Red Blood Cell Solutions Research Articles

Appropriate transport of whole blood by drones for cases of massive bleeding

Background: Whole blood (WB) can be superior to red blood cell (RBC) solutions for transfusion in hemorrhage because it contains coagulation factors and platelets. A previous study reported that the quality of an RBC solution and WB can be maintained during transport by an uncrewed aerial vehicle (drone) at normal temperature. Therefore, we aimed to determine whether this approach could also be used for transport of WB for cases of massive bleeding on the hot and cold days that occur in Japan, by testing this approach twice, on hot and cold days. Method: We used an automobile-transported WB packs obtained by phlebotomy and corresponding segments to a test place in Tomi at a distance of approximately 200 km from Tokyo. Subsequently, a drone (M1000) flight transported the WB and its segments. Results: The influence of the drone flight on WB appeared to be minimal on lactate dehydrogenase (LDH) level, and the WB was confirmed to be problem-free in coagulation tests. Conclusion: We transported WB by the drone and confirmed that there were little hemolysis of the WBs and little reduction in the levels of coagulants.

Effects of transport using an unmanned aerial vehicle on blood samples for cross-matching tests

Objective: This study aimed to examine the effects of transport using an unmanned aerial vehicle (UAV) on blood samples for cross-matching tests in the transfusion of red blood cell solution. Materials and methods: Two bags of red blood cell (RBC) solution (blood type, A, RBC#1 and #2; 280 mL per bag) were obtained from the Japan Red Cross Society. The RBC solution in each bag was further separated into three bags (93 mL per bag). The UAV group consisted of RBC solution with a segment tube containing the whole blood in a temperature controlled cooler box transported using an UAV. The control group consisted of RBC solution with a segment tube containing the whole blood in an in-house refrigerator that was not transported. Samples from the RBC solution were collected on day 10 (2 days before transport) and day 13 (day after transport), and lactate dehydrogenase (LD), potassium, blood sugar, and lactic acid levels were measured. After transport, cross-matching tests were performed using whole blood in the segment tubes and patient`s blood (blood type, type A). Results: We compared the biochemistry between the UAV and control group and found no significant differences, except for LD level. Although the LD level in the UAV group slightly increased compared with that in the control group, the whole blood containing the segment tubes was not hemolyzed, and cross-matching tests were performed. Conclusion: We could fly and transport an RBC solution with segment tubes using an UAV, and this mode of transport did not affect the cross-matching tests, despite mild hemolysis in the RBC solutions.

An active transport refrigerator is optimal for blood preservation in small medical facilities

Objective: In small medical institutions in Japan, red blood cell (RBC) solutions are commonly stored in household refrigerators because of the high cost of blood-only refrigerators. Therefore, we assessed and compared the methods used for preserving the RBC solutions in an active transport refrigerator (ATR), a blood-only refrigerator, and a household refrigerator. Materials and Methods: Irradiated RBC solution samples (280 mL) supplied by the Japanese Red Cross Society (experiments 1–8) and in-house whole blood waste (experiments 9–10) were each divided into two bags. The quality of blood stored in an active transport refrigerator was compared with that of blood stored in a blood-only refrigerator or a household refrigerator. We tested the hemolytic effects of storage and filtration through a transfusion set on the lactate dehydrogenase ratio (LDR) of the RBC solutions. Results: Although the LDR of the RBC solutions stored in the ATR was similar to that of the RBC solutions stored in the blood-only refrigerator, the LDR of the RBC solutions stored in the household refrigerator was significantly higher than that of the RBC solutions stored in the ATR. Conclusion: Temperature management using an ATR is critical for the maintenance of the quality of the RBC solution, which is not achieved by a household refrigerator.

