Bovine Hemoglobin Solution Research Articles

A novel hemoglobin-based blood substitute protects against myocardial reperfusion injury

HBOC-201 (Biopure; Cambridge, MA) is a glutaraldehyde-polymerized bovine hemoglobin (Hb) solution that is stroma free, has lower viscosity than blood, and promotes O(2) unloading. We investigated the effects of HBOC-201 in a canine model of myocardial ischemia-reperfusion injury. Dogs were anesthetized and subjected to 90 min of regional myocardial ischemia and 270 min of reperfusion. HBOC-201 or 0.9% saline vehicle equivalent to 10% total blood volume was infused 30 min before myocardial ischemia. Hemodynamic data and peripheral blood samples were taken at baseline, 1 h of myocardial ischemia, and 1, 2, and 4 h of reperfusion. At 270 min of reperfusion, the area at risk (AAR) per left ventricle and the area of infarction (Inf) per AAR were determined. The myocardial AARs in the two study groups were similar. In addition, myocardial blood flow (as measured by radioactive microspheres) in the ischemic zone was similar between the vehicle and HBOC-201 groups. HBOC-201-infused dogs demonstrated a significant (P < 0.01) 56% reduction in Inf/AAR. Analysis of blood samples taken at 4 h of reperfusion showed a significant (P < 0.05) reduction in creatine kinase MB isoform for the HBOC-201 group. Histological analysis of the myocardium demonstrated significant (P < 0.01) reductions in neutrophil infiltration in the HBOC-201 group. These data indicate that treatment with HBOC-201 before myocardial ischemia-reperfusion reduces the extent of myocardial inflammation and ischemia-reperfusion injury in the canine myocardium.

New generation of hemoglobin-based oxygen carriers evaluated for oxygenation of critically ischemic hamster flap tissue

The aim of this study was to investigate and compare the effects of a traditionally formulated, low-viscosity, right-shifted polymerized bovine hemoglobin solution and a highly viscous, left-shifted hemoglobin vesicle solution (HbV-HES) on the oxygenation of critically ischemic peripheral tissue. Randomized, prospective study. University laboratory. A total of 40 male golden Syrian hamsters. Island flaps were dissected from the back skin of anesthetized hamsters. The flap included a critically ischemic, hypoxic area that was perfused via a collateralized vasculature. One hour after completion of the preparation, the animals received a 33% blood exchange with 6% hydroxyethyl starch 200/0.5 (HES, n = 9), HbV suspended in HES (HbV-HES, n = 8), or polymerized bovine hemoglobin solution (n = 9). Three hours after the blood exchange, microcirculatory blood flow (laser-Doppler flowmetry) was increased to 262% of baseline for HbV-HES (p < .01) and 197% for polymerized bovine hemoglobin solution (p < .05 vs. baseline and HbV-HES). Partial tissue oxygen tension (bare fiber probes) was only improved after HbV-HES (9.4 torr to 14.2 torr, p < .01 vs. baseline and other groups). The tissue lactate/pyruvate ratio (microdialysis) was elevated to 51 in the untreated control animals, and to 34 +/- 8 after HbV-HES (p < .05 vs. control) and 38 +/- 11 after polymerized bovine hemoglobin solution (not significant). Our study suggests that in critically ischemic and hypoxic collateralized peripheral tissue, oxygenation may be improved by normovolemic hemodilution with HbV-HES. We attributed this improvement to a better restoration of the microcirculation and oxygen delivery due to the formulation of the solution.

