Research | Volume 39, Article 280, 30 Aug 2021 | 10.11604/pamj.2021.39.280.25695

Evaluation of cellular changes in blood stored for transfusion at Bungoma County Referral Hospital, Kenya

Phidelis Maruti Marabi, Stanslaus Musyoki, Angela Amayo

Corresponding author: Phidelis Maruti Marabi, Bungoma County Referral Hospital, Bungoma, Kenya

Received: 22 Aug 2020 - Accepted: 12 Aug 2021 - Published: 30 Aug 2021

Domain: Haematology

Keywords: Blood transfusion, cellular changes, storage, Kenya

©Phidelis Maruti Marabi et al. Pan African Medical Journal (ISSN: 1937-8688). This is an Open Access article distributed under the terms of the Creative Commons Attribution International 4.0 License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Cite this article: Phidelis Maruti Marabi et al. Evaluation of cellular changes in blood stored for transfusion at Bungoma County Referral Hospital, Kenya. Pan African Medical Journal. 2021;39:280. [doi: 10.11604/pamj.2021.39.280.25695]

Available online at: https://www.panafrican-med-journal.com/content/article/39/280/full

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Research

Evaluation of cellular changes in blood stored for transfusion at Bungoma County Referral Hospital, Kenya

Evaluation of cellular changes in blood stored for transfusion at Bungoma County Referral Hospital, Kenya

Phidelis Maruti Marabi1,2,&, Stanslaus Musyoki2, Angela Amayo3

 

1Bungoma County Referral Hospital, Bungoma, Kenya, 2School of Health Sciences, Kisii University, Kisii, Kenya, 3Department of Human Pathology, University of Nairobi, Nairobi, Kenya

 

 

&Corresponding author
Phidelis Maruti Marabi, Bungoma County Referral Hospital, Bungoma, Kenya

 

 

Abstract

Introduction: during the storage of transfusion blood, it may undergo a series of cellular changes that in speculation could be the reason behind the risk of using prolonged stored blood. It's important therefore to monitor the cellular changes that may reduce its survival and function. The objective was to assess the cellular changes in whole blood stored for transfusion at Bungoma county referral hospital.

 

Methods: a prospective study design involving 20 randomly selected donor blood units in citrate phosphate dextrose adenine (CPDA-1) anticoagulant was employed, cellular changes were evaluated for 35 days. The changes were tested using the Celtac F Haematology analyzer. Statistical Analysis of variance was employed in the descriptive statistics. All the investigation was executed using statistical package for social sciences (SPSS V.23). Results were regarded as significant as P < 0.05. Results were presented in tables and charts.

 

Results: at the end of the 35 days blood storage at blood bank conditions, white blood cell (WBC), red blood cell (RBC), platelets counts and mean cell haemoglobin concentration (MCHC) decreased significantly (P < 0.0001, = 0.0182, < 0.0001,= 0.0035). The mean cell volume (MCV), haematocrit (HCT) and mean cell haemoglobin (MCH) increased significantly (P < 0.0001, = 0.0003, = 0.0115) while HGB had insignificant variance (P = 0.4185).

 

Conclusion: platelets, WBC, RBC counts, and indices are significantly altered in stored blood especially when stored over two weeks based on most of the cellular components analyzed in this study. The study, therefore, recommends the utilization of fresh blood to avoid the adverse outcome of cellular changes of reserved blood.

 

 

Introduction    Down

Blood is a composite tissue constituting cell and non-cell elements that perform multiple roles [1]. The non-cell elements comprise the plasma and its derivatives. The cell components are made up of WBCs, PLTs, and RBCs [1]. Blood to be transfused is kept for up to 35 to 42 days at 2-6°C in preservatives such as citrate phosphate dextrose adenine (CPDA) [2]. During storage, blood experience a sequence of cellular changes which minimize their lifespan and purpose, and the most affected product is whole blood [3]. The deterioration in blood and cellular constituents happen almost immediately it is removed from the donor and recipients in need of transfusions rely on the blood and blood components safety and potency [4]. To minimize the dangers linked with blood transfusion, advanced anticoagulants, additive solutions, red blood cell membrane stabilizers, preservatives, and bags were manufactured [5]. Even with these developments, several changes in blood stored for transfusion have been encountered and referred to as red blood cell storage lesions´. After transfusion integral hemodynamic is similar for current and stored cells; nonetheless, microvascular hemodynamic are acutely affected by stored cells which minimizes blood movement and oxygen transport. Furthermore, the existence of stored cells in the bloodstream alter cell-cell and cell-wall interchanges and modify the cell [3]. The oxidative injury manifests red cells extra vulnerable to stress as indicated by increased osmotic fragility in the course of the storage and resultant discharge of haemoglobin (HGB) and intracellular enzymes such as lactate dehydrogenase (LDH) into the floating plasma [6].

