Membrane Lesions and Reduced Life Span of Red Blood Cells in Preeclampsia as Evidenced by Atomic Force Microscopy

Abstract

Preeclampsia (PE) presents with maternal de novo hypertension and significant proteinuria and is one of the leading causes of maternal and perinatal morbidity and mortality with unknown etiology. The disease is associated with inflammatory vascular response and severe red blood cell (RBC) morphology changes. This study examined the nanoscopic morphological changes of RBCs from PE women versus normotensive healthy pregnant controls (PCs) and non-pregnant controls (NPCs) applying atomic force microscopy (AFM) imaging. The results revealed that the membrane of fresh PE RBCs differed significantly from healthy ones by the presence of invaginations and protrusions and an increased roughness value (Rrms) (4.7 ± 0.8 nm for PE vs. 3.8 ± 0.5 nm and 2.9 ± 0.4 nm for PCs and NPCs, respectively). PE-cells aging resulted in more pronounced protrusions and concavities, with exponentially increasing Rrms values, in contrast to the controls, where the Rrms parameter decreased linearly with time. The Rrms, evaluated on a 2 × 2 µm2 scanned area, for senescent PE cells (13 ± 2.0 nm) was significantly higher (p < 0.01) than that of PCs (1.5 ± 0.2 nm) and NPCs (1.9 ± 0.2 nm). Furthermore, the RBCs from PE patients appeared fragile, and often only ghosts were observed instead of intact cells at 20–30 days of aging. Oxidative-stress simulation on healthy cells led to RBC membrane features similar to those observed for PE cells. The results demonstrate that the most pronounced effects on RBCs in PE patients are related to impaired membrane homogeneity and strongly altered roughness values, as well as to vesiculation and ghost formation in the course of cell aging.

Red Blood Cell Storage with Xenon: Safe or Disruption?

“…As reported in our previous publications and other studies [ 27 , 45 , 46 , 47 , 48 ], the observed changes in cell shape may be associated with damage to local membrane and cytoskeletal connections. The cytoskeleton comprises a network of proteins, predominantly spectrin tetramers, linked with complexes of actin, protein 4.1R, ankyrin, as well as tropomyosin, tropomodulin, adducin, and dematin [ 48 , 49 ].…”

“…The cytoskeleton comprises a network of proteins, predominantly spectrin tetramers, linked with complexes of actin, protein 4.1R, ankyrin, as well as tropomyosin, tropomodulin, adducin, and dematin [ 48 , 49 ]. The integrity and flexibility of the membrane is largely maintained by the interaction of the spectrin-ankyrin band 3 complex [ 47 , 50 ]. In addition, disruption of these bonds can cause spectrin tetramers to dissociate into dimers [ 51 ], and the protein structures of the matrix network can undergo polymerization and aggregation [ 52 ].…”

“…In addition, a 2.5-fold increase in membrane stiffness was observed. The aggregation of band 3 protein results in decreased affinity for the cytoskeletal complex, which in turn increases membrane stiffness and reduces cellular functionality [ 47 , 50 , 53 ]. Erythrocyte stiffness is an important blood flow parameter because stiffer erythrocytes have more difficulty passing through small capillaries to deliver oxygen.…”

Assessment of Red Blood Cell Aggregation in Preeclampsia by Microfluidic Image Flow Analysis—Impact of Oxidative Stress on Disease Severity

“…The extensive decrease of RBC membrane elasticity or de-formability in some pathological conditions is indicative of abnormal cell morphology and damaged RBC capacity to adequately adapt its shape to navigate blood vessels, resulting in impaired hemorheology [58]. Recently, we have reported impaired membrane integrity and the presence of RBCs with unusual morphology in preeclamptic women [59]. A great percentage of schistocytes (fragmented red blood cells) was also found in peripheral blood smears of the samples from patients with hypertensive disorders of pregnancy [60].…”

“…Oxidative stress was chemically induced using hydrogen peroxide (H1009 Sigma-Aldrich Pty Ltd., an affiliate of Merck KGaA, Darmstadt, Germany) on newly isolated RBCs from three healthy pregnant women who were not included in the control group PC according to the protocol described in [59]. In brief, RBCs were treated for 4 h at 37 • C with 200 mM, 300 mM, and 400 mM H 2 O 2 .…”