Influence of the Hemoglobin Solution HBOC-201 on Tissue Oxygenation in the Rat R1H-Tumor

HBOC-201 is an ultra purified bovine hemoglobin solution. It has already been used in clinical phase II/III trials for emergency treatments. Animal experiments have shown that HBOC-201 is highly effective in tissue oxygenation. The study was performed in order to assess the potential of low dose HBOC-201 to improve tumor oxygenation. 30 rats with a subcutaneously growing rhabdomyosarcoma R1H tumor were randomly assigned either to be ventilated with carbogen (n = 10), or to receive an IV injection of 0.3 g/kg HBOC-201 (n = 10) or a combination of 0.3 g/kg HBOC-201 and carbogen breathing (n = 10). Under general anesthesia the effects of the respective treatment on the tissue oxygen tension (tpO2) of the tumor were determined using a flexible stationary probe at baseline (b) and 15 and 60 min after application of the respective medication. HBOC-201 alone failed to improve tumor tpO2 (b: 1.3 +/- 1.2mmHg; 15min: 1.4 +/- 1 mmHg; 60min: 1 +/- 1 mmHg). In combination with carbogen the mean tpO2 of the tumor raised in comparison to baseline values (b: 3.1 +/- 4.6 mmHg; mmHg; 15min: 8.5 +/- 11*mmHg; 60min: 4.8 +/- 5mmHg; *p < 0.05 vs. b), but this effect was less pronounced than the increase in tpO2 by carbogen alone (b: 3.4 +/- 3.4mmHg; 15min: 9 +/- 10* mmHg; 60 min: 13 +/- 19* mmHg; *p < 0.05 vs. b). The application of low dose hemoglobin solution HBOC-201 does not result in improvement of tissue oxygenation in the rat rhabdomyosarcoma R1H.

Application of the bovine hemoglobin solution HBOC-301 with and without isovolemic hemodilution reduces the histopathologic damage in acute severe pancreatitis in swine

Severe acute pancreatitis (AP) remains associated with a high mortality. In experimental AP the application of the bovine hemoglobin solution HBOC-301 with and without IHD was able to normalize pancreatic tissue oxygen tension [1,2]. The aim of this study was to evaluate the therapeutic approach of volume therapy with HES plus HBOC-301 with and without IHD with regard to the histologic damage and survival time in pigs suffering from AP.

Growth and antioxidant status of plant cells cultured with bovine haemoglobin solution.

Reactive oxygen species (ROS), such as superoxide (O2 •-), hydrogen peroxide (H2O2), and the hydroxyl radical (•OH), may be generated in plant cells during aerobic metabolism. ROS can impair cell activities, primarily through oxidative damage to lipids, proteins and nucleic acids (Halliwell and Gutteridge, 1999). Exposure of plants or their cells/tissues, maintained in vitro, to environmental stresses (e.g. temperature extremes, drought, high salinity, mineral deficiency) perturbs the balance between the ROS production and the quenching effects of antioxidant enzymes, leading to oxidative damage (Smirnoff, 1993; Foyer and Mullineaux, 1994; Schwanz et al., 1996). However, plants/cells possessing high activities of constitutive or induced antioxidant enzymes show increased resistance to such oxidative damage (Halliwell and Gutteridge, 1999; Niki, 2000).KeywordsAntioxidant StatusMitotic DivisionCallus Induction MediumCellular Reactive Oxygen SpeciesPlant Cell GrowthThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Polymerized bovine hemoglobin solution as a replacement for allogeneic red blood cell transfusion after cardiac surgery: Results of a randomized, doubleblind trialJ. Levy, L. Goodnough, P.E. Greilich, et al. J. Thorac Cardiovasc Surg 124:35–42, 2002

Polymerized bovine hemoglobin solution as a replacement for allogeneic red blood cell transfusion after cardiac surgery: Results of a randomized, doubleblind trialJ. Levy, L. Goodnough, P.E. Greilich, et al. J. Thorac Cardiovasc Surg 124:35–42, 2002

The use of ultrapurified bovine hemoglobin solution in the treatment of two cases of presumed red maple toxicosis in a miniature horse and a pony

