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Transformation of membrane nanosurface of red blood cells under hemin action.

Kozlova E, Chernysh A, Moroz V, Gudkova O, Sergunova V, Kuzovlev A - Sci Rep (2014)

Bottom Line: The process of formation of "grains" was dependent on the hemin concentration and incubation time.The possible mechanism of membrane nanostructure alterations is proposed.This research can be used to study the cell intoxication and analyze the action of various agents on RBC membranes.

View Article: PubMed Central - PubMed

Affiliation: V.A. Negovsky Scientific Research Institute of General Reanimatology RAS, Moscow, Russian Federation.

ABSTRACT
Hemin is the product of hemoglobin oxidation. Some diseases may lead to a formation of hemin. The accumulation of hemin causes destruction of red blood cells (RBC) membranes. In this study the process of development of topological defects of RBC membranes within the size range from nanoscale to microscale levels is shown. The formation of the grain-like structures in the membrane ("grains") with typical sizes of 120-200 nm was experimentally shown. The process of formation of "grains" was dependent on the hemin concentration and incubation time. The possible mechanism of membrane nanostructure alterations is proposed. The kinetic equations of formation and transformation of small and medium topological defects were analyzed. This research can be used to study the cell intoxication and analyze the action of various agents on RBC membranes.

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Stages of transformation of RBC shape and its membrane under the hemin action for different concentrations C.(a) C = 0 (control), AFM 3D- images 100 × 100 μm and 30 × 30 μm of cells in control smear, 95 ± 2% discocytes. (b) C = 0.8 mM, AFM 3D-images 30 × 30 μm, 85 ± 5% deformed discocytes and stomatocytes. (c) C = 1.2 mM, AFM 3D-images 30 × 30 μm, 78 ± 6% planocytes and rising of topological “grain” defects. (d) C = 1.5 mM, AFM 3D-images 30 × 30 μm, 84 ± 10% cells with “grains”-containing domains on the membrane surface. (e) C = 2.5 mM, AFM 3D-images 30 × 30 μm, 79 ± 8% cells with merging domains and spheroechinocytes. Time of incubation was 60 min for (a–e). (f) The dependence of domains arising on incubation time ti for the same hemin concentration C = 1.5 mM (t1 = 3 min, t2 = 60 min and t3 = 180 min). Statistical processing performed by 100 × 100 μm AFM 3D-images of cells in monolayers in 45 smears. (60–100 cells in each image) for each concentration and each time.
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f1: Stages of transformation of RBC shape and its membrane under the hemin action for different concentrations C.(a) C = 0 (control), AFM 3D- images 100 × 100 μm and 30 × 30 μm of cells in control smear, 95 ± 2% discocytes. (b) C = 0.8 mM, AFM 3D-images 30 × 30 μm, 85 ± 5% deformed discocytes and stomatocytes. (c) C = 1.2 mM, AFM 3D-images 30 × 30 μm, 78 ± 6% planocytes and rising of topological “grain” defects. (d) C = 1.5 mM, AFM 3D-images 30 × 30 μm, 84 ± 10% cells with “grains”-containing domains on the membrane surface. (e) C = 2.5 mM, AFM 3D-images 30 × 30 μm, 79 ± 8% cells with merging domains and spheroechinocytes. Time of incubation was 60 min for (a–e). (f) The dependence of domains arising on incubation time ti for the same hemin concentration C = 1.5 mM (t1 = 3 min, t2 = 60 min and t3 = 180 min). Statistical processing performed by 100 × 100 μm AFM 3D-images of cells in monolayers in 45 smears. (60–100 cells in each image) for each concentration and each time.

Mentions: A representative AFM image (100 × 100 μm) of cells in the control smear is shown in Fig. 1a. Forty five images, each including 60–100 cells, were analyzed for each hemin concentration and each incubation time. Scanning of the fields 30 × 30 μm (Fig. 1 a–f) and 10 × 10 μm (Fig. 2, 3) was performed for more detailed AFM images.


