Glucose Concentration Affects Fibrin Clot Structure and Morphology as Evidenced by Fluorescence Imaging and Molecular Simulations

Abstract

Although in vivo studies have been conducted in the past to determine hyperglycemic effects and influence on clotting risk in patients with diabetes, the true extent of hyperglycemia on unstable and spontaneous clot formation remains highly debated. Factors such as increased glycation, elevated fibrinogen concentration, elevated prothrombin levels, and decreased plasminogen are known to influence fibrin conversion, clot morphology, and thrombus formation in these individuals. In this regard, the isolated effects of hyperglycemia on irregular fibrin clot formation were investigated in a controlled fibrinogen system. In this study, fibrin clot characteristic differences at 3 glucose concentrations were analyzed to determine the effects of glucose concentration on fibrinogen glycation and fibrin clot morphology using confocal microscopy, glycation quantification, molecular simulations, and image processing methods. Algorithms coupled with statistical analysis support in vivo findings that hyperglycemia increases fibrinogen glycation, with ensuing altered fibrin clot structure characteristics. Our experimental and molecular simulation results consistently show an increased glucose adsorption by fibrinogen with increased glucose concentration. Significant differences in clot structure characteristics were observed, and the results of this work can be used to further develop diagnostic tools for evaluating clotting risk in individuals with hypercoagulable and hyperglycemic conditions.

Fibrinogen Glycation and Presence of Glucose Impair Fibrin Polymerization—An In Vitro Study of Isolated Fibrinogen and Plasma from Patients with Diabetes Mellitus

“…Additionally, only a slight insignificant reduction of V max was observed for the glucose-incubated fibrinogen when glucose was absent during polymerization, compared to control fibrinogen not subjected to the influence of glucose. Hood et al have reported a reduced fiber overlap in a fibrin subjected to glucose, decreased fibrin length and fractal dimension, and increased porosity of fibrin incubated in media containing 10 mM glucose compared to glucose-free control media [ 18 ]. Native fibrinogen was incubated with glucose (6 or 10 mM) for 48 h, following which, a fibrin clot was polymerized in the presence of glucose.…”

“…The extent of glucose incorporation into fibrinogen, determined as the difference between glucose concentrations recorded for time 0 and time 48 h, was found to be the highest under the hyperglycemic condition. Molecular docking and molecular dynamics simulation suggest that glucose adsorption on fibrinogen is highly dependent on the glucose concentration and characterized by a high affinity of glucose molecule to the hydrophobic amino residues [ 18 ]. In addition, Lippi et al have note that fibrinogen concentration measured by the Clauss method significantly decreased with increasing glucose contamination (0–5–10–20%) [ 19 ].…”

“…The presence of glycated fibrinogen was confirmed in both the plasma from diabetes patients [ 7 , 11 , 27 ], and in the samples incubated in vitro with glucose [ 18 , 28 ]. Generally, several methods have been used to determine fibrinogen glycation, such as the hydroxymethylfurfural assay [ 27 ], Gly-Pro ® assay [ 7 ] or NBT assay [ 11 ].…”

Углеводная Модификация Фибриногена И Фибрина В Условиях Гипергликемии Как Фактор Изменения Структуры И Свойств Тромба (Обзор Литературы)

“…Строение и свойства тромба являются важными детерминантами болезней и смертности, связанных с сердечно-сосудистой патологией [1][2][3][4][5][6][7]. Одним из основных факторов, определяющих характеристики тромба, являются изменения в молекуле фибриногена, которые могут влиять на скорость гемокоагуляции, структуру фибринового матрикса и его восприимчивость к фибринолизу [1,[8][9].…”

Practical Implementation of Molecular Dynamics Code for Beginners Using Python

“…However, often there arises a problem which needs customized molecular simulations. This can be due to the boundary condition, the construction of the system itself, approximate models for introducing new force fields (12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30) or can be coupling between different time or length scales. To account these studies, graduate students often write their own code which is a cumbersome task (20,21,28).…”