Even though RBCs might be stored at 2-6°C for up to forty-two days before transfusion, less is understood of how changes to RBCs in the course of storage might alter their attachment properties [7]. Some studies have shown that stored RBCs show radical deformability changes in the course of blood storage at 2-6°C. Studies have denoted that the distortion index of RBCs does not vary substantially during blood storage at 2-6°C. Nonetheless, radical differences prevail in time constants and circularity distribution widths, which can be utilized to estimate stored red blood cell quality or age [8]. During blood storage at 2-6°C, glycolysis is retarded and, as acid pile up, the amount of ATP reduce and the structure of the RBC is bit-by-bit changed from disc-shaped to echinocytic shapes [9]. The release of unbound haemoglobin following red blood cells lysis during blood storage and its effect on the intravascular nitric oxide metabolism after transfusion has been considered a predominant role [10]. Studies have denoted that transfusion of long-stored blood is linked with a rise in plasma unbound haemoglobin and hunting of nitric oxide in vitro [11]. In line with this discovery, elevated unbound haemoglobin levels in patients with chronic and severe hemolysis have been connected with reduced nitric oxide bioavailability within the micro-capillary bed, reduced organ perfusion, and raised organ injury [12]. Similarly, transfusion of stored blood pints may increase unbound haemoglobin levels in recipients after transfusion - for example, as a result of a pre-mature intravascular burst of the transfused RBCs, or because of the transfusion of the unbound haemoglobin -containing storage medium [13]. Consistent with this hypothesis, studies have shown that transfusing of unbound haemoglobin containing stored blood cause a significant rise of blood pressure in rats that is correlated with the unbound haemoglobin levels in the stored blood [14]. With prolonged storage, there is a shortage of ATP, then the pumps may not be able to maintain the ionic homeostasis of the red blood cell, leading to changes in shape and mean cell volume (MCV), haematocrit (HCT), mean cell haemoglobin (MCH) and mean cell haemoglobin concentration (MCHC). The water influx to the cytosol gives rise to the swelling of erythrocytes during storage and the non-existence of selective channels of performance as done in the spleen are associated with the change in shape and volume of the red blood cell during storage [15].

Studies have revealed that white blood cells have a short life span in stored blood (only a few hours) and transfusion after 24 hours of storage has been proved to be ineffective in raising the WBC count of patients [1]. Many studies have revealed that during storage the total WBC count decreases, and they associate this count reduction to degeneration of the granulocytes [1]. Studies have denoted that WBC depletion in the course of blood storage has been connected with ATP depletion and white blood cells being used in the development of micro-aggregates, which are a mixture of white blood cells, platelets, fibrin, cold insoluble globulin, and cellular debris formed during storage [16]. Studies show that in the course of storage, changes take place in both platelet and storage device, which may lead to the activation of platelet and malfunction [17]. The stimulation of Platelets in the course of storage of blood leads to Platelet-white blood cell aggregates (PLAs) accumulation that introduces WBC apoptosis. Pro-coagulant action, likely correlated with micro-particles from apoptotic white blood cells, might lead to harmful properties of stored blood [18]. Results from some studies indicate that canine platelets survive when stored at room temperature for up to eight hours in CPDA-1 treated whole blood. However, a gap still exists as to whether there are changes after 8 hours of storage [19]. The establishment of blood storing arrangements allows time between collection of blood and transfusion to be increased. This time increase has allowed the decentralization of blood donation utilities with successive savings and advancements in the accessibility of blood components. However, the accessibility of blood storage increases the question of to what extend blood components can and should be stored and to what extent are they safe and potent [20].

Elongated blood storage increases mortality, serious infections, and multi-organ failure after transfusion; however, the causes of these remain unknown [21]. According to records in Bungoma County Referral Hospital, a monthly average of 2-3% of patients reacts to transfused blood especially aged blood ( > 20 days). Blood toxicity has been speculated to be a result of changes as blood ages which are not monitored during storage [22]. Despite this, little is understood about the changes that take place in the course of storage of blood cells during blood storage at Bungoma and at large. This study thus; determined the cellular, biochemical changes, and bacterial contamination in whole blood stored for transfusion so that transfusion safety is ensured. Study objective: this study assessed cellular changes in whole blood stored for transfusion at Bungoma County Referral Hospital in the western region of Kenya between February 2019 and August 2019.

 

 

Methods Up    Down

Ethical considerations: the study was cleared by the Ethical committee of Jaramogi Oginga Teaching and Referral Hospital (#ERC.IB/VOL.1/454) and authority to carry out the research was issued by the National Commission for Science, Technology and Innovation (#NACOSTI/P/19/32125/27143). Written informed consent was obtained from each donor after a brief explanation of what the study is about. Participation in the study was voluntary. Donor details were kept confidential by the exclusion of all forms of identification on the data collection tool and filled data tools were stored physically under lock and digitally with restricted password access on a computer. The donors were informed that their blood units if selected for the study, were not transfused to patients.

