Preservation Of Red Blood Cells Research Articles

Profiles of differential expression of miRNAs in the late stage of red blood cell preservation and their potential roles

ObjectiveTo detect the differentially expressed regulatory miRNAs in the late stage of red blood cell (RBC) preservation and predict their roles. MethodsSuspended RBCs with different storage periods of 35 day, 42 day, and 50 day were collected for routine blood tests, RNA extraction, and preparation of small RNA sequencing libraries. The constructed libraries were sequenced and the biological functions of differential miRNAs in RBCs in the late storage were analyzed by bioinformatics. ResultsRoutine indicators of RBCs in the late stage were not significantly affected by preservation time. The Pearson correlation analysis performing on RBC miRNAs with different storage days revealed that RBC miRNAs changed with the increase of storage days. RBC miRNAs from day 35 (D35), day 42 (D42) and day 50 (D50) showed significant differences (P<0.05). Compared RBC miRNAs from D42 with these from D35, there were 690 up-regulated miRNAs and 82 down-regulated miRNAs; compared RBC miRNAs from D50 with these from D35, there were 638 up-regulated miRNAs and 123 down-regulated miRNAs; compared RBC miRNAs from D42 with these from D50, there were 271 up-regulated miRNAs and 515 down-regulated miRNAs. GO enrichment analysis of target genes of differential miRNAs were mainly involved in cell metabolism, biosynthesis, protein modification, gene expression and transcriptional regulation of biological processes. KEGG pathway enrichment analysis of miRNA target genes showed that differential miRNA target genes were closely related to pathways in cancer. ConclusionMiRNAs were differentially expressed in the late stage of RBC preservation, and may be involved in various biological processes, especially cancer.

In vitro Evaluation of DINCH-Plasticized Blood Bags for Red Blood Cell Storage with CPDA-1 Anticoagulant

Plain Language SummaryA plasticizer is a chemical substance added to plastic to increase its flexibility. DEHP is a plasticizer that has been widely used in many products including plastic tubing and bags of medical devices. However, concerns about DEHP-related toxicity have been debated for many years. DEHP has been replaced with other plasticizers in many products, but it is still being used in blood bags due to its protective effect on RBC preservation. DINCH is an alternative plasticizer with a low toxicology profile. This study investigated the quality of RBC concentrates stored in blood bags using DINCH. Twenty sets of five RBC concentrates were produced using two DINCH bags and three DEHP bags with CPDA-1 anticoagulant, and the storage quality was assessed weekly for 35 days. On day 35, the median hemolysis levels in the DINCH bags (0.297–0.342%) were slightly increased than the DEHP bags (0.204–0.240%). However, all DINCH bags showed hemolysis lower than the regulatory limit of 0.8%. DINCH bags exhibited better gas permeability than DEHP bags. Compared to DEHP, DINCH exhibited considerably lower levels of plasticizer leaching into blood bags. Most of the other metabolic parameters were comparable in both bags. The quality of nonleukocyte-reduced RBC concentrates stored for 35 days in DINCH-plasticized blood bags with CDPA-1 is generally comparable to those in DEHP bags. Hence, DINCH can be a viable alternative to DEHP in blood bags for RBC storage, even without the use of next-generation additive solutions to improve RBC preservation quality.

Long-term preservation of red blood cells through deep supercooling

Long-term preservation of red blood cells through deep supercooling

Gestation outcomes in various options of help for pregnant women with Rh-immunization