Antagonistic effect of the Fengzhecao extract against human red blood cell hemolysis induced by wasp venom

To explore the antagonistic effect of Fengzhecao extract against human red blood cell (RBC) hemolysis induced by wasp venom. Water extract method was used to extract dried Fengzhecao and vacuum-dried to obtain Fengzhecao extract. It was diluted into 1 g/L for next use. Wasp venom was collected from the wasp workers. A, B, O, AB type healthy blood donors' suspended RBC solution was obtain to make washed RBC solutions and adjust the RBCs count (4.0-80.0)×109/L (the number of RBC counted on the hemocytometer is 1-20 cells/small checker). According to treatment factors, they were divided into the normal saline controlled group (NS group; 200 μL RBC solution+20 μL normal saline), Fengzhecao extract group (FZC group; 200 μL RBC solution+10 μL Fengzhecao extract+10 μL normal saline), wasp venom group (FD group; 200 μL RBC solution+10 μL wasp venom+10 μL normal saline), and Fengzhecao extract+wasp venom group (FCD group; 200 μL RBC solution+10 μL Fengzhecao extract+10 μL wasp venom), with 10 blood samples per group of every blood type. The solutions were put into the glass test tube respectively, and then into 37 centigrade water bath thermostat. After 10 minutes, the blood cell counting plate was directly observed under the microscope and the RBCs was counted. Differences in RBC count was compared between the same treatment factors of different blood types and between different treatment factor groups of the same blood type. There was no statistically significant difference in RBC count between blood types under the same treatment factors. The RBC count (×109/L) of the type A, B, O, AB in the NS group were 5.567±1.368, 5.146±1.690, 4.577±0.774, 5.197±1.587 (F = 0.852, P = 0.475), the FZC group were 5.751±1.489, 5.268±1.418, 4.727±1.174, 5.298±1.229 (F = 0.987, P = 0.410), the FD group were 0.546±0.450, 0.804±0.428, 0.679±0.283, 0.846±0.453 (F = 1.089, P = 0.366), and the FCD group were 5.532±1.330, 5.051±1.596, 4.589±0.879, 5.140±1.492 (F = 0.820, P = 0.492), respectively. Comparison of RBC count between groups with different treatment factors of the same blood type was done. There was no significant difference between the FZC group and the NS group, indicating that the extract of Fengzhecao extract had no effect on hemolysis of RBC; in the FD group, it was significantly lower than the NS group (all P < 0.05), indicating that wasp venom had a significant hemolytic effect on RBC; but there was no statistically significant difference in RBC count between the FCD group and the NS group, indicating that the Fengzhecao extract antagonizes the hemolytic effect of wasp venom without affecting the RBC count; however, the RBC count in the FCD group was significantly higher than that in the FD group (all P < 0.05), further indicating that the Fengzhecao extract antagonizes the hemolytic effect of wasp venom. Wasp venom has a significant hemolytic effect which can be effectively antagonized by Fengzhecao extract and has nothing to do with the human ABO blood type.

The Quality of Blood is not Affected by Drone Transport: An Evidential Study of the Unmanned Aerial Vehicle Conveyance of Transfusion Material in Japan

Unmanned aerial vehicles (UAVs), or drones, are used in Rwanda for transfusion transport, but they have not yet been used in Japan. This technology holds promise for transporting medical supplies during disasters or to remote places where the terrain makes it difficult to travel by land. One of the difficulties in using UAVs is the temperature-control requirements for red blood cell (RBC) solutions, i.e., 2 °C to 6 °C according to Japanese regulations. This study aimed to describe the effectiveness of UAV-based transport of RBC solution. For testing, we gradually increased the UAV travel distance, monitored the temperature of the RBC solution, and conducted laboratory tests to check the integrity of the blood sample. Lactate dehydrogenase (LD) was used as a hemolytic index to indicate the effect of the UAV flight on the blood samples. The UAV was able to exceed 7 km of travel distance despite the relatively heavy load needed for the RBC solution storage. The LD level was not significantly different between the flight and non-flight (control) samples. However, we were not able to completely maintain a temperature of 2 °C to 6 °C; nonetheless, the deviation was within the safe range.

Temperature Management of Red Blood Cell Solution Transported by Car for Transfusion at Home

Objective: In Japan, red blood cell (RBC) solution is usually transported by car from a medical institution to the patient’s house for home transfusion. However, there are no regulations for transporting blood by car in the medical setting. Therefore, we assessed and compared the methods (containers) used for transporting the RBC solution by car. Materials and Methods: Irradiated RBC solution samples (280 mL) supplied by the Japan Red Cross Society were each divided into two bags. The quality of blood transported by car (1 - 2 hours) in an active transport refrigerator (ATR) (control group) was compared with that transported in a cooler, or styrofoam box (study group). We tested the hemolytic effects of transportation by car, storage, and filtration through a transfusion set on the lactate dehydrogenase (LD) levels in the RBC solutions. Results: Post-filtered LD levels were significantly higher in the RBC solutions transported in a cooler-box with inadequate temperature control when compared to those transported in an ATR with optimal temperature control. However, under conditions of optimal temperature control, the post-filtered LD levels were comparable in the control and study (both cooler and styrofoam boxes) group RBC solutions. Conclusion: Temperature management is critical for the maintenance of the quality of the RBC solution transported by car.