Study area: the study was carried out between February and August 2019 at Bungoma County Referral Hospital that is located in Bungoma County (coordinates 0.4213°N to 1.1477° N along the latitude and 34.3627°E to 35.0677°E along the longitude) in Western Kenya. This was a good study area because blood reactions among the patients who receive a blood transfusion were common. The hospital also has an accredited laboratory that is well-equipped with haematology equipment required for the study. The hospital has a blood donation centre that collects an averagely of 600 blood units monthly.

Study design: the research employed a prospective study design and involved collection of blood pints from normal volunteers and reserved under blood bank conditions then tested at baseline and then serially at day 7, day 14, day 21, day 28, and five. The whole unit of blood was used in the study to ensure that the minimum storage condition for blood meant for transfusion was observed.

Study population: the sample size of the research was determined by Yamane Taro formulae for the finite population [23]. A sample size of 20 blood units was adopted for the study. A simple random sampling technique was used in this study where every 10th sample was selected for the study to eliminate bias.

Data collection

Sample collection and analysis

All blood units were collected according to blood transfusion donor guidelines as described by the World Health Organisation [24]. At baseline, samples were immediately separated from blood units collected from the volunteer donors to test for WBC count, RBC count, HGB level, MCV, HCT, MCH, MCHC, and Platelet count. Blood units were then stored at blood bank conditions of 2-6°C for 35 days with intermittent sampling at 7-day intervals to test for White blood cell count, Red blood cell count, and Haemoglobin level, mean cell volume, Haematocrit, Mean Cell Haemoglobin, Mean Cell Haemoglobin Concentration and platelet count. The samples for testing were aseptically transferred from blood units to plain test tubes during separation, No, any other anticoagulant or additives were added to the separated samples. The separated samples were brought to the right temperature as per the manufacturer's instructions before analysis. The laboratory results were recorded on a standard data collection form developed for this study. Cellular changes (Including RBC count, WBC count, and platelet count, HCT, and MCV) and haemoglobin changes were tested using Celtac F MEK-8222 haematology analyzer (NIHON KOHDEN Corporation, 1-31-4 Nishiochiai, Shinjuku-ku, Tokyo 161-8560, Japan). Briefly, the procedure involved pressing numerical on the screen to enter sample identification details; placing well-mixed samples on the autoloader; pressing the start icon on the screen to start sample aspiration and analysis; the results were displayed on the screen and automatically printed immediately the instrument completed the analysis. The normal ranges are White blood cells count: 3.5-10.0 x 109/L; Red blood cells: 4.50-6.50 x 1012/L; Haemoglobin: 12.5-16.5g/dL; Haematocrit: 25.0- 45.0%; Mean cell volume: 70.0 - 100.0 fL; Mean cell haemoglobin concentration: 31.0- 38.0 g/dL; and platelets: 130-400 x109/L.

Quality assurance of the data

To ensure that quality of data collected, pre-donation requirements (such as ensuring that only donors with a haemoglobin level of 12.5g/dL and above, weighing 50 kilograms and above, without a history of recent (12 months) transfusion, that have not recently donated blood (3 months after the last donation), without history and signs of malignancy, without signs and symptoms of sickle cell disease, without signs and symptoms of polycythaemia Rubra Vera and without a history of haemophilia and other coagulation disorders donate blood) were followed and only blood units that met these criteria were used. A qualified phlebotomist collected the blood samples, ensuring that the right quantity of between 450 to 500ml was collected. The pints were transported and stored as per blood transfusion guidelines at 2-8°C. Sample aliquots at the various study points were brought to the optimum temperature as per the manufacturer's instructions before testing. Samples were analyzed in duplicates for each sample and an average was computed to ensure accuracy. Both external and internal quality controls for haematology tests were verified and ensured during the study. Recorded results were verified by second personnel to ensure accuracy. The laboratory is enrolled in the Human Quality assessment services (HUQAS) External Quality Assurance Scheme for haematology scope including WBC count, RBC count, HGB level, MCV, HCT, MCH, MCHC, and Platelet count. The haematology scope is also accredited by the Kenya Accreditation Services (KENAS) which further assured the quality of data collected.

Data management and analysis

The data was stored in Microsoft Excel (Microsoft Corporation, Redmond, Washington, United States). The interrogation was done using the Statistical Package for the Social Sciences (SPSS V.23) (IBM Corporation, Chicago, Illinois, United States). Descriptive statistics (frequencies, mean, and standard deviation) were used to describe the data. The trends of the biochemical changes were shown using line plots. Analysis of variance (ANOVA) was used to establish if there were significant biochemical changes in transfusion blood at baseline and each study point compared to normal reference intervals for 35 days of storage. Findings were considered significant at p < 0.05. Tukey´s Honest Significant Difference test was used to collate all feasible pairs of means. Results were presented as tables and charts.

