Morphometric and Nanomechanical Features of Platelets from Women with Early Pregnancy Loss Provide New Evidence of the Impact of Inherited Thrombophilia

Abstract

Pregnancy is associated with hypercoagulation states and increased thrombotic risk, especially in women with thrombophilia. We combine atomic force microscopy (AFM) and flow cytometry to examine the morphology and nanomechanics of platelets derived from women with early pregnancy loss (EPL) and control pregnant (CP) and non-pregnant (CNP) women. Both control groups exhibit similar morphometric parameters (height and surface roughness) and membrane stiffness of platelets. EPL patients’ platelets, on the other hand, are more activated than the control groups, with prominent cytoskeletal rearrangement. In particular, reduced membrane roughness (22.9 ± 6 nm vs. 39.1 ± 8 nm) (p < 0.05) and height (692 ± 128 nm vs. 1090 ± 131 nm) (p < 0.05), strong alteration in the membrane Young modulus, increased production of platelets’ microparticles, and higher expression of procoagulant surface markers, as well as increased occurrence of thrombophilia (FVL, FII20210A, PLA1/A2, MTHFR C677T or 4G/5G PAI-1) polymorphisms were found. We suggest that the carriage of thrombophilic mutations triggers structural and nanomechanical abnormalities in platelets, resulting in their increased activation. The activation state of platelets can be well characterized by AFM, and the morphometric and nanomechanical characteristics might serve as a new criterion for evaluation of the cause of miscarriage and offer the prospect of an innovative approach serving for diagnostic purposes.

Mean platelet volume as an effective biomarker for predicting high-risk patients of hereditary thrombophilia: A retrospective study

“…The most commonly detected hereditary thrombophilias are usually related with genes coding for proteins in the coagulation cascade [ 20 – 24 ]. In hypercoagulated states due to hereditary thrombophilia, such as prothrombotic allele of PIA2, FVL, FII20210A, 677MTHFR, PLA1/A2, and 4G/4GPAI-1, a significant platelet activation was detected secondary to the changes in the surface roughness and morphology of the platelets [ 25 , 26 ]. The close association was observed between higher MPV values and recurrent pregnancy loss in patients with either homozygous FVL or PT mutations or was double heterozygotes for FVL and PT mutations [ 17 , 18 ].…”

Morphometric and Nanomechanical Screening of Peripheral Blood Cells with Atomic Force Microscopy for Label-Free Assessment of Alzheimer’s Disease, Parkinson’s Disease, and Amyotrophic Lateral Sclerosis

“…The membrane roughness of RBCs was drastically affected and that of PLTs to a much smaller extent, demonstrating lower values in the studied NDDs than in healthy cells (Figure 2C). The decrease in Rrms, accompanying the membrane smoothening, was much more Along with the cell shape, spreading area, and volume, membrane roughness and stiffness were also found useful for differentiation between healthy persons and patients with acute myocardial infarction [255], type 2 diabetes mellitus (emorheological and atomic force microscopy studies on the experimental clot formations in patients with type 2 diabetes mellitus), hypertension [256], transient ischemic attacks [257], and inherited thrombophilia [258]. We also reported that the PLT membranes in NDDs were significantly stiffer than the control PLTs (Figure 2D, [254]).…”

Hematological and Hemorheological Parameters of Blood Platelets as Biomarkers in Diabetes Mellitus Type 2. A Comprehensive Review

“…During PLTs activation, changes in the shape of PLTs appear first, from discoid to spherical, with actin-rich philopodia (cytoplasmic extrusions) interacting, flattening and spreading on the surfaces in the last of the activation phases [9][10][11]. The same authors characterized the PLTs activation stages morphometrically and nanomechanically by atomic force microscope (AFM) for diagnostic purposes.…”