AbstractObjective: To report the use of ultrapurified bovine hemoglobin (PBHg) in the treatment of red maple toxicosis in horses.Summary: The use of PBHg in a miniature horse and a pony with clinical signs of red maple toxicosis is described. Initial treatment of the horses included blood transfusion in Case 1, and intravenous crystalloid fluids, nasogastrically administered mineral oil, and activated charcoal in both cases. Both cases received PBHg (16 and 11 ml/kg, respectively) when the hematocrit dropped below 10%. Clinical stabilization of the horses (reflected by improved vital signs and mucous membrane color, increase in PvO2, and increase in plasma hemoglobin) was temporary, lasting 12 and 42 hours, respectively. A whole blood transfusion was given 16 hours after the infusion of PBHg in Case 1 and 42 hours after the infusion of PBHg in Case 2. The horses were discharged on days 15 and 17, respectively. Both were stable on discharge and were reported to be doing well 1 month after discharge. New information provided: Ultrapurified bovine hemoglobin is shown to be safe in the treatment of red maple toxicosis. Ultrapurified bovine hemoglobin infusion resulted in temporary improvement in oxygen (O2) delivery as assessed by clinical and laboratory parameters. The use of PBHg may provide an ‘oxygen bridge’ in an acute hemolytic crisis in the horse. It may allow additional time in the treatment of red maple toxicity for the acute hemolysis to subside and for metabolism of the toxin to occur, thus providing a more optimal environment for an effective whole blood transfusion.

Polymerized bovine hemoglobin solution as a replacement for allogeneic red blood cell transfusion after cardiac surgery: Results of a randomized, double-blind trial

Background: Blood loss leading to reduced oxygen-carrying capacity is usually treated with red blood cell transfusions. This study examined the hypothesis that a hemoglobin-based oxygen-carrying solution can serve as an initial alternative to red blood cell transfusion. Methods: In a randomized, double-blind efficacy trial of HBOC-201, a total of 98 patients undergoing cardiac surgery and requiring transfusion were randomly assigned to receive either red blood cell units or HBOC-201 (Hemopure; Biopure Corporation, Cambridge, Mass) for the first three postoperative transfusions. Patients were monitored before and after transfusion, at discharge, and at 3 to 4 weeks after the operation for subsequent red blood cell use, hemodynamics, and clinical laboratory parameters. Results: The use of HBOC-201 eliminated the need for red blood cell transfusions in 34% of cases (95% confidence interval 21%-49%). Patients in the HBOC group received a mean of 1.72 subsequent units of red blood cells; those who received red blood cells only received a mean of 2.19 subsequent units (P =.05). Hematocrit values were transiently lower in the HBOC group but were similar in the two groups at discharge and follow-up. Oxygen extraction was greater in the HBOC group (P =.05). Mean increases in blood pressure were greater in the HBOC group, but not significantly so. Conclusion: HBOC-201 may be an initial alternative to red blood cell transfusions for patients with moderate anemia after cardiac surgery. In a third of cases, HBOC-201 eliminated the need for red blood cell transfusion, although substantial doses were needed to produce this modest degree of blood conservation.J Thorac Cardiovasc Surg 2002;124:35-42

Preservation of myocardial energy status by bovine hemoglobin solutions during ischemia

Compared to murine and human hemoglobin, bovine hemoglobin has a less exothermic oxygen binding and delivers oxygen even at low temperatures. This property could improve oxygen availability for myocytes during hypothermic arrest of hearts. The aim of this study was to evaluate the advantage of using cardioplegic solutions enriched with bovine hemoglobin when storing rat hearts. Hearts excised from rats after perfusion with different cardioplegic solutions (Celsior, Celsior plus 4% human hemoglobin, Celsior plus 4% and 8% bovine hemoglobin) were compared. Biopsies were obtained from the beating hearts before cardioplegic infusion and during a 48 h period of cold storage. Adenosine triphosphate, its catabolites and markers of oxidative stress were measured as indices of preservation. The results show that bovine hemoglobin-enriched solutions highly improve adenosine triphosphate content, decreasing its catabolites; no significant changes in antioxidant status were evident. The statistically significant difference was evident up to 6 h of storage. Doubling the concentration of bovine hemoglobin produces only slight improvement. Alternative hemoglobins with different properties may improve and prolong heart storage. As bovine hemoglobin delivers oxygen even at low temperatures, it improves energy content and anabolic reactions, without decreasing oxidative stress.

