solutions could be used as safe and efficacious O(2) carriers for use in transfusion medicine. It also suggests that future generations of PolyHb solutions should possess lower NO dioxygenation reaction rate constants in order to reduce NO scavenging, while maintaining high solution viscosity to take advantage of wall shear stress-induced NO production. Taken together, we suggest that this mathematical model can be used to predict the vasoactivity of HBOCs and help guide the design and optimization of the next generation of HBOCs for use in transfusion medicine.

Hypervolemic Infusion of Lumbricus Terrestris Erythrocruorin Purified by Tangential-flow Filtration

Transfusion. Feb, 2012  |  Pubmed ID: 22304397

BACKGROUND: The hemoglobin of the earthworm Lumbricus terrestris (also known as erythrocruorin, or LtEc) is a naturally occurring high-molecular-weight protein assembly (3.6 MDa) that is extremely stable, resistant to oxidation, and transports oxygen similarly to human whole blood. Therefore, LtEc may serve as an alternative to donated human red blood cells. However, a suitable purification process must be developed to produce highly pure LtEc on a large scale that can be evaluated in an animal model to determine the safety and efficacy of LtEc. STUDY DESIGN AND METHODS: We used tangential-flow filtration (TFF), an easily scalable and affordable technique, to produce highly pure LtEc from earthworms. The purity, yield, methemoglobin level, viscosity, colloid osmotic pressure, O(2) binding equilibria, and ligand-binding kinetics of the purified LtEc were measured in vitro. The purified LtEc product was then evaluated in hamsters using a hypervolemic infusion model to establish its basic biocompatibility and detect any changes in microcirculatory and systemic variables. RESULTS: TFF was able to produce LtEc with high purity and yield (5-10 g/1000 worms). The purified LtEc product did not elicit hypertension or vasoconstriction when infused into hamsters. CONCLUSION: LtEc may be easily purified and safely transfused into hamsters in small amounts (0.5-1.5 g/dL final concentration in blood) without any noticeable side effects. Therefore, LtEc may serve as a very promising oxygen carrier for use in transfusion medicine.