BACKGROUND: To date, several options for helping pregnant women with rhesus immunization are known: (a) active tactics in carrying out methods of efferent therapy in the form of basic operations (plasmapheresis, hemosorption) in combination with adjuvant methods (immunoglobulin, blood photomodification with ultraviolet, laser beams, ozone therapy) to pregnant women; (b) wait-and-see active tactics with observation of the pregnant woman, followed by intrauterine intravascular transfusions of washed donor red blood cells; (c) mixed active tactics with a sequential combination, alternation of these methods. In Russia, only option 2 with fetal transfusions of washed donor red blood cells is accepted as the basis and paid for. The objective of the study is to conduct a comparative analysis of pregnancy outcomes in women with rhesus immunization using different management options.&#x0D; MATERIALS AND METHODS: A total of 392 women were followed up at seven different institutions in Russia and at the Donetsk Center for Maternal and Child Health (DNR), of whom 345 pregnant women (Group 1) received efferent therapy, 33 women (Group 2) had fetuses intrauterine bypass surgery, and 14 pregnant women (Group 3) had mixed efferent therapy and fetal PEEP bypass surgery.&#x0D; RESULTS: The analysis showed that the most favorable results for the main clinical indicators (premature, operative delivery, fetal hypoxia at birth, etc.) were in Group 1 and 3 women, in which the perinatal mortality was 14.5/1000 and 0/1000, respectively, which was significantly lower than in Group 2 (176.5/1000). It was also found that in Groups 2 and 3 women, the mean intervals between repeated transfusions of washed donor red blood cells were 8.8 0.2 and 21.4 3.8 days (p 0.01), which may be explained by the detoxifying effect of efferent therapy methods, preservation of fetal red blood cells and transfused donor red blood cells to the fetus with prolonged gestation and obtaining healthier and more viable progeny.&#x0D; CONCLUSIONS: 1. Severe Rh conflict is a manifestation of a syndrome of systemic effects of aggressive metabolites of specific and nonspecific nature. 2. The etiopathogenetic measure in the prevention and treatment of HDF/HDN in rhesus conflict is efferent therapy methods for the mother, and transfusion of washed donor rhesus-negative red blood cells to the fetus is effective, but a temporary, palliative measure, as is the case in multiple organ failure. 3. In the treatment protocols, efferent therapy methods must be present to prevent fetal red cell destruction and, equally importantly, to prevent destruction of Rh-negative donor red cells transfused to the fetus.

The development and application of a novel reagent for fixing red blood cells with glutaraldehyde and paraformaldehyde

ABSTRACTObjectiveThe currently employed red blood cell reagents have a short shelf life. Some hospitals with a small number of specimens will be unable to utilize them within the validity period, resulting in a substantial increase in the purchase price. Therefore, the method of developing long-term red blood cell reagents is a problem worthy of further study.MethodsIn this experiment, the type and concentration of the red blood cell reagent treatment solution were evaluated based on the red blood cell antigen concentration 24 h after treatment. In addition, the qualified glutaraldehyde/paraformaldehyde reagent was stored for six months, and five red blood cell indices were measured every month. At the same time, the detection indices of treated red blood cell reagents and untreated red blood cell reagents were compared.ResultsIt was discovered that treated red blood cells containing 0.005% GA and 0.05% PFA were more suitable for the preservation of red blood cells than other treated concentrations, and the preservation time could reach six months. The test tube method (n = 24) and microcolumn gel card (n = 35) were used to determine the accuracy of the treated blood cells containing 0.005% glutaraldehyde +0.05% paraformaldehyde, with an accuracy of 100%.ConclusionThis experiment resulted in the development of a novel reagent for treating red blood cells with glutaraldehyde/paraformaldehyde fixed solution that can effectively prolong its storage time by two to three times that of red blood cell reagents currently on the market.

Preservation of red blood cell antigenicity in a new storage solution in vitro

Introduction Red blood cell (RBC) storage solution is used for suspending and preserving RBCs for later use in in vitro immunohematology testing. Proper RBC preservation is crucial for obtaining accurate results in RBC phenotyping and pretransfusion antibody screening tests. Haemolysis or RBC antigen degradation during storage can result in inaccurate RBC phenotyping, thereby decreasing the sensitivity of pretransfusion antibody screening and identification assays. The conventional RBC storage solutions usually contain adenosine, adenine, and antibiotics. We designed an RBC storage solution and determined whether it could preserve RBC integrity for 70 days. Materials and Methods The new storage solution has a different formula from that of the conventional solution—in particular, it is strengthened with polyethylene glycol (PEG). The extent of haemolysis and hemagglutination reactivity of the RBC antigen systems, Rh, Duffy, Kidd, Lewis, MNS, P1, and the rare antigen Mia (which has a low prevalence antigen in most parts of the world but a higher prevalence in Taiwan), in the new RBC storage solution was compared with that of the conventionally preserved RBC storage solution. Results The RBCs preserved in the new solution for 70 days retained a similar haemolysis grade as those preserved in the control solution for 28 days. Although both solutions largely preserved RBC antigenicity, the decline in RBC hemagglutination scores in new solution often occurred later than that in the control solution in most antigen phenotyping assays, especially labile antigens such as D, P1, and M. Conclusion The new solution reduces haemolysis more effectively and preserves antigenicity throughout the 70-day storage period. Moreover, Mia antigen is more stable in the experimental group.