Effects of Ultra-Purified Polymerized Bovine Hemoglobin on Local Tissue Oxygen Tension in Striated Skin Muscle – An Efficacy Study in the Hamster

The development of hemoglobin-based oxygen carriers has been propagated for replacement of the oxygen carrying properties of red blood cells for almost one century. Using a Clark-type multi-wire oxygen surface electrode and the dorsal skin fold chamber model of the awake Syrian golden hamster, local tissue pO₂ was analyzed in the thin striated skin muscle before and after administration of an ultrapurified polymerized bovine hemoglobin solution (U-PBHb^®, Biopure Corp., Boston, Mass., USA) under the following experimental conditions: (a) hypervolemic infusion with U-PBHb at ∼10% of calculated blood volume, and (b) isovolemic exchange transfusion with U-PBHb by replacing ∼50% of calculated blood volume. Control animals of group a received equivalent treatment with either isotonic saline or dextran 60, control animals of group b received dextran 60. Local tissue pO₂ was found slightly decreased after both hypervolemic infusion and isovolemic exchange transfusion with U-PBHb, while frequency distribution curves of local tissue pO₂ were found more narrow (less values <10 mm Hg and >25 mm Hg), suggesting a more homogeneous tissue pO₂ distribution. The data thus indicate that U-PBHb slightly decreases mean tissue pO₂ after both hypervolemic infusion and isovolemic exchange transfusion which is accompanied by an effective homogenization of local tissue pO₂ distribution as compared to dextran 60.

Current status of cell-free hemoglobin solutions

Current Status of Cell-Free Haemoglobin Solutions. Persisting concern about the risks of allogeneic blood and the impending shortfall of such products have reinforced the search for alternative erythrocyte substitutes in recent years. With the absence of toxic side effects of modern, ultrapurified and chemically modified haemoglobin-based oxygen carriers (HBOC), many studies in animals, volunteers and patients have shown their effectivity and tolerability. HBOC can be applied in patients without prior cross-matching and one product, which is available as room temperature-stable formulation with a shelf-life of > 2 years, has recently been approved for the treatment of peri-operative anaemia in South Africa. While clinical studies with human cross-linked haemoglobin (DCLHb) were discontinued because of an increased mortality of patients receiving DCLHb after stroke and multiple injury shock, phase III studies with polymerized bovine haemoglobin (HBOC-201) have been completed showing that peri-operative infusion of HBOC-201 was able to avoid or reduce allogeneic blood transfusion. Different human, recombinant and bovine hemoglobin solutions are currently under investigation in cardiac surgery, sepsis and tumour patients. Extensive laboratory investigations have shown that HBOC are highly effective oxygen donors with regard to tissue oxygenation rather than erythrocyte substitutes in case of bleeding, because they provide plasmatic oxygen delivery under physiological concentrations of inspired oxygen. HBOC have advantages over red blood cells because of their easier oxygen release per se and from adjacent erythrocytes, and their capability to reach poststenotic or poorly perfused tissues with the plasma stream.