The quality of blood dropped from an unmanned aerial vehicle (Drone)

Objective: To evaluate the risk of deterioration in the quality of blood transported by a fixed-wing drone. Materials and methods: Red blood cell (RBC) solutions were categorized into “travel group” (transported by train or car to test place) and “non-travel” group. The travel group was further subcategorized into “dropped,” and “non-dropped” groups. One bag each from the dropped and non-dropped groups were wrapped in an Absorption Sheet (A-Sheet) and an Under Sheet (U-Sheet). RBC solution of dropped group was dropped onto the ground from an altitude of 10 Lactate dehydrogenase (LD) levels from the samples of RBC solution were measured and the LD ratio (LDR) was calculated as the rate of increase of the LD which can be compared. We compared the differences in LDR between the dropped and non-dropped groups. Results: In the dropped group, there was an increase in LDR before filtration in the blood wrapped in the A-sheet. No increase was noted in the blood wrapped in the U-sheet. LDR levels after filtration were slightly increased in the blood samples wrapped in the U-sheet and clearly increased in blood samples wrapped in the A-Sheet. Conclusion: We considered the possibility of blood getting hemolyzed due to a fall and observed that hemolysis was affected by packing. Thus, since it may be difficult to find good packaging for blood, dropped blood may not be safe for transfusion due to the likelihood of hemolysis. Hence, for practical purposes, using a drone without a fixed-wing may be preferable for transportation of blood.

Red blood cell storage and adhesion to vascular endothelium under normal or stress conditions: An in vitro microfluidic study.

Observational studies have identified an association between duration of red blood cell (RBC) storage and adverse outcomes in trauma. Hemorrhagic shock (HS) leads to impaired tissue perfusion which is associated with endothelial cell glycocalyx (eGC) shedding. Adhesion of stored RBC to the vascular endothelium has been shown to lead to impaired perfusion in the microcirculation and contribute to organ failure and poor outcome. The role of either or both of the EC and RBC glycocalyx in this process is unknown and was studied in an in vitro model. Human umbilical vein endothelial cells were perfused in a microfluidic device with RBC solutions from fresh, less than 14-day or longer than 21-day storage. In some experiments, the HS microenvironment was simulated by hypoxia-reoxygenation (H/R) and epinephrine (Epi) in the perfusion experiments. Measurements obtained included endothelial cell (EC) and RBC glycocalyx and RBC adherence to human umbilical vein endothelial cell monolayers at variable shear rates. Endothelial cell glycocalyx and RBC glycocalyx dimensions were reduced by H/R and Epi and storage duration respectively. Red blood cell adherence to the endothelium was increased by H/R + Epi treatment and duration of RBC storage. Our data may help explain some of the remaining discrepancies regarding the impact of RBC storage duration on outcomes in the trauma population. Consideration of the integrity of the EC and RBC glycocalyx may guide future transfusion strategies in the trauma population. The microfluidic device system platform may offer a high throughput modality to study emerging therapies to mitigate adverse consequence of RBC storage duration on the perfused endothelium in the trauma setting.

Use of potassium adsorption filter for the removal of ammonia and potassium from red blood cell solution for neonates.