Advantages of pyruvate-based fluids in preclinical shock resuscitation-A narrative review.

This review focuses on the innate beneficial effects of sodium pyruvate-based fluids, including pyruvate in intravenous solutions, oral rehydration solutions, and peritoneal dialysis solutions, on shock resuscitation with various animal models relative to current commercial fluids over the last two decades. Due to its superior pharmacological properties, pyruvate effectively sustains cytosolic glycolytic pathways and mitochondrial oxidative phosphorylation by restoration of redox potentials and reactivation of pyruvate dehydrogenase in hypoxia, even anoxia, and diabetes, reversing the Warburg effect and diabetic glucometabolic aberration. Pyruvate has been demonstrated to protect against multiorgan dysfunction and metabolic disturbance in numerous preclinical studies with various pathogenic injuries. The unique features of pyruvate potential clinical benefits encompass to efficiently correct lethal lactic acidosis via metabolically rapid consumption of intracellular [H+] and robustly protect multiorgan metabolism and function, particularly visceral organs in addition to the heart and brain, significantly prolonging survival in various animal models. Pyruvate protection of red blood cell function and preservation of the partial pressure of arterial oxygen should be highly concerned in further studies. Pyruvate is much advantageous over existing anions such as acetate, bicarbonate, chloride, and lactate in commercial fluids. Pyruvate-based fluids act as a therapeutic agent without causing iatrogenic resuscitation injury in addition to being a volume expander, indicating a potential novel generation of resuscitation fluids, including crystalloids and colloids. Pyruvate-based fluids have an enormous potential appeal for clinicians who face the ongoing fluid debate to readily select as the first resuscitation fluid. Clinical trials with pyruvate-based fluids in shock resuscitation are urgently warranted.

Changes of Lipid Peroxidation after Administration of Nanomodified Resuspending Solution into Donor Red Blood Cells

The problem of increasing the functional activity preservation of red blood cells is priority for the Blood Service and clinical transfusiology. Taking into account the previously discovered unique properties of magnetite nanoparticles, namely: nonspecific modulation of metabolic processes, strengthening of adaptive mechanisms of cellular organelles, acceleration of reparative processes at the level of membranes and macromolecules, for the first time an attempt was made to use the upgraded saline solution of sodium chloride (nanotechnologically upgraded solution - NUS) as a resuspending of erythrocyte-containing components (ECC). Objective: to improve the preservation of red blood cells containing donor blood components using NUS for resuspending. To achieve this goal, the first stage of the study examined the effect of a new resuspending solution (NUS) on lipoperoxidation, catalase activity, and red blood cell peroxidation resistance in donor blood components containing red blood cells. Object of Research: red blood cells from preservation blood of donors, which are intended for component transfusion therapy. As a result of the study, the advantages of using NUS were established, namely: a significant decrease in the levels of primary and secondary lipid peroxidation products in the first 15 days of observation, an increase in catalase activity and resistance of red blood cells to peroxidation in the first stages of the study, inhibition of the shear reaction with neutral lipid peroxidation in the late stages of observation.

NonFreezable Preservation of Human Red Blood Cells at -8 °C.

Red blood cell (RBC) preservation is very important in human health. The RBCs are usually preserved at 4 ± 2 °C without freezing or at a very low temperature (-80 °C or liquid nitrogen) with deep freezing. Herein, non freezable preservation of RBCs at a subzero temperature is reported to prolong the preservation time compared with that at 4 ± 2 °C. By adding glycerol and poly(ethylene glycol) (PEG) (average number molecular weight 400, PEG-400) into the preservation solution, the freezing point is decreased and the hemolysis is kept low. The cell metabolism of stored RBCs at -8 °C is reduced, and the shelf life of RBCs extends up to at least 70 days. At the end of preservation, the pH decreases a little bit to demonstrate the low metabolic rate of RBCs stored at subzero temperatures. After quick washing, the RBC survival rate is ca. 95%. The adenosine triphosphate, 2,3-diphosphoglycerate, and cell deformation ability of the washed RBCs are maintained at a high level, while the malondialdehyde is relatively low, which verifies the high quality of RBCs stored at this condition.

