In Situ Monitoring of Morphology Changes and Oxygenation State of Human Erythrocytes During Surfactant-Induced Hemolysis

Erythrocytes, the most abundant blood cells, are a prevalent cell model for the analysis of the membrane-damaging effects of different molecules, including drugs. In response to stimuli, erythrocytes can change their morphology, e.g., shape or volume, which in turns influences their main function to...

Full description

Saved in:
Bibliographic Details
Main Authors: Miroslav Karabaliev, Boyana Paarvanova, Gergana Savova, Bilyana Tacheva, Radostina Georgieva
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Cells
Subjects:
Online Access:https://www.mdpi.com/2073-4409/14/7/469
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849738565131436032
author Miroslav Karabaliev
Boyana Paarvanova
Gergana Savova
Bilyana Tacheva
Radostina Georgieva
author_facet Miroslav Karabaliev
Boyana Paarvanova
Gergana Savova
Bilyana Tacheva
Radostina Georgieva
author_sort Miroslav Karabaliev
collection DOAJ
description Erythrocytes, the most abundant blood cells, are a prevalent cell model for the analysis of the membrane-damaging effects of different molecules, including drugs. In response to stimuli, erythrocytes can change their morphology, e.g., shape or volume, which in turns influences their main function to transport oxygen. Membrane active molecules can induce hemolysis, i.e., release of hemoglobin into the blood plasma. Free hemoglobin in the blood circulation is toxic causing serious health problems including vasoconstriction, high blood pressure and kidney damage. Therefore, early recognition of the risk of massive hemolysis is highly important. Here, we investigated surfactant induced hemolysis applying UV–vis spectrophotometry. Saponin, sodium dodecyl sulfate and Triton X-100, detergents known to provoke hemolysis at different concentrations and by different mechanisms, were applied to initiate the process. Whole absorption spectra of erythrocyte suspensions in the range 300–750 nm were recorded every 15 s for following the process in real-time. The hemolysis process, with respect to morphological changes in the erythrocytes and their influence on the oxygenation state of hemoglobin, was characterized by the absorbance at 700 nm, the height relative to the background and the wavelength of the Soret peak. The results suggest that these UV–vis spectrophotometry parameters provide reliable information in real-time; not only about the process of hemolysis itself, but also about pre-hemolytic changes in the erythrocytes, even at sub-hemolytic surfactant concentrations.
format Article
id doaj-art-5dd8466089d7483fb081237dbd630ab5
institution DOAJ
issn 2073-4409
language English
publishDate 2025-03-01
publisher MDPI AG
record_format Article
series Cells
spelling doaj-art-5dd8466089d7483fb081237dbd630ab52025-08-20T03:06:31ZengMDPI AGCells2073-44092025-03-0114746910.3390/cells14070469In Situ Monitoring of Morphology Changes and Oxygenation State of Human Erythrocytes During Surfactant-Induced HemolysisMiroslav Karabaliev0Boyana Paarvanova1Gergana Savova2Bilyana Tacheva3Radostina Georgieva4Department of Physics and Biophysics, Faculty of Medicine, Trakia University, 11 Armeiska, 6000 Stara Zagora, BulgariaDepartment of Physics and Biophysics, Faculty of Medicine, Trakia University, 11 Armeiska, 6000 Stara Zagora, BulgariaDepartment of Physics and Biophysics, Faculty of Medicine, Trakia University, 11 Armeiska, 6000 Stara Zagora, BulgariaDepartment of Physics and Biophysics, Faculty of Medicine, Trakia University, 11 Armeiska, 6000 Stara Zagora, BulgariaDepartment of Physics and Biophysics, Faculty of Medicine, Trakia University, 11 Armeiska, 6000 Stara Zagora, BulgariaErythrocytes, the most abundant blood cells, are a prevalent cell model for the analysis of the membrane-damaging effects of different molecules, including drugs. In response to stimuli, erythrocytes can change their morphology, e.g., shape or volume, which in turns influences their main function to transport oxygen. Membrane active molecules can induce hemolysis, i.e., release of hemoglobin into the blood plasma. Free hemoglobin in the blood circulation is toxic causing serious health problems including vasoconstriction, high blood pressure and kidney damage. Therefore, early recognition of the risk of massive hemolysis is highly important. Here, we investigated surfactant induced hemolysis applying UV–vis spectrophotometry. Saponin, sodium dodecyl sulfate and Triton X-100, detergents known to provoke hemolysis at different concentrations and by different mechanisms, were applied to initiate the process. Whole absorption spectra of erythrocyte suspensions in the range 300–750 nm were recorded every 15 s for following the process in real-time. The hemolysis process, with respect to morphological changes in the erythrocytes and their influence on the oxygenation state of hemoglobin, was characterized by the absorbance at 700 nm, the height relative to the background and the wavelength of the Soret peak. The results suggest that these UV–vis spectrophotometry parameters provide reliable information in real-time; not only about the process of hemolysis itself, but also about pre-hemolytic changes in the erythrocytes, even at sub-hemolytic surfactant concentrations.https://www.mdpi.com/2073-4409/14/7/469erythrocyteshemolysissaponinSDSTriton X-100hemoglobin
spellingShingle Miroslav Karabaliev
Boyana Paarvanova
Gergana Savova
Bilyana Tacheva
Radostina Georgieva
In Situ Monitoring of Morphology Changes and Oxygenation State of Human Erythrocytes During Surfactant-Induced Hemolysis
Cells
erythrocytes
hemolysis
saponin
SDS
Triton X-100
hemoglobin
title In Situ Monitoring of Morphology Changes and Oxygenation State of Human Erythrocytes During Surfactant-Induced Hemolysis
title_full In Situ Monitoring of Morphology Changes and Oxygenation State of Human Erythrocytes During Surfactant-Induced Hemolysis
title_fullStr In Situ Monitoring of Morphology Changes and Oxygenation State of Human Erythrocytes During Surfactant-Induced Hemolysis
title_full_unstemmed In Situ Monitoring of Morphology Changes and Oxygenation State of Human Erythrocytes During Surfactant-Induced Hemolysis
title_short In Situ Monitoring of Morphology Changes and Oxygenation State of Human Erythrocytes During Surfactant-Induced Hemolysis
title_sort in situ monitoring of morphology changes and oxygenation state of human erythrocytes during surfactant induced hemolysis
topic erythrocytes
hemolysis
saponin
SDS
Triton X-100
hemoglobin
url https://www.mdpi.com/2073-4409/14/7/469
work_keys_str_mv AT miroslavkarabaliev insitumonitoringofmorphologychangesandoxygenationstateofhumanerythrocytesduringsurfactantinducedhemolysis
AT boyanapaarvanova insitumonitoringofmorphologychangesandoxygenationstateofhumanerythrocytesduringsurfactantinducedhemolysis
AT gerganasavova insitumonitoringofmorphologychangesandoxygenationstateofhumanerythrocytesduringsurfactantinducedhemolysis
AT bilyanatacheva insitumonitoringofmorphologychangesandoxygenationstateofhumanerythrocytesduringsurfactantinducedhemolysis
AT radostinageorgieva insitumonitoringofmorphologychangesandoxygenationstateofhumanerythrocytesduringsurfactantinducedhemolysis