Ammonia in the plasma usually does not pass through the blood-brain barrier (BBB). However, it can affect the brain as a neurotoxin in neonates with anemia of prematurity. Excess intake of ammonia should therefore be restricted in conditions involving BBB breakdown, such as in premature neonates. A potassium adsorption filter (PAF) can remove not only potassium, but also ammonia from red blood cell (RBC) solution. PAF for neonates (PAF-n) has been recently introduced using small satellite packs. We evaluated the effects of PAF-n on the removal of ammonia and potassium from RBC solution in small satellite packs. RBC solutions were obtained from the Japanese Red Cross Society. Two units of RBC solution (280 mL) were divided into four satellite packs (70 mL/pack). The RBC solution was passed through PAF-n (Kawasumi Laboratories Inc.) that was primed with saline (100 mL) before use. The concentrations of ammonia and potassium were measured in the solution before and after filtration (four samples of 10 mL each of filtered RBC solution) by Biomedical Laboratories. Approximately 47 to 82 and 84% to 93% of ammonia and potassium were removed from the RBC solution, respectively, without dilution with saline. PAF-n can remove ammonia and potassium from RBC solution in small satellite packs. PAF-n could therefore improve the clinical prognosis of neonates with poorly developed BBB by limiting the delivery of excess ammonia found in the RBC solution.

Measuring osmosis and hemolysis of red blood cells.

Since the discovery of the composition and structure of the mammalian cell membrane, biologists have had a clearer understanding of how substances enter and exit the cell's interior. The selectively permeable nature of the cell membrane allows the movement of some solutes and prevents the movement of others. This has important consequences for cell volume and the integrity of the cell and, as a result, is of utmost clinical importance, for example in the administration of isotonic intravenous infusions. The concepts of osmolarity and tonicity are often confused by students as impermeant isosmotic solutes such as NaCl are also isotonic; however, isosmotic solutes such as urea are actually hypotonic due to the permeant nature of the membrane. By placing red blood cells in solutions of differing osmolarities and tonicities, this experiment demonstrates the effects of osmosis and the resultant changes in cell volume. Using hemoglobin standard solutions, where known concentrations of hemoglobin are produced, the proportion of hemolysis and the effect of this on resultant hematocrit can be estimated. No change in cell volume occurs in isotonic NaCl, and, by placing blood cells in hypotonic NaCl, incomplete hemolysis occurs. By changing the bathing solution to either distilled water or isosmotic urea, complete hemolysis occurs due to their hypotonic effects. With the use of animal blood in this practical, students gain useful experience in handling tissue fluids and calculating dilutions and can appreciate the science behind clinical scenarios.

Relative Sensitivities of Japanese Quail to Foreign Red Blood Cell Challenges for Immunotoxicity Testing

Given evidence that the immune system is sensitive to environmental contaminants, evaluating immunocompetence in toxicology studies is increasingly important. By incorporating a test of humoral response into controlled reproductive and developmental studies, more comprehensive results can be gathered to assess the potential for disease. The foreign red blood cells (RBC) challenge is a minimally invasive method for evaluating humoral responses to a foreign antigen. Typically, antibody response is assessed following injections of sheep erythrocytes; however, Japanese quail (Coturnix japonica) are only minimally sensitive to sheep RBC. In the present study, adult Japanese quail were treated with 5% solutions of RBC from goose, goat, cow, donkey, or pig in primary and secondary challenges. After each treatment, plasma samples were taken and antibody responses were measured for total immunoglobulins (Ig), IgG, and IgM. Overall, goose RBC generated the poorest responses in both primary and secondary challenges, while the strongest antibody responses were to pig and donkey RBC. Therefore, pig RBC appear to be a superior antigen for testing humoral response in Japanese quail.

Sensitive and Simple Methodologies for Measuring of Red Blood Cell (RBC) Electrical Properties and Cell Aggregation.