"Study of Effects a New Resuspending Solution Which Was Nanotechnologically Upgraded on Lipoperoxidation, Catalase Activity and Red Blood Cell Peroxidation Resistance in Donor Blood Components"

The problem of increasing the functional activity preservation of red blood cells is priority for the Blood Service and clinical transfusiology. Taking into account the previously discovered unique properties of magnetite nanoparticles, namely: non-specific modulation of metabolic processes, strengthening of adaptive mechanisms of cellular organelles, acceleration of reparative processes at the level of membranes and macromolecules, for the first time an attempt was made to use the upgraded saline solution of sodium chloride (nanotechnologically upgraded solution - NUS) as a resuspending of erythrocyte-containing components (ECC).

The role of corticosterone in the regulation of the cellular composition of chicken blood during the stress reaction

The influence of hen layer density on the variability of the number of red blood cells, heterophiles and lymphocytes in the blood, the secretory activity of adrenal glands, estimated by the level of corti-costerone and cortisol, as well as the presence of interrelations between hormones and blood cells by calculating complex indices, were studied. Chickens, as the research object, were kept in cages, under conditions of standard layer density and increased by 1.5 and 2.0 times. We found that chickens adapt to an increase in layer density by one and a half times, pro-vided that egg production decreases to 33.33%; two times exceed of the regulatory requirements for laying does not correspond to the adaptive abilities of birds. Depending on the level of layer density excess (stress factor) in chicken blood, the concentration of corticosterone and cortisol increases, determining a decrease in the number of lymphocytes and an in-crease in heterophiles against the background of the preservation of red blood cells, reflecting the “energy price” of adaptation. Corticosterone af-fects the relationship of red blood cells with lymphocytes and heterophiles, determining the variability of the values of the indices reflecting the ratio of red blood cells and lymphocytes (ISEL), red blood cells and hetero-philes (ISEG), red blood cells, lymphocytes and corticosterone (ISELC), red blood cells, heterophiles and corticosterone (ISEGC) and the integral index of red blood cells-heterophiles-lymphocytes and corticosterone (IIEGLC).

Hydrogel Microfiber Encapsulation Enhances Cryopreservation of Human Red Blood Cells with Low Concentrations of Glycerol.

Cryopreservation of red blood cells (RBCs) has been studied as a typical example of cryobiology methodology. To date, a mature and long-term cryopreservation process for RBCs has been developed, which has the weakness of complicated procedures due to high concentrations of glycerol (Gly). Therefore, it is still a research focus to find a new method for cryopreservation of RBCs to reduce the concentrations of cryoprotectants (CPAs). In this study, alginate hydrogels, which have been widely used in preservation research, were selected because of their advantages such as lower cost, cytocompatibility, and crosslinking occurring under mild conditions. With a variety of CPA solutions, the RBC recovery with the cryopreservation of RBCs and RBC-laden hydrogel microfibers was compared in different situations. It was found that hydrogel microfiber encapsulation enhances cryopreservation of human RBCs with low concentrations of CPAs. And Gly can be removed by washing directly with a 0.9% sodium chloride solution. This study validated the feasibility of cryopreservation of RBC-laden hydrogel microfibers. It may provide a new and evolving direction for reducing the concentrations of CPAs used in the preservation of RBCs.

Application of Nanotechnology for the Preservation of Red Blood Cells

Application of Nanotechnology for the Preservation of Red Blood Cells

Long-term deep-supercooling of large-volume water and red cell suspensions via surface sealing with immiscible liquids

Supercooling of aqueous solutions is a fundamentally and practically important physical phenomenon with numerous applications in biopreservation and beyond. Under normal conditions, heterogeneous nucleation mechanisms critically prohibit the simultaneous long-term (> 1 week), large volume (> 1 ml), and low temperatures (< −10 °C) supercooling of aqueous solutions. Here, we report on the use of surface sealing of water by an oil phase to significantly diminish the primary heterogeneous nucleation at the water/air interface. We achieve deep supercooling (down to −20 °C) of large volumes of water (up to 100 ml) for long periods (up to 100 days) simultaneously via this approach. Since oils are mixtures of various hydrocarbons we also report on the use of pure alkanes and primary alcohols of various lengths to achieve the same. Furthermore, we demonstrate the utility of deep supercooling via preliminary studies on extended (100 days) preservation of human red blood cells.