 

 

Results Up    Down

The white blood cell count demonstrated a decreasing trend from 5.85x109/L ± 0.41 to 2.12 ± 0.14x109/L, RBC count demonstrated a slightly decreasing trend from 5.36x1012//L ± 0.41 to 4.91x1012/L ± 0.40, HGB level demonstrated a slightly increasing trend from 15.47g/dL ± 0.45 to 15.89g/dL ± 0.25 while platelet count demonstrated a decreasing trend from 188.90x109/L ± 19.67 to93.80x109/L±8.26 respectively as shown in Figure 1. The MCV demonstrated an increasing trend of 71.14.25fL ± 1 to 83.21.10fL ± 1; HCT demonstrated a slightly increasing trend of 38.08% ± 2.73 to 44.33% ± 2.49; MCH demonstrated a slightly increasing trend of 27.42pg ± 0.71 to 33.49pg ± 2.17 while MCHC demonstrated a slightly decreasing trend of 41.14g/L ± 1.27 to 36.08g/L ± 1.01 as shown in Figure 2. This study observed that white blood cell count is affected by storage at 2-6°C right from the 14th day of storage as shown in Table 1. Red blood cell count is affected by storage from the 35th day of storage as shown in Table 1. There were no significant changes in haemoglobin level at each testing time point as shown in Table 1. Mean cell volume is affected by storage at 2-6°C from the 7th day of storage as shown in Table 1. Haematocrit is affected by storage at 2-6°C right from the 28th day of storage as shown in Table 1. Mean cell haemoglobin is affected by storage from the 35th day of storage as shown in Table 1. Mean cell haemoglobin concentration is affected by storage from the 35th day of storage as shown in Table 1. While platelets count is affected by storage right from the 14th day as shown in Table 1.

 

 

Discussion Up    Down

The present study has shown that there are cellular changes during the storage period. The white blood cells count changes were non-significant after one week of storage. However significant reduction was observed after 14 days which further decreased significantly through to 35 days of storage. These results, therefore, indicate that: white blood cells during storage are significantly altered by the 7th day. Factors that contribute to the white blood cells reduction during blood for transfusion storage could be loss viability because of ATP depletion. More so, leukocytes are used in the formation of white blood cell- platelet micro-aggregates, which are a mixture of white blood cells, platelets, fibrin, cold globulin, and cellular debris formed during storage (Ahmed, 2008). The clinical significance of this finding is that stored blood for transfusion could be particularly ineffective as a clinical tool in the management of aplastic anaemia and other leucopenic patients since the most critical establishment in these conditions is almost always neutropenia [25]. These findings do compare with findings of a study done in Braithwaite Memorial Specialist Hospital (BMSH), Port Harcourt, Rivers State, Nigeria which demonstrated that at 28 days, there were significant changes in white blood cell differential and absolute counts [26]. These results also concurred with the findings of another study carried out in Aminu Kano Teaching Hospital, Kano, Nigeria which illustrated that the percentage fall from day-zero to day-thirty five was 97% for white blood cells [27]. These findings also concurred with findings of another study done in the Veterinary Transfusion Research Laboratory, 85 University of Milan, Italy which showed that there was a statistically significant drop in WBC count after storage for 35 days of transfusion blood [28]. Another study done in L. N. Medical College and J. K. Hospital, Bhopal, India which demonstrated that WBC count constantly decreased throughout the 28 days storage period also concurred with these findings (Bhargava, Gupta, Vivek, & Khare, 2016). To the strength of the findings from the present study, white blood cells count monitoring during blood for transfusion storage intending to improve blood transfusion efficacy and safety and is recommended.

In the current study, the red blood cells count changes were insignificant up to three weeks (21 days) of storage. However significant reduction was observed at day 28 and further significantly decreased through to 35 days of storage. These results, therefore, indicate that; Red blood cells during storage are significantly altered by the 28th day. The current findings can be explained by the fact that the systemic and biochemical changes that red blood cells go through in the course of storage are anticipated to be instrumental to the drop in red blood cell count as storage span increase [29]. During blood storage at 2-6°C, glycolysis is reduced and as acid level increase, the amount of ATP reduce, and the structure of the red cell is slowly changed from discoid to echinocytic shape [30]. If there is a shortage of ATP, then the pumps (co-transporters) may be unable to maintain the ionic homeostasis of the cell, leading to changes in red blood shape and volume [6]. The number of undamaged red blood cells that remain in a prolonged-stored blood unit before transfusion is not known and warrants additional research. A human red blood cell has a lifecycle of about one hundred and twenty days [31]. In normal conditions, about 2.4 million new red blood cells are generated per second with the concomitant eviction of an equivalent quantity of senescent red blood cells from the bloodstream. Hence, human blood constitutes reds blood cells that vary from zero to one hundred and twenty days of age, which is identical to a pint of freshly collected blood [32]. This experience may likely be suggestive of some level of cell selection where older and more labile cells die initially rapidly, thus leaving a cohort of younger and more stable cells that die later at a much slower rate [27]. The clinical significance of these findings is that adjustment in architecture from basic bio-concave rings to echinocytic red blood cells makes the cells easier to clump, increasing the possibility of blocking the microcirculation, leading to tissue ischemia (Adam, 2015). These less elastic red blood cells are not able to cross tiny micro-vessels of the micro-circulation, leading to reduced oxygen transport since the aerated red cells can't cross the end organ capillary beds (Yalcin, 2014). Transfused stored RBCs can provoke a pro-inflammatory response by the cytokines and eicosanoids. Storage lesions also promote adhesion to endothelial cells, complement system activation, and changes in coagulability. These effects also damage the endothelial lining to cause capillary leakage [9]. The pro-inflammatory nature of stored RBCs has been correlated with an increased fatality, multiple organ failure, thrombosis, and protracted hospital stay [33].