Transformation of membrane nanosurface of red blood cells under hemin action.

Kozlova E, Chernysh A, Moroz V, Gudkova O, Sergunova V, Kuzovlev A - Sci Rep (2014)

Stages of transformation of RBC shape and its membrane under the hemin action for different concentrations C.(a) C = 0 (control), AFM 3D- images 100 × 100 μm and 30 × 30 μm of cells in control smear, 95 ± 2% discocytes. (b) C = 0.8 mM, AFM 3D-images 30 × 30 μm, 85 ± 5% deformed discocytes and stomatocytes. (c) C = 1.2 mM, AFM 3D-images 30 × 30 μm, 78 ± 6% planocytes and rising of topological “grain” defects. (d) C = 1.5 mM, AFM 3D-images 30 × 30 μm, 84 ± 10% cells with “grains”-containing domains on the membrane surface. (e) C = 2.5 mM, AFM 3D-images 30 × 30 μm, 79 ± 8% cells with merging domains and spheroechinocytes. Time of incubation was 60 min for (a–e). (f) The dependence of domains arising on incubation time ti for the same hemin concentration C = 1.5 mM (t1 = 3 min, t2 = 60 min and t3 = 180 min). Statistical processing performed by 100 × 100 μm AFM 3D-images of cells in monolayers in 45 smears. (60–100 cells in each image) for each concentration and each time.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4129419&req=5

f1: Stages of transformation of RBC shape and its membrane under the hemin action for different concentrations C.(a) C = 0 (control), AFM 3D- images 100 × 100 μm and 30 × 30 μm of cells in control smear, 95 ± 2% discocytes. (b) C = 0.8 mM, AFM 3D-images 30 × 30 μm, 85 ± 5% deformed discocytes and stomatocytes. (c) C = 1.2 mM, AFM 3D-images 30 × 30 μm, 78 ± 6% planocytes and rising of topological “grain” defects. (d) C = 1.5 mM, AFM 3D-images 30 × 30 μm, 84 ± 10% cells with “grains”-containing domains on the membrane surface. (e) C = 2.5 mM, AFM 3D-images 30 × 30 μm, 79 ± 8% cells with merging domains and spheroechinocytes. Time of incubation was 60 min for (a–e). (f) The dependence of domains arising on incubation time ti for the same hemin concentration C = 1.5 mM (t1 = 3 min, t2 = 60 min and t3 = 180 min). Statistical processing performed by 100 × 100 μm AFM 3D-images of cells in monolayers in 45 smears. (60–100 cells in each image) for each concentration and each time.
Mentions: A representative AFM image (100 × 100 μm) of cells in the control smear is shown in Fig. 1a. Forty five images, each including 60–100 cells, were analyzed for each hemin concentration and each incubation time. Scanning of the fields 30 × 30 μm (Fig. 1 a–f) and 10 × 10 μm (Fig. 2, 3) was performed for more detailed AFM images.

Bottom Line: The process of formation of "grains" was dependent on the hemin concentration and incubation time.The possible mechanism of membrane nanostructure alterations is proposed.This research can be used to study the cell intoxication and analyze the action of various agents on RBC membranes.

View Article: PubMed Central - PubMed

Affiliation: V.A. Negovsky Scientific Research Institute of General Reanimatology RAS, Moscow, Russian Federation.

ABSTRACT
Hemin is the product of hemoglobin oxidation. Some diseases may lead to a formation of hemin. The accumulation of hemin causes destruction of red blood cells (RBC) membranes. In this study the process of development of topological defects of RBC membranes within the size range from nanoscale to microscale levels is shown. The formation of the grain-like structures in the membrane ("grains") with typical sizes of 120-200 nm was experimentally shown. The process of formation of "grains" was dependent on the hemin concentration and incubation time. The possible mechanism of membrane nanostructure alterations is proposed. The kinetic equations of formation and transformation of small and medium topological defects were analyzed. This research can be used to study the cell intoxication and analyze the action of various agents on RBC membranes.

Show MeSH
Related in: MedlinePlus