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Synergistic Effect of Trehalose and Saccharose Pretreatment on Maintenance of Lyophilized Human Red Blood Cell Quality
Abstract
Purpose: To investigate the synergistic effect of trehalose and saccharose pretreatment on maintenance of lyophilized human red blood cell (RBC) quality.
Methods: RBCs were pre-treated with trehalose and saccharose, and then lyophilized and re-hydrated. Prior to lyophilization and after re hydration, RBC parameters, RBC counts, total hemoglobin concentration, mean corpuscular hemoglobin (MCH), mean corpuscular volume (MCV), comprehensive deformation index, hemolysis ratio and phosphatidylserine (PS) expression, were determined using a hematology analyzer, an RBC deformation instrument, a spectrophotometer and a flow cytometer, respectively. Superoxide dismutase (SOD), glucose-6-phosphate dehydrogenase (G-6-PD), and adenosine triphosphatase (ATPase) activities were determined using kits for SOD, ATPase, and G-6- PD assay, respectively.
Results: After lyophilization-rehydration, RBC counts and total hemoglobin recovery rates, deformability, and RBC SOD, ATPase, and G-6-PD activities were significantly decreased by 47.24 – 74.65 % (p < 0.01), compared with the normal group. RBC osmotic fragility and PS expression on the outer surface of the RBC membrane were significantly increased by 168.53 and 629.30 % (p < 0.01), respectively, compared with the normal group. RBC MCH and MCV values were not significantly affected by lyophilization rehydration (p > 0.05). Trehalose and saccharose pretreatment significantly reversed the effects of lyophilization-rehydration on these RBC parameters by approximately 13.16 – 211.11 % (p < 0.01), compared with the control group. The combined effects were synergistic.
Conclusion: Trehalose and saccharose pretreatment synergistically enhances maintenance of lyophilized RBC quality.
Keywords: Trehalose, Saccharose, Lyophilization, Red blood cell, Hematological parameters