These findings compare with findings from the previous study done in Port Harcourt, Rivers State, Nigeria which showed no statistically significant changes in red blood cell count in the course of the 28 days storage period [26]. These findings also compare with the findings of a study conducted in Bhopal, India which demonstrated no significant changes in red blood cell count during the 28 days storage period [16]. These findings also compare with the findings of a study conducted in Rohtak, India which demonstrated no significant changes in red blood cell count during the 28 days storage period [34]. However, the finding of this study contradicts the findings of a study done in L. N. Medical College and J. K. Hospital, Bhopal, India which showed that RBC count increased during the 28 day storage period (Bhargava et al., 2016). These findings also differ from the findings of a study done in So Joo Hospital, Porto, Portugal which showed that the RBC count kept unchanged throughout the 42 days of storage [5]. These findings also differ from the findings of a study done in Sanjay Gandhi Memorial Hospital, Rewa, India which demonstrated that red blood cell count showed no significant change during the 35 days storage period [25]. In light of the findings from the present study, red blood cells count monitoring during blood for transfusion storage intending to improve blood transfusion efficacy and safety is recommended.

In the current study, haemoglobin level estimation demonstrates an insignificant increase throughout the blood storage period. The slight increase in Haemoglobin level can be explained by the fact that during storage, the by products of glycolytic metabolism, lactic acid, and proteins accrue, which in vivo are readily removed from the bloodstream, remain and give rise to physical changes and cell lysis releasing unbound haemoglobin into plasma [35]. The clinical significance of these findings is that transfusing older pints with unbound haemoglobin has transfusion-related harm, especially for patients who have a history of unbound haemoglobin in their circulation [10]. Unbound haemoglobin may trigger vasoconstrictive, pro-oxidative, and pro-inflammatory events that have transfusion-related harm to the transfused patient [9]. Our findings do compare with a previous study in São João Hospital, Porto, Portugal which showed that the haemoglobin amount remained unvaried in the course of the 42 days of reservation [5]. However, our findings contrast a previous study done in the Department of Pathology, S.S. Medical College Rewa, and India which demonstrated that haemoglobin concentration gradually decreased during the 35-day storage period [25]. In light of the findings from the present study, haemoglobin monitoring during blood for transfusion storage and conditions of patients that might lead to the release of unbound haemoglobin in their circulation need to be considered before indicating a transfusion to improve blood transfusion efficacy and safety and is recommended.

In the current study, MCV demonstrates a significant increasing trend from day0 to day 35 storage period. These findings, therefore, indicate that; MCV is significantly increased during storage. The current findings can be explained by the fact that the water influx to the cytosol giving rise to the swelling of erythrocytes during reservation and the non-existence of selective channels of performance, as done in the spleen, could describe the elevation of the mean cell volume in vitro and successive structural red blood cells changes [36]. The clinical significance of the present findings is that the cell mechanical properties and blood rheology are affected compromising blood hemodynamics, O2 delivery, and the interaction between flowing blood and the vasculature [3]. These findings compare with previously documented findings in So Joo Hospital, Porto, Portugal which demonstrated that MCV elevated notably from day 0 to day 21 and kept steady to the end of the storage period [5]. Our findings also compared with another study done in Iran which demonstrated that MCV increased during storage of blood for transfusion [37]. However, these results are in contrast with the result documented in a study done in Braithwaite Memorial Specialist Hospital (BMSH), city of Port Harcourt, Rivers State, Nigeria. which demonstrated that MCV had insignificant change throughout the 28 days storage period [26]. In consideration of the findings from the present study, MCV monitoring during blood for transfusion storage to improve blood transfusion effectiveness and safety is advocated for.

In the present study, HCT demonstrates a slightly increasing trend, with the significant change being noted from day 14 of the storage period and continues throughout the remaining period of storage time. The current findings can be explained by the fact that the increase in haematocrit reflects the morphological alterations that take place during blood storage (Bosman et al. 2008). The clinical significance of these findings is that increased morphological alteration minimizes the potency of transfused blood by increasing the speed of elimination of transfused cells by the macrophage [2]. These results compare with previously documented findings São João Hospital, Porto, Portugal which demonstrated that HCT increased from day 0 to day 14 and remained stable afterward [5]. Our findings also compare with findings from another study done in Doha, Qatar that demonstrated a significant HCT increase after 35 days of blood storage [30]. However, the current study contrasts the findings of a study done in Obafemi Awolowo University Teaching Hospital Complex, Ile-Ife, Osun State, Nigeria which showed a significant fall of haematocrit [38]. Our findings also contrast findings of a study done conducted in Braithwaite Memorial Specialist Hospital (BMSH), city of Port Harcourt, Rivers State, Nigeria which demonstrated an insignificant change of HCT throughout the 28 days storage period [26]. To the strength of the findings from the present study, HCT monitoring during blood for transfusion storage and use of blood less than 14 days to improve blood transfusion efficacy is advocated for.

