The Platelet Adhesion Capacity to Subendothelial Matrix and Collagen in a Flow Model during Storage of Platelet Concentrates for 7 Days

Abstract

SummaryThe influence of storage of platelet concentrates (PC) on the adhesion capacity of platelets was studied. Twenty-four PC, 12 prepared by the buffy coat (BC) method and 12 by the platelet-rich plasma (PRP) method, were stored for 7 days at room temperature. On days 1,3 and 7 of storage, the platelet adhesion capacity to subendothelial matrix (SEM) and collagen was studied in a rectangular perfusion system under flow conditions in conjunction with the platelet aggregation capacity after stimulation and the adenine nucleotide content. The platelet adhesion capacity to collagen was constant until day 3 of storage and decreased to about 80% of the starting value on day 7 of storage. The adhesion capacity to SEM, however, had already decreased on day 3 to about 75% of the value of day 1 and was even more decreased on day 7 to about 45% of the starting value. On day 1, platelets prepared by the BC method displayed a higher adhesion capacity to collagen and a higher aggregation capacity after stimulation by collagen alone or in combination with ADP, compared to platelets prepared by the PRP method. No other significant differences in adhesion or aggregation capacity were observed between the PC prepared by the two different methods. Both platelet adhesion and aggregation response decreased during storage, as did the total adenine nucleotide content. This study shows that platelet function, as measured by the aggregation and adhesion capacity, of platelets prepared by the PRP method is more severely impaired during the first 3 days of storage as compared to the function of platelets prepared by the BC method.

Platelet adhesion changes during storage studied with a novel method using flow cytometry and protein-coated beads

“…The collagen beads did not change in percentage of beads with adhering platelets, but did increase the amount of platelets adhering to each bead during the first 7 days of storage. There are few studies on platelet adhesion during storage and they vary in the method of platelet preparation, storage length and storage media, making direct comparisons difficult, as all these factors affect platelet function [6,7,[9][10][11]. Furthermore these studies have tested platelets in presence of added red blood cells which could also influence their results.…”

“…One reason is that current available methods are time consuming, complicated to use, and require large sample volumes. The adhesion capacity of stored platelets has been studied using perfusion chambers [6][7][8] or cone and-plate devices [9,10]. These methods require the presence of red blood cells, which complicates the analysis of PCs and introduces a potential source of non-specific and unpredictable effects [8,9,11].…”

Biochemistry of storage lesions of red cell and platelet concentrates: A continuous fight implying oxidative/nitrosative/phosphorylative stress and signaling

“…The intracellular imbalance between GSH and GSSH occurring during storage may also influence the oxidation state of the unpaired thiols present in the cytoplasmic tails of GPVI platelet receptors, presumably impairing the disulfide-dependent dimerization of GPVI which is known to occur during PLT activation [81]. After all stored PLTs progressively lose their adhesive capacity [82]. As specific oxidative (but also nitrosative) modifications of reactive thiol groups are expected to modulate the function of both PLT surface and cytoplasmic proteins, their investigation by proteomics tools may facilitate a more mechanistic understanding of PSLs.…”

Comparative Studies of a Humanized Anti-glycoprotein IIb/IIIa Monoclonal Antibody, YM337, and Abciximab on in Vitro Antiplatelet Effect and Binding Properties.

“…Platelet Adhesion Assay Human von Willebrand factor (vWF) was purified from human plasma by the methods of Thorell et al 15) Subendothelial matrix was prepared by the methods of Boomgaard et al 16) HUVEC (Kurabo Industries, Osaka, Japan) were cultured to subconfluent, and then re-cultured for 2 d in 96 well plates (Nunc, Roskilde, Denmark) for tissue culture. After treatment of the plate with phosphatebuffered saline (PBS) containing 0.5% Triton X-100 and 20 mM NH 4 OH for 3 min at room temperature, the plate was washed 4 times with PBS to obtain a preparation of subendothelial matrix.…”