The red blood cell membrane contains proteins and glycoproteins embedded in a fluid lipid bilayer that confers viscoelastic behaviour. Sialylated glycoproteins of the RBC membrane are responsible for a negatively charged surface, which creates a repulsive electric zeta potential (ζ) between the cells.The compact layer of charge or Stern consists of ions rigidly bonded to the cell and the double layer includes ions diffusely distributed around the cell. Measurement of RBC layer of charge and zeta potential can be use as a marker of RBC membrane injury. Study of membrane electrical properties and forces and bindings between cells, can provide a better comprehension of membrane injury presented in RBC unit storage, after RBC unit irradiation, cell senescence process and in addition, also to improve the sensitivity and specificity of the hemoagglutination reaction. The aim of this study was to measure the RBC double layer thickness of the charge (DLC) around the cell, zeta potential (ζ) and cell aggregation force, using optical tweezers.To measure DLC (d) and zeta potential (ζ) a special chamber was built consisting of 2 platinum electrodes separated by a channel. An external electrical field (E) was applied with a voltage power supply connected to the electrodes. The optical tweezers consisted of Nd:YAG laser beam focused through the 100x objective of a microscope equipped with a minicam, which registered the trapped cell image in real time. These images were recorded in VHS and captured in a computer where they could be analyzed using Image Pro Plus software. Optical tweezers utilizes the light momentum transfer to trap the silica beads with pico Newtons forces (10−12). Ten RBC units from healthy blood donors were collected in CPDA-1. Samples were diluted (1:1000μl) in AB plasma. Silica beads were added to a RBC solution (10μl). DLC thickness (d) was obtained measuring, in the special chamber, the force on the silica bead attached to a single RBC in response to an applied voltage (50–100V). Zeta potential (ζ) was obtained by measuring the RBC terminal velocity (Tv) after being released from the optical trap at a constant voltage applied supply (100V) Tv=(εE/η) ζ; ε= medium dielectric constant; η= viscosity). The DLC was calculated by the equation Fop= (Aεζ/d)E (Fop= optical force; A= RBC area). DLC median size (d) was 0,85 μm (min 0.4/max 1.3) and zeta potential was ζ=−12,5 mV (min- 9.3/max −15). These results were in accordance with the data in literature, where other methods to measure the same property were used.For measuring RBC aggregation, a silica bead attached to RBCs was trapped and the force needed to slide one RBC over the other, as a function of the velocities (2,5,7,12,18,32X10−7m/s) was determined. RBC O Rh(D) positive were diluted (1:1000μl) in plasma AB (control group) and in agglutination potentiator solutions, used in immunohematological tests (dextran 0.15%, low ionic strength solution-LISS, bromelain and papain enzymes). Aggregation was also analyzed using anti-D antibodies serum (5μl). The median aggregation force obtained of 20 RBC control samples was 1x10−3 (0.1–2.5) poise.cm. Samples analyzed with dextran 0.15%, LISS and bromelain lacking antibodies showed no agglutination. RBCs treated with papain enzyme presented high adherence preventing the capability of separating cells. The 20 samples analyzed with anti-D serum showed median aggregation force 2x10−3 (1.0–4.0) poise.cm. These results obtained from RBC with antibodies were statistically higher (p<0.001) when compared to control. The method revealed to be sensitive and results were in accord with serological tests used in transfusionroutine, where false positive results could be frequently found in RBC treated with papain, however rarely obtained with dextran and bromelain. In conclusion, the methodologies proposed are simple and may provide specific information on RBC interaction in addition to cell aggregation. Individual cell analyze methods are always more sensitive to small differences than those based on average values. This is the first time that the size of the double layer was measured using optical tweezers and the ability of the optical trap also allowed us to obtain the zeta potential and DLC thickness for the same RBC.

Practical application of NMR spectra of 129Xe dissolved in red blood cell

The 129Xe NMR spectra in red blood cell (RBC) were investigated in vitro focusing upon temperature dependence and the magnetization transfer (MT) experiment. 129Xe signals were observed at 192 and 201 ppm in the RBC solution, which were assigned to 129Xe dissolved in saline and RBC phases, respectively. A presence of the third peak was evidenced by the MT experiment, in which a pre-saturation around 210–220 ppm caused a decrease in the intensity of the 192-ppm peak. To examine the applicability of the 129Xe NMR spectra index for the detection of physiological conditions, the temperature dependence of the chemical shift was studied precisely and accurately in the RBC solution. The multinuclear internal reference method was adopted to express the chemical shift on an absolute frequency scale. The temperature dependence was proved to be non-linear and approximated by a convex curve for the 192 ppm peak, whereas it was linear for the 201-ppm peak. The temperature coefficient amounted to as much as 0.2 ppm/°C (5 °C/ppm) for the 129Xe signal in RBC. A quite similar coefficient was observed for the separation of 192 and 201 ppm peaks, which was measured simply without the adoption of an absolute frequency scale and will be suited for practical application.

