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Hemodilution Increases the Susceptibility of Red Blood Cells to Mechanical Shear Stress During In Vitro Hemolysis Testing

Christian Sargent, Ina Laura Perkins, Venkat Kanamarlapudi Orcid Logo, Christopher Moriarty, Sabrina Ali

ASAIO Journal, Volume: 67, Issue: 6, Pages: 632 - 641

Swansea University Authors: Christian Sargent, Venkat Kanamarlapudi Orcid Logo

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Abstract

The American Society for Testing and Materials (ASTM) F1841 standard for the assessment of hemolysis in blood pumps recommends using phosphate-buffered saline (PBS) for hemodilution to standardize hematocrit (HCT). However, PBS increases red blood cell mechanical fragility and hemolysis. Herein, we...

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Published in: ASAIO Journal
ISSN: 1058-2916
Published: Ovid Technologies (Wolters Kluwer Health) 2021
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URI: https://cronfa.swan.ac.uk/Record/cronfa57577
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spelling 2022-10-27T11:21:57.5875420 v2 57577 2021-08-10 Hemodilution Increases the Susceptibility of Red Blood Cells to Mechanical Shear Stress During In Vitro Hemolysis Testing 949eccfb07970edf2941844b67a29d1a Christian Sargent Christian Sargent true false 63741801137148abfa4c00cd547dcdfa 0000-0002-8739-1483 Venkat Kanamarlapudi Venkat Kanamarlapudi true false 2021-08-10 The American Society for Testing and Materials (ASTM) F1841 standard for the assessment of hemolysis in blood pumps recommends using phosphate-buffered saline (PBS) for hemodilution to standardize hematocrit (HCT). However, PBS increases red blood cell mechanical fragility and hemolysis. Herein, we investigated diluents and dilutions during in vitro testing to reduce hemodilution bias when assessing hemolysis. Bovine blood was diluted with either PBS or PBS + 4/6 g% bovine serum albumin (BSA) to a 70/90% blood dilution, or to an HCT of 30% ± 2%, and pumped with the CentriMag or RotaFlow under hemodynamic conditions. Separately, bovine and human blood were subjected to ventricular assist device–like shear stress using a vortex. Plasma-free hemoglobin levels, normalized milligram index of hemolysis (mgNIH), and protein concentrations were analyzed. Hemolysis depended on the diluent and final blood concentration. Seventy percent of blood diluted with PBS alone caused significantly greater hemolysis than PBS + 4/6 g% BSA. However, at 90% blood, PBS + 4/6 g% BSA caused significantly greater hemolysis than PBS alone. Hence, a positive correlation between mgNIH and hemodilution was observed with PBS and a negative correlation with PBS + 4g% BSA. PBS alone significantly reduced the total protein concentration. Hemodilution with BSA maintains protein concentration within a physiologic range and reduces bias during hemolysis testing at high blood dilutions. Thus, American Society for Testing and Materials standards could consider including BSA as a diluent, when and as required: where large dilution is required (<83%) use PBS + 4 g% BSA, otherwise use PBS alone. Journal Article ASAIO Journal 67 6 632 641 Ovid Technologies (Wolters Kluwer Health) 1058-2916 1 6 2021 2021-06-01 10.1097/mat.0000000000001280 COLLEGE NANME COLLEGE CODE Swansea University 2022-10-27T11:21:57.5875420 2021-08-10T11:29:58.9911415 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine Christian Sargent 1 Ina Laura Perkins 2 Venkat Kanamarlapudi 0000-0002-8739-1483 3 Christopher Moriarty 4 Sabrina Ali 5
title Hemodilution Increases the Susceptibility of Red Blood Cells to Mechanical Shear Stress During In Vitro Hemolysis Testing
spellingShingle Hemodilution Increases the Susceptibility of Red Blood Cells to Mechanical Shear Stress During In Vitro Hemolysis Testing
Christian Sargent
Venkat Kanamarlapudi
title_short Hemodilution Increases the Susceptibility of Red Blood Cells to Mechanical Shear Stress During In Vitro Hemolysis Testing
title_full Hemodilution Increases the Susceptibility of Red Blood Cells to Mechanical Shear Stress During In Vitro Hemolysis Testing
title_fullStr Hemodilution Increases the Susceptibility of Red Blood Cells to Mechanical Shear Stress During In Vitro Hemolysis Testing
title_full_unstemmed Hemodilution Increases the Susceptibility of Red Blood Cells to Mechanical Shear Stress During In Vitro Hemolysis Testing
title_sort Hemodilution Increases the Susceptibility of Red Blood Cells to Mechanical Shear Stress During In Vitro Hemolysis Testing
author_id_str_mv 949eccfb07970edf2941844b67a29d1a
63741801137148abfa4c00cd547dcdfa
author_id_fullname_str_mv 949eccfb07970edf2941844b67a29d1a_***_Christian Sargent
63741801137148abfa4c00cd547dcdfa_***_Venkat Kanamarlapudi
author Christian Sargent
Venkat Kanamarlapudi
author2 Christian Sargent
Ina Laura Perkins
Venkat Kanamarlapudi
Christopher Moriarty
Sabrina Ali
format Journal article
container_title ASAIO Journal
container_volume 67
container_issue 6
container_start_page 632
publishDate 2021
institution Swansea University
issn 1058-2916
doi_str_mv 10.1097/mat.0000000000001280
publisher Ovid Technologies (Wolters Kluwer Health)
college_str Faculty of Medicine, Health and Life Sciences
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hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
document_store_str 0
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description The American Society for Testing and Materials (ASTM) F1841 standard for the assessment of hemolysis in blood pumps recommends using phosphate-buffered saline (PBS) for hemodilution to standardize hematocrit (HCT). However, PBS increases red blood cell mechanical fragility and hemolysis. Herein, we investigated diluents and dilutions during in vitro testing to reduce hemodilution bias when assessing hemolysis. Bovine blood was diluted with either PBS or PBS + 4/6 g% bovine serum albumin (BSA) to a 70/90% blood dilution, or to an HCT of 30% ± 2%, and pumped with the CentriMag or RotaFlow under hemodynamic conditions. Separately, bovine and human blood were subjected to ventricular assist device–like shear stress using a vortex. Plasma-free hemoglobin levels, normalized milligram index of hemolysis (mgNIH), and protein concentrations were analyzed. Hemolysis depended on the diluent and final blood concentration. Seventy percent of blood diluted with PBS alone caused significantly greater hemolysis than PBS + 4/6 g% BSA. However, at 90% blood, PBS + 4/6 g% BSA caused significantly greater hemolysis than PBS alone. Hence, a positive correlation between mgNIH and hemodilution was observed with PBS and a negative correlation with PBS + 4g% BSA. PBS alone significantly reduced the total protein concentration. Hemodilution with BSA maintains protein concentration within a physiologic range and reduces bias during hemolysis testing at high blood dilutions. Thus, American Society for Testing and Materials standards could consider including BSA as a diluent, when and as required: where large dilution is required (<83%) use PBS + 4 g% BSA, otherwise use PBS alone.
published_date 2021-06-01T04:13:26Z
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