In the current study, MCH and MCHC demonstrate insignificant variance from day 0 to day 28; it, however, demonstrates significant variance (increase and decrease respectively) at day 35 of storage time. The changes in MCH and MCHC appear to be the result of a deregulated mechanism of cell volume, which expounds the increase in the volume of the RBCs, the increasing hypocromia, and the anisocytosis. Hence, the decrease in MCHC does not result from a reduction in hemoglobin concentration, but an increase in cell volume [5]. These results compare with those findings documented in a study done in Braithwaite Memorial Specialist Hospital (BMSH), city of Port Harcourt, Rivers State, Nigeria which showed that the MCH and MCHC changes were insignificant during the 28 days of storage [26]. However, the results contrast the findings of a study done in Iran which demonstrated that MCH decreased during the storage period [37]. In light of the findings from the present study, MCH and MCHC monitoring during blood for transfusion storage and use of fresh blood less than 28 days with a focus on improving blood transfusion efficacy is recommended.

In the present study, Platelets demonstrates a significant decrease from day 14 and continues throughout the 35 days storage period. The current findings can be explained by the fact that the cell loose viability owing to ATP depletion in addition to platelet utilization due to micro-aggregates development [27]. The clinical significance of these findings is that this may expose patients to possible decreases in platelets effectiveness as well as likely increases in adverse incidences in addition to transfusion-related sepsis, such as inflammation and/or immune-mediated incidences. Seriously sick patients, including post-cardiac surgery patients and haematology/oncology patients, may be specifically vulnerable to platelets´ adverse incidences because of their pre-transfusion inflammatory state [17]. These results are in comparison with those obtained in a study done in Aminu Kano Teaching Hospital, Kano, Nigeria which demonstrated a constant decrease of platelets throughout the 35 days storage period (Ahmed, 2008). Our findings also compare with those results abstained in a study done in L. N. Medical College and J. K. Hospital, Bhopal, India which demonstrated that platelet decreased significantly during the storage period [16]. The findings also compare with the findings documented in a study done in Nigeria, which demonstrated 86.2% platelet count fall from day 0 to day 28 storage times [27], however, contrast the findings of a study done in Nigeria, which demonstrated insignificant platelet count variance throughout the 28 days storage period [26]. In light of the findings from the present study, platelet count monitoring during blood for transfusion storage to improve blood transfusion efficacy and safety is recommended.

Overall, together with other parameters, cellular changes in stored blood for transfusion should be keenly monitored putting into consideration the patient to be transfused, and the clinical indication of the blood. Limitations: the current study assessed cellular changes in blood for transfusion stored at 2-6°C at a single facility, which may not be representative of the whole country.

 

 

Conclusion Up    Down

Platelets, WBC, RBC counts, and indices are significantly altered in stored blood especially when stored over two weeks based on most of the cellular components analyzed in this study. The clinical consequence of this is that long-reserved whole blood would be distinctly worthless as a clinical tool in the management of blood disorders. Fresh whole blood of not more than two weeks is recommended as a better choice of transfusion of whole blood however blood components may vary depending on the level of changes in each blood parameter or indices.

What is known about this topic

  • Evidence that patients react adversely following transfusion is available;
  • Evidence that red blood cells experience harmful changes during storage is available;
  • Evidence that transfusing older, stored blood might lead to increased fatality; severe infections, multiple organ breakdown, thrombosis, and lengthened hospital stay is available.

What this study adds

  • The study has identified what storage extend is stored blood for transfusion potent and safe;
  • The study has shed light on the need to carefully monitor the cellular changes during blood storage;
  • The study has shed light on the need to transfuse patients who have a history of unbound haemoglobin in their circulation with caution.

 

 

Competing interests Up    Down

The authors declare no competing interests.

 

 

Authors' contributions Up    Down

PMM, SKM, and AA were involved in the study conceptualization, blueprint, data collection, data interrogation, manuscript writing, and editing. All the authors read and approved the manuscript.

 

 

Acknowledgments Up    Down

We sincerely extend our thanks to all contributors whose input made this study successful. We wish to also extend our gratitude to the staff of Bungoma County Referral Hospital laboratory and Bungoma Blood Transfusion satellite for their support during donor recruitment, collection of blood units, and analysis of specimens.