Behaviour of Erythrocyte Suspensions in Flow Through a Branched Tube. A Study on the Deposition Hypothesis of Atherogenesis

Introduction Predilection sites of atherosclerosis and thrombotic events are the inner walls of curved and the outer walls of branched arteries. At these sites similiar hydrodynamic situations exist: elevated hydrostatic pressure, stagnating flow tending to separate, secundary flow or turbulence. Such flow conditions may be involved in atherogenesis by increasing the transport of atherogenetic substances into the wall or depositing blood elements onto the endothelium. Both concepts are still hypothetically. The aim of our experiments was only to answer one of the unanswered questions: can flow conditions occurring in branched tubes cause deposits of suspended blood cells on (impermeable) tube walls, and what are the hydrodynamic mechanisms involved? The presented experiments deal with the motion and distribution of red blood cells (RBC) studied in stationary branched flow. Corresponding “micro-hydrodynamic” local flow conditions have been studied in parallel experiments. Materials and Methods Glass models: 1. straight glass tubes (ID 3.0 mm, wall thickness 0.2 mm) with a side branch (ID 1.5 mm; branch angle 90°); 2. tubes with rectangular section area (2.0 × 1.5 mm) side branch (ID 1.5 mm; branch angle 90°) used for sedimentation experiments; 3. T-tube to produce a stagnation point flow: the inflow tube (ID 3.45 mm) leads the fluid vertically on to the window of a flow chamber (170 × 10 × 5 mm); distance between inflow tube and window was 2.7 mm. Suspensions: A. Once saline-washed human RBC in isosmotic saline solutions of Dextran (Rheomacrodex®, 6 g%; MW 40,000). B. Washed human RBC in saline solutions of Polyvinylpyrrholidon (1 g%, MW 360,000) added to make the RBC “sticky”. C. RBC in solutions of 1.5 g% Polyvinylpyrrholidon (360,000). Suspensions A and B contained 5 vol% of homologous plasma. Viscosity of suspensions A, B and C: η = 2.6 cp, 2.6 cp and 5.4 cp resp.; volume fraction RBC c ≈ 0.01. Flow apparatus: A Marione bottle (reservoir for the suspension) kept the hydostatic pressure constant. From here the suspension flowed through a stabilisation tube into the branched tube. The branches were connected to separated cylinders to measure the volume flow per time through the main (Q 1) and the side (Q 1) branch. The main tube was mounted vertically in all except in sedimentation experiments. Particle behaviour within the glass models 1 and 2 and in the stagnation point flow was studied by dark field microscopy. By means of a laser ultra-microscope (Kratzer and Kinder, 1975), velocities of RBC in the stagnation point flow were measured. Results and discussion Due to “radial migration” in undisturbed straight tube flow (Q 1 = 0) a RBC free layer adjacent to the wall prevents RBC from contact with the wall (Fig. 2, 3 a). By any flow Q 1 > 0 the thickness of this layer is altered in a definite region opposite the orifice of the side branch. Mere widening is observed with small Q 2 (Fig. 3 b). For Q 2 exceeding a critical value the layer vanishes in the region of reattachment. At the same site “sticky” RBC are deposited, Fig. 4, being torn off with larger Q 2. The flow at the sites of RBC concentrated near wall or deposited can be described as stagnation point flow (Fig. 1), which is characterized by ventrical (V 1) and wall-parallel (V //) velocity components. The V I and V // necessary to allow deposis can be measured in a rotary symmetric stagnation point flow. Under such flow conditions the V IE and V //E of RBC necessary to deposit are: V IE ⩾ 0.45 mm/s 0.3 mm distant from the wall; V //E < 0.05–0.13 mm/s (wall shear stress τ W < 0.27–0.7 dyn/cm 2). Similiar values have been measured in the branched tube (Fig. 5). Figure 7 shows pathways and velocities of RBC in the stagnation point flow, forming stripes of deposits on the flat cover glass of the perfused T-tube. Deposits formed by sedimented RBC in a branched tube flow are shown and explained in figure 6. The results indicate that the formation of RBC deposits depend on the existence of velocity components V I normal to and against the wall of the perfused tube. Minimum V I are necessary to bring the RBC to the wall (s. a. Forstrom et al., 1974). If moreover the parallel velocity component V // does not exceed a certain value, deposits can be formed (s. a. Petschek and Weiss, 1970). Deposition sites in branched glass tubes correspond to predilection sites of atherosclerosis in branched arteries (e.g. Wolkoff, 1929).