Hemoglobin-stimulated growth and antioxidant activities in cultured cotton cells

Changes in cellular reactive oxygen scavenging enzymes were assessed in suspension-derived cells of cotton (Gossypium herbaceum) cv. Dhumad following culture with a commercial bovine hemoglobin (Hb) solution (Erythrogen) at 1:100–1:1000 (v:v). Mean (± SEM) fresh (f.wt.) and dry weights (d.wt.) of cells after 25 d of culture were significantly (p < .05) greater in medium supplemented with 1:750 and 1:1000 (v:v) Erythrogen, compared to controls lacking Erythrogen. For example, with 1:750 (v:v) Erythrogen, mean cell f.wt. and d.wt. were increased by 45 and 31%, respectively. Total soluble cellular protein increased by 141, 176, and 191% with Erythrogen at 1:50, 1:750, and 1:1000 (v:v), respectively. Cellular catalase and glutathione reductase activities decreased significantly (p < .05) following addition of low concentrations (1:1000 and 1:750 v:v) of Erythrogen to culture medium. However, increasing the concentration of Erythrogen to a maximum of 1:100 (v:v), caused a concomitant increase in catalase to a maximum of 62% over control. Mean total superoxide dismutase activity increased linearly with increasing Erythrogen concentration, reaching a maximum mean value over 2-fold greater than control with 1:100 (v:v) Erythrogen. A similar trend was observed in cellular H2O2 content, which reached a maximum of 98% over control with 1:250 (v:v) Erythrogen. These results demonstrate that culture of cotton cells with Hb solution causes changes in cellular oxygenation sufficient to modify cellular antioxidant status.

Bovine haemoglobin solution and gelatin infusion in a rabbit model of arterial thrombosis and bleeding

Bovine haemoglobin solution and gelatin infusion in a rabbit model of arterial thrombosis and bleeding

Microspore-derived embryo induction from cultured anthers of wheat.

Microspore-derived embryo (MDE) induction was evaluated following the culture of wheat (Triticum aestivum L.) anthers in a basal medium with a range of supplements. Several supplements were evaluated, including a commercial bovine haemoglobin solution (Erythrogen™), Ficoll and the co-polymer surfactant Pluronic F-68, but these did not enhance MDE induction. However, phenylacetic acid, replacing both 2,4-dichlorophenoxyacetic acid and kinetin, increased MDE induction by a factor of 1.6 (p<0.05), producing 257±29 (mean±s.e., n=3) MDEs per 100 anthers cultured. Twenty-four plants regenerated from MDEs induced via anther culture were used for ploidy determinations; 33% were sterile haploids, while the remainder were fertile.

Haemoglobin (Erythrogen)-enhanced post-thaw growth of cryopreserved cells.

Supplementation of semi-solid R2 culture medium with a commercial bovine haemoglobin (Hb) solution (Erythrogen) at 1:50-1:500 (v:v), had beneficial effects on the growth, following cryopreservation, of cells of the Indica rice, Oryza sativa cv. Pusa Basmati 1. The mean absorbance, as assessed by triphenyl tetrazolium chloride reduction, of rice cells at 8 d post-thawing, was increased by up to 60% (P < 0.05), compared to cells recovered in the absence of Hb. Erythrogen (1:50-1:500 v:v) promoted an increase in biomass, of up to 25% over control (P < 0.05), at 24 d post-thawing. Cell suspensions, re-established by transfer to liquid medium of cells initially thawed and cultured with Erythrogen for 24 d, exhibited increased (up to 2-fold) growth rates over a subsequent 20-d period, compared to cells recovered without Hb.

Administration of polymerized bovine hemoglobin improves survival in a rat model of traumatic shock.

The ability of purified and modified hemoglobin solutions to expand blood volume and act as oxygen and carbon dioxide transporters with no apparent toxic side effects has lead to an increased interest in their utilization in shock conditions. Although hemoglobin solutions have been shown to provide beneficial effects in endotoxic and hemorrhagic shock, they have not been studied in whole body traumatic injury. We investigated the effects of a polymerized bovine hemoglobin solution (HBOC-201) in a rat model of traumatic shock. Pentobarbital-anesthetized rats subjected to Noble-Collip drum trauma developed a shock state characterized by marked hypotension, a survival time of 102 +/- 20 min, significant increases in intestinal myeloperoxidase (MPO) activity and splanchnic vascular endothelial dysfunction characterized by an impaired vasorelaxation of the superior mesenteric artery (SMA) to endothelium-dependent vasodilators. Traumatized rats were treated with HBOC-201 or an equal volume of vehicle corresponding to 5, 10 or 15% of the calculated blood volume. Treatment with HBOC-201 10 min posttrauma prolonged survival, normalized mean arterial blood pressure and attenuated endothelial dysfunction of the SMA. However, administration of HBOC-201 failed to significantly attenuate increases in intestinal MPO activity following trauma. No beneficial effects were observed with the administration the vehicle. These data indicate that HBOC-201 exerts significant beneficial effects in traumatic shock states by normalizing systemic blood pressure and attenuating vascular endothelial dysfunction. HBOC-201 may serve as a useful adjunctive agent in the early treatment of trauma.