 

 

Table and figures Up    Down

Table 1: cellular variance (change) throughout the blood storage period at Bungoma County Referral Hospital, Kenya, February to August 2019

Figure 1: trend of WBC, RBC, HGB and platelets changes in the progressing storage period of transfusion blood at Bungoma County Referral Hospital, Kenya

Figure 2: trend of red blood indices (MCV, HCT, MCH & MCHC) changes in the progressing storage period of transfusion blood at Bungoma County Referral Hospital, Kenya

 

 

References Up    Down

  1. Nuaimy K. Haematological Changes in Stored Blood. J Educ Sci. 2008;21(4):49-56. Google Scholar

  2. Oyet C, Okongo B, Onyuthi RA, Muwanguzi E. Biochemical changes in stored donor units: implications on the efficacy of blood transfusion. J Blood Med. 2018 Jun 25;9:111-115. PubMed | Google Scholar

  3. Yalcin O, Ortiz D, Tsai AG, Johnson PC, Cabrales P. Microhemodynamic aberrations created by transfusion of stored blood. Transfusion. 2014 Apr;54(4):1015-27. PubMed | Google Scholar

  4. Isbister J. Is the Clinical Significance of Blood Storage Lesions Underestimated? Transfus Altern Transfus Med. 2003;5(3):356-362. PubMed | Google Scholar

  5. Diana Noguira EC, Susana Rocha, Estela Abreu. Biochemical and Cellular Changes in Leukocyte-Depleted Red Blood Cells Stored for TransfusionNo Title. Transfus Med Hemotherapy. 2015 Jan;42(1):46-51. PubMed | Google Scholar

  6. Orlov K. The pathophysiology and consequences of red blood cell storage. Anaesthesia. 2015 Jan;70 Suppl 1:29-37, e9-12. PubMed

  7. Anniss AM, Sparrow RL. Storage duration and white blood cell content of red blood cell (RBC) products increases adhesion of stored RBCs to endothelium under flow conditions. Transfusion. 2006 Sep;46(9):1561-7. PubMed | Google Scholar

  8. Zheng Y, Chen J, Cui T, Shehata N, Wang C, Sun Y. Characterization of red blood cell deformability change during blood storage. Lab Chip. 2014 Feb 7;14(3):577-83. PubMed | Google Scholar

  9. Yoshida P, Michel P, Angelo D. Red blood cell storage lesion: causes and potential clinical consequences. Blood Transfus. 2019 Jan;17(1):27-52. PubMed | Google Scholar

  10. Iris Vermeulen CW, de Wit Norbert CJ, Sertorio Jonas TC. Blood transfusions increase circulating plasma free hemoglobin levels and plasma nitric oxide consumption: a prospective observational pilot study. Crit Care. 2012 May 25;16(3):R95. Google Scholar

  11. Simon Finney J. Free haemoglobin in ‘old’ transfused blood - baddy or bystander? Crit Care. 2012 Jul 31;16(4):141. PubMed | Google Scholar

  12. Christopher Reiter D, Xunde Wang, Tanus-Santos JE, Neil Hogg, Richard Cannon O, Alan Schechter N et al. Cell-free hemoglobin limits nitric oxide bioavailability in sickle-cell disease. Nat Med. 2002 Dec;8(12):1383-9 Epub 2002 Nov 11. PubMed | Google Scholar

  13. Mark Gladwin T, Daniel Kim-Shapiro B. Storage lesion in banked blood due to hemolysis-dependent disruption of nitric oxide homeostasis. Curr Opin Hematol. 2009 Nov;16(6):515-23. PubMed | Google Scholar

  14. Donadee C, Nicolas Raat JC, Tamir Kanias, Jesus Tejero, Janet Lee S, Eric Kelley E et al. Nitric oxide scavenging by red blood cell microparticles and cell-free hemoglobin as a mechanism for the red cell storage lesion. Circulation. 2011 Jul 26;124(4):465-76 Epub 2011 Jul 11. PubMed | Google Scholar

  15. Giel Bosman JCGM, Edwin Lasonder, Yvonne Groenen-Dpp AM, Frans Willekens LA, Jan Werre M. The proteome of erythrocyte-derived microparticles from plasma: new clues for erythrocyte aging and vesiculation. J. proteomic. 2012 Dec 5;76 Spec No:203-10. PubMed | Google Scholar

  16. Bhargava P, Gupta R, KhareV. CPDA-1 Stored Blood Induced Effect on Hematological and Biochemical Parameter up to 28 DaysNo Title. Adv path lab Med. 2016;8(12):8-12. PubMed | Google Scholar

  17. Cécile Aubron, Andrew Flint WJ, Yves Ozier, Zoe M. Platelet storage duration and its clinical and transfusion outcomes: a systematic review. Crit Care. 2018 Aug 5;22(1):185. PubMed | Google Scholar

  18. Keating FK, Butenas S, Fung MK, Schneider DJ. Platelet-white blood cell (WBC) interaction, WBC apoptosis, and procoagulant activity in stored red blood cells. Transfusion. 2011 May;51(5):1086-95. PubMed | Google Scholar

  19. Crowther M, Ford I, Jeffrey RR, Urbaniak SJ, and Greaves M. Quality of harvested autologous platelets compared with stored donor platelets for use after cardiopulmonary bypass procedures. 2000 Oct;111(1):175-81. PubMed | Google Scholar

