Glycation of Fibrinogen and its Effect on Fibrin Clot Formation and Structure
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Tutor / directorPerez-Luna, Victor H.
Document typeBachelor thesis
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This research project will test the hypothesis that glycat ion of fibrinogen can affect the propert ies of the fibrin clot , its formation, and degradat ion. Studying those changes in vitro could provide a molecular explanation for some of the observed complications of diabetes such as vascular disease and impai red wound healing. The first part of the research project includes the study of the kinetics of fibrinogen glycation reaction using fluorescence and optical absorption spectroscopy. The extent of reaction will be quantified by determining the number of amine groups per fibrinogen molecule before and after glycation. In the second part of the project fibrinogen with different extents of glycation will be reacted with thrombin in order to form fibrin clots. The kinetics of fibrin clot formation will