Lyophilized bovine hemoglobin as a possible reference material for the determination of hemoglobin derivatives in human blood

We investigated the suitability of a lyophilized bovine hemoglobin (LBH) preparation containing various fractions of oxyhemoglobin (O2Hb), carboxyhemoglobin (COHb), and methemoglobin (MetHb) for quality assessment in multicomponent analysis (MCA) of hemoglobin derivatives. It was demonstrated that a stable preparation of these components after reconstitution yields a hemoglobin solution that is spectrophotometrically equivalent with a fresh bovine hemoglobin solution. The preparation was found to be stable for at least 1 year when it is kept at 2-8 degrees C and for 1 h after reconstitution. We determined the fractions of O2Hb, COHb, and MetHb of several LBH preparations, using the complete spectra of 480-650 nm with 2-nm intervals and absorptivities as determined for pure LBH solutions. A field trial involving various types of multiwavelength hemoglobin photometers showed the suitability of LBH as a quality-control material. Computer models of the various common multiwavelength hemoglobin photometers may be useful for establishing more accurate target values of LBH preparations for each type of photometer and for studying the importance of the influence of specific factors such as wavelength selection, absorptivity values, and interfering dyes.

Acoustic properties of egg yolk and albumen in the frequency range 20-400 MHz.

The acoustic propagation properties of egg yolk and albumen are characterized in the frequency range 20-400 MHz by the bioultrasonic spectroscopy system using an ultrasonic transmission comparison method. Significant differences in the attenuation, velocity, impedance, and density among yolk and thick and outer thin albumen are observed. The acoustic properties of 10% aqueous solutions of ovalbumin and bovine hemoglobin are also measured in order to investigate the contribution of proteins to the acoustic properties of albumen. The differences obtained between thick and outer thin albumen may be mainly due to their macromolecular level structural differences, as their constituents are nearly the same.

Effect of Hemopure® on Prothrombin Time and Activated Partial Thromboplastin Time on Seven Coagulation Analyzers,

Hemopure®, a newly developed hemoglobin-based oxygen carrying solution (HBOC-201), is currently undergoing phase II clinical trials in elective surgical patients. Hemopure is a glutaraldehyde-polymerized, stroma free, bovine hemoglobin solution that is administered intravenously. Patients infused with this solution showed highly increased concentrations of plasma hemoglobin as expected. Since plasma hemoglobin may affect the accuracy of many instruments and assays (1), we studied the effect of Hemopure on the performance of commonly used coagulation analyzers. This study was conducted on seven different instrument models from four manufacturers and covered four types of methodologies that are widely available. The assays targeted were prothrombin time (PT) and activated partial thromboplastin time …

Effect of Hemoglobin-Based Oxygen Carrier-201 on Common Chemistry Laboratory Procedures

Hemoglobin-based oxygen carrier-201 (HBOC) contains a solution of glutaraldehyde-polymerized bovine hemoglobin. The hemoglobin molecule absorbs light at several wavelengths. The presence of HBOC increases plasma and serum hemoglobin, producing the appearance of hemolysis. Young lists >80 analytes for which clinical laboratory tests are affected by hemolysis [1] . In preparation for a multicenter study to evaluate the effect of the use and safety of HBOC (Biopure Corp.), we studied the effect of this drug on most of our instruments and analytes that might be used in management of our patients during the planned clinical trial. The instruments studied were: …