The Chemistry of Lyophilized Blood Products.

With the development of new biologics and bioconjugates, storage and preservation have become more critical than ever before. Lyophilization is a method of cell and protein preservation by removing a solvent such as water from a substance followed by freezing. This technique has been used in the past and still holds promise for overcoming logistic challenges in safety net hospitals with limited blood banking resources, austere environments such as combat, and mass casualty situations where existing resources may be outstripped. This method allows for long-term storage and transport but requires the bioconjugation of preservatives to prevent cell destabilization. Trehalose is utilized as a bioconjugate in platelet and red blood cell preservation to maintain protein thermodynamics and stabilizing protein formulations in liquid and freeze-dried states. Biomimetic approaches have been explored as alternatives to cryo- and lyopreservation of blood components. Intravascular hemostats such as PLGA nanoparticles functionalized with PEG motifs, topical hemostats utilizing fibrinogen or chitosan, and liposomal encapsulated hemoglobin with surface modifications are effectively stored long-term through bioconjugation. In thinking about the best methods for storage and transport, we are focusing this topical review on blood products that have the longest track record of preservation and looking at how these methods can be applied to synthetic systems.

Pilot study on novel blood containers with alternative plasticizers for red cell concentrate storage.

Di (2-ethylhexyl) phthalate (DEHP), a typical plasticizer used for polyvinyl chloride (PVC) blood containers, is eluted from the blood containers and exerts protective effects on red blood cells. However, a concern for detrimental effects of DEHP on human health has led to the development of potential DEHP substitutes. Here, we compared the red blood cell preservation ability of two types of non-DEHP blood containers with safe alternative plasticizers to that of DEHP blood containers. Red cell concentrates in mannitol-adenine-phosphate solution (MAP/RCC) were stored for 6 weeks in PVC blood bags containing DEHP, di-isononyl-cyclohexane-1,2-dicarboxylate (DINCH) and di (2-ethylhexyl) 4-cyclohexene-1,2-dicarboxylate (DOTH), or 4-cyclohexene-1,2-dicarboxylic acid dinonyl ester (DL9TH) and DOTH. There was no significant difference in the total amount of plasticizer eluted into MAP/RCC (till 3 weeks from the beginning of the experiment), hemolysis of MAP/RCC, and osmotic fragility of MAP/RCC between the non-DEHP blood containers and DEHP blood containers. Hematological and blood chemical indices of MAP/RCC in all containers were nearly the same. Thus, DOTH/DINCH and DOTH/DL9TH blood containers demonstrate the same quality of MAP/RCC storing as the DEHP blood containers. Since DOTH, DINCH, and DL9TH were reported to be safe, DOTH/DINCH and DOTH/DL9TH blood containers are promising candidate substitutes for DEHP blood containers.

The immune-stimulation capacity of liposome-treated red blood cells

Our in vivo studies on a rat model established safety of transfusing liposome-treated red blood cells (RBCs) but identified the potential for immune modulation as related to transfusion efficacy of liposome-treated RBCs. The aim of this study was at assessing the impact of liposome-induced membrane changes on the immune profile of liposome-treated RBCs by (a) evaluating their interaction with endothelial cells and monocytes; and (b) the resulting immune response derived from this interaction, in the form of cytokine release, adhesion molecules expression and phagocytosis. Unilamellar liposomes were synthesized to contain unsaturated phospholipids (1,2-dioleoyl-sn-glycero-3-phosphocholine [DOPC]:CHOL, 7:3 mol%). The human RBCs immune profile was assessed by incubating control and DOPC-treated RBCs with human umbilical vein endothelial cells (HUVECs) and monocytes. Cytokine release measured by Luminex technology, vascular cell adhesion molecule (VCAM)-1 and E-selectin on HUVECs measured by flow cytometry, and the erythrophagocytic activity of monocytes by monocyte monolayer assay (MMA) were determined. Fibroblast growth factor [FGF]-2 was the only cytokine released by HUVECs that remained increased after incubation with DOPC-treated RBCs compared to control throughout storage. The expression of both VCAM-1 (15.3 ± 5.6% versus 6.3 ± 0.9%, p = 0.008) and E-selectin (18.0 ± 6.3% versus 6.6 ± 0.7%, p = 0.004) by HUVECs were significantly increased after incubation with DOPC-treated RBCs at day 2 of storage. The MMA resulted in phagocytic indexes of zero for both control and DOPC-treated RBCs at day 2 and 42 of storage. The liposome treatment did not result in significant changes to the immune profile of stored DOPC-treated RBCs. These findings combined with previous in vivo results, make liposome treatment a potential candidate for application in RBC preservation and open the possibility for clinical use with other cell types.