  20. Zimrin AB, Hess JR. Current issues relating to the transfusion of stored red blood cells. Vox Sanguinis. 2009 Feb;96(2):93-103. PubMed | Google Scholar

  21. Sheppard CA, Cassandra Josephson D, Hillyer CD. Bacterial Contamination of Platelets for Transfusion: Recent Advances and Issues. LABMEDICINE. 2005;36(12):767-770. PubMed | Google Scholar

  22. Patrick Burger, Elena Kostova, Esther Bloem, Petra H-S, Alexander BM, Timo van den Berg K et al. Potassium leakage primes stored erythrocytes for phosphatidylserine exposure and shedding of pro-coagulant vesicles. Br J Haematol. 2013 Feb;160(3):377-86. PubMed | Google Scholar

  23. Israel GD. Determining Sample Size. Florida Coop Extention Serv Fact Sheet PEOD-6. 1992. Google Scholar

  24. WHO. Blood donor selection: guidelines on assessing donor suitability for blood donation. Blood Donor Sel. 2012. Google Scholar

  25. Batham N. Evaluation of haematological parameter in stored CPDA-1 whole blood. Int J Appl Res. 2018;4(11):220-223.

  26. BM-I, Teddy Adias ZJ. Storage Related Haematological and Biochemical Changes of CPDA-1 Whole Blood in a Resource-Limited Setting. J Blood Disord Transfus. 2012; 3(3).

  27. Ahmed SG, Orakah JA. Cellular Changes in Stored Whole Blood and the Implication on Efficacy of Transfusion Therapy in Nigeria. Internet J. Third World Med. 2008;8(2). Google Scholar

  28. Eva Spada, Roberta Perego, Luciana Baggiani, Daniela Proviebo. No Haematological and morphological evaluation of feline whole blood units collected for transfusion purposes Title J. Feline Med. Surgery. 2019 Aug;21(8):732-740. PubMed | Google Scholar

  29. Maria García-Roa, Maria Del Carmen Vicente-Ayuso, Alejandro Bobes M, Alexandra Pedraza C, Ataulfo G-F, Maria PM. Red blood cell storage time and transfusion: current practice, concerns and future perspectives. Blood Transfus. 2017 May;15(3):222-231. PubMed | Google Scholar

  30. Mustafa I, Marwani AA, Mamdouh Nasr K, Abdulla Kano N, Tameen H. Time-Dependent Assessment of Morphological Changes: Leukodepleted Packed Red Blood Cells Stored in SAGM. Biomed Res Int. 2016;2016:4529434. PubMed | Google Scholar

  31. Clemente Fernandez Arias, Cristina Fernandez Arias. How do red blood cells know when to die? R Soc Open Sci. 2017 Apr 5;4(4):160850. PubMed | Google Scholar

  32. Wei-Wei Tuo, Wen-Jing Liang, Yao-Xiong Huang. How Cell Number and Cellular Properties of Blood-Banked Red Blood Cells of Different Cell Ages Decline during Storage. PLoS One. 2014 Aug 28;9(8):e105692. PubMed | Google Scholar

  33. Eldad Hod A, Spitalnik SL. Harmful effects of transfusion of older stored red blood cells: Iron and inflammation. Transfusion. 2011 Apr;51(4):881-5. PubMed | Google Scholar

  34. Sonia Chhabra MG, Saurav Chaudhary, Sehgal PK, Sunita Singh, Sen R. Changes in RBC and Platelet indices in CPDA stored BLOOD. Int J Heal care Biomed Res. 2017;5(4):69-75. Google Scholar

  35. Sayeedul Hasan Arif, Nera Yadav, Suhailur Rehman, Ghazala Mehdi. Study of Hemolysis During Storage of Blood in the Blood Bank of a Tertiary Health Care Centre. Indian J Hematol blood Transfus. 2017 Dec;33(4):598-602. PubMed | Google Scholar

  36. Shohag M, Mohammad Raguib Munif, Nargis Jahan, Alam R. Haematobiochemical changes of ovine (ovis aries) blood during storage for transfusion. Res Agric Fish. 2020;7(1):113-120. Google Scholar

  37. Ghezelbash B, Azarkeivan A, Pourfathollah AA, Deyhim M, Hajati E, Alireza G. Comparative Evaluation of Biochemical and Hematological Parameters of Pre-Storage Leukoreduction during RBC Storage. Hematol Oncol Stem Cell Res. 2018 Jan 1;12(1):35-42. PubMed | Google Scholar

  38. Oluyombo R, Oluyombo O, Uchegbwu OO, Adegbamigbe O, Ayodele OE. Quantitative assessment of erythrocytes and leucocytes in CPD- A stored blood. Biomed Res. 2013;24(4):503-508. Google Scholar

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Research

Evaluation of cellular changes in blood stored for transfusion at Bungoma County Referral Hospital, Kenya

Research

Evaluation of cellular changes in blood stored for transfusion at Bungoma County Referral Hospital, Kenya

Research

Evaluation of cellular changes in blood stored for transfusion at Bungoma County Referral Hospital, Kenya