Blood Bag Plasticizers Influence Red Blood Cell Vesiculation Rate without Altering the Lipid Composition of the Vesicles

Abstract

Background: Polyvinyl chloride (PVC) plasticized with di(2-ethylhexyl) phthalate (DEHP) is commonly used for blood collection and storage. DEHP has protective effects on RBC membranes, but is also a toxin. Methods: A paired study was conducted to investigate the influence of DEHP and two alternative plasticizers, 1,2-cyclohexane-dicarboxylic acid diisononyl ester (DINCH) and n-butyryl-tri-n-hexyl citrate (BTHC), on the preservation of RBCs stored for 42 days in PVC pediatric bags. The RBC membrane was evaluated for supernatant hemoglobin (Hb), release of extracellular microvesicles (EVs), osmotic fragility, deformability, and lipid composition. Results: In BTHC-plasticized bags, the supernatant Hb increase during storage was 2 times greater than in DINCH- and DEHP-plasticized bags. By day 21, EV concentrations had doubled from day-5 levels in DINCH- and DEHP-, and trebled in BTHC-plasticized bags. RBC mean cell volumes were greater in BTHC- than in DINCH- or DEHP-plasticized bags (p < 0.001). Osmotic fragility differed significantly among plasticizers (p < 0.01). After day 21, RBC deformability decreased in all, but to a greater extent in the bags with BTHC. Phospholipid composition of RBCs and EVs was not different among plasticizers. Conclusion: Membrane stabilization capacity differed among the plasticizers. RBC in BTHC bags stored more poorly, while DEHP and DINCH bags offered better protection against vesiculation, osmotic stress, and Hb loss.

Phospholipid composition of packed red blood cells and that of extracellular vesicles show a high resemblance and stability during storage

“…The strengths of our study: We analyzed the four major PLs of both RBCs and EVs derived from the same RBC concentrates from early, middle and late time points of storage within the Finnish clinical usage period to enable a rational comparison. Species level PL analysis of packed RBCs has been done only by one other research group with partly conflicting results [27,28]. In our study, several appropriate MS scans were run for each type of lipid to obtain reliable results not biased by adducts and other contaminants.…”

“…The lipid composition of the erythrocyte membrane is thoroughly characterized at the PL class level as the erythrocyte has been used as a model cell in biomembrane lipidomic studies [23][24][25]. PL composition at the molecular species level has been less studied [26][27][28], and a storage time-dependent study on RBCs and corresponding EVs is lacking. Accordingly, we aimed to study the time course of changes in PL classes and species of packed RBCs and the corresponding EVs during a clinically relevant storage period.…”

The Blood Bag Plasticizer Di-2-Ethylhexylphthalate Causes Red Blood Cells to Form Stomatocytes, Possibly by Inducing Lipid Flip-Flop

“…The DEHP is known to have a beneficial effect on RBCs that are stored for transfusion, reducing susceptibility to hemolysis [8,9,10,29] and minimizing the formation of both echinocytes [10] and microparticles [8,9,10,11]. Hemolysis, vesicle formation, and the shape changes associated with echinocyte formation are associated with the membrane; together with the known association of DEHP with the RBC membrane [3], this makes the DEHP-membrane interactions a specific target to study.…”

“…Storage materials with alternative plasticizers have been investigated for RCCs, due to the potential health effects of DEHP [8,9,10,11,12]. One challenge associated with the replacement of DEHP is that it has beneficial effects on the RBCs, extending the shelf life of the stored RCCs [8,9,10].…”

“…One challenge associated with the replacement of DEHP is that it has beneficial effects on the RBCs, extending the shelf life of the stored RCCs [8,9,10]. The DEHP reduces RBC vesiculation and loss of hemoglobin [9] and also reduces the extent of morphological changes that occur during storage [10]. …”