Assessment of physical damage of cryopreserved RBCs during thawing by impedance spectroscopy

Background: Preservation of red blood cells (RBCs) is an important task to ensure a long-term, readily available, safe blood supply for transfusion during emergency. Effective preservation procedures are required at various steps for the long-term storage of an RBC product including testing, inventory, quality control, and product distribution. Biopreservation is the process of maintaining the integrity and functionality of cells outside the native environment for long storage times. Hypothermic storage, cryopreservation, and lyophilization are the various methods of biopreservation. Objective: To monitor the physical integrity of erythrocytes after cryopreservation with the help of impedance spectroscopy. Materials and Methods: Cryoprotectants were added to protect the erythrocytes from cryoinjuries. In addition, the erythrocytes viability was monitored through hemolysis test. Afterward, the exact numbers of live or lysed cells were observed by the effect of thawing rate on the cells through impedance spectroscopy, as we know that there is a relationship between impedance, frequency, and temperature. Result: As the frequency increases, the impedance decreases, which indicates that the RBCs are lysed. The impedance starts decreasing at −200 °C and drastically decreased at −140 °C. Conclusion: This study indicates the real viability of the RBCs through impedance measurement by the impedance spectroscopy.

Construction of d-α-tocopheryl polyethylene glycol succinate/PEO core-shell nanofibers on a blood-contacting surface to reduce the hemolysis of preserved erythrocytes.

The hemolysis of erythrocytes is a big obstacle to the development of new non-plasticizer polymer containers for erythrocyte preservation. To construct a long-term anti-hemolytic surface of a plasticizer-free polymer, we coaxially electrospin core-shell structured d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS)/poly(ethylene oxide) nanofibers on the surface of a styrene-b-(ethylene-co-butylene)-b-styrene (SEBS) elastomer that is covered with grafted poly(ethylene glycol) (PEG) chains. Our strategy is based on the fact that the grafted layers of PEG reduce mechanical damage to red blood cells (RBCs) while the TPGS released from the nanofibers on a blood-contacting surface can act as an antioxidant to protect RBCs from oxidative damage. We demonstrate that TPGS/PEO core-shell structured nanofibers have been well prepared on the surface of PEG modified SEBS; the controlled release of TPGS in distilled water is obtained and the release can last for almost 4 days at 4 °C; during RBC preservation, TPGS acts as the antioxidant to decrease the membrane oxidation and hemolysis of RBCs. Our work paves a new way for the development of non-plasticizer polymers for RBC preservation, which may be helpful for the fabrication of long-term anti-hemolytic biomaterials in vivo.

Preservation of Red Blood Cells in the American Alligator (Alligator mississippiensis) over Time and in Two Different Anticoagulants

Abstract Lesions associated with storage of red blood cells (RBC) in different anticoagulants have been well described in mammals, but not in reptiles to date. The objectives of this study were to document biochemical changes of American alligator (Alligator mississippiensis) packed RBC stored in citrate–phosphate–dextrose–adenine (CPDA-1) and acid–citrate–dextrose (ACD) anticoagulants over the course of 35 days and to describe qualitative and quantitative morphologic changes over 295 days. Blood samples were collected via the occipital sinus from 6 captive-born American alligators of mixed sex (1 male, 5 females). Spun packed cell volume (PCV), total solids, and blood lead levels were analyzed with the use of heparinized blood on Day 0. Samples in CPDA-1 and ACD were stored at 4°C (39.2°F) and the following parameters serially evaluated at varying time points until Day 35: blood culture, blood gas analysis, PCV, methemoglobin, osmotic fragility, and morphologic evaluation by light and transmission electr...