Abstract

The mechanical properties and deformability of Red Blood Cells (RBCs) are important determinants of blood rheology and microvascular hemodynamics. The objective of this study is to quantify the mechanical properties and wall shear stress experienced by the RBC membrane during capillary plug flow in vivo utilizing high speed video recording from intravital microscopy, biomechanical modeling, and computational methods. Capillaries were imaged in the rat cremaster muscle pre- and post-RBC transfusion of stored RBCs for 2-weeks. RBC membrane contours were extracted utilizing image processing and parametrized. RBC parameterizations were used to determine updated deformation gradient and Lagrangian Green strain tensors for each point along the parametrization and for each frame during plug flow. The updated Lagrangian Green strain and Displacement Gradient tensors were numerically fit to the Navier-Lame equations along the parameterized boundary to determined Lame's constants. Mechanical properties and wall shear stress were determined before and transfusion, were grouped in three populations of erythrocytes: native cells (NC) or circulating cells before transfusion, and two distinct population of cells after transfusion with stored cells (SC1 and SC2). The distinction, between the heterogeneous populations of cells present after the transfusion, SC1 and SC2, was obtained through principle component analysis (PCA) of the mechanical properties along the membrane. Cells with the first two principle components within 3 standard deviations of the mean, were labeled as SC1, and those with the first two principle components greater than 3 standard deviations from the mean were labeled as SC2. The calculated shear modulus average was 1.1±0.2, 0.90±0.15, and 12 ± 8 MPa for NC, SC1, and SC2, respectively. The calculated young's modulus average was 3.3±0.6, 2.6±0.4, and 32±20 MPa for NC, SC1, and SC2, respectively. o our knowledge, the methods presented here are the first estimation of the erythrocyte mechanical properties and shear stress in vivo during capillary plug flow. In summary, the methods introduced in this study may provide a new avenue of investigation of erythrocyte mechanics in the context of hematologic conditions that adversely affect erythrocyte mechanical properties.

Highlights

  • Red Blood Cells (RBCs) are highly deformable biconcave particles with diameter 8 μm that constitute ∼45% of total blood volume

  • The objective of this study is to quantify the mechanical properties and wall shear stress experienced by RBC membranes during capillary plug flow in vivo utilizing intravital imaging and computational methods

  • In the population of erythrocytes analyzed from animals transfused with stored cells, RBCs with both principle components greater than 3 standard deviations from the mean principle component of native RBCs were identified and analyzed as a separate population for all calculations (Figure 5B). 68.6% (24/35) of RBCs analyzed from animals subjected to exchange transfusion were found to have both components within 3 standard deviations of the native RBC principle components (SC1)

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Summary

Introduction

Red Blood Cells (RBCs) are highly deformable biconcave particles with diameter 8 μm that constitute ∼45% of total blood volume. The non-linear viscoelastic mechanical properties of the RBC membrane results in context-dependent elastodynamic behavior that stems from changes in shear modulus (Stokke et al, 1986; Secomb and Hsu, 1995). While tank-treadmilling motion increases membrane energy dissipation, cytoplasmic energy dissipation is minimized, preventing hemolysis and capillary plugging (Lanotte et al, 2016). Together, these aspects of RBCs impart the ability for these cells to withstand large shear stresses in the larger vessels while being able to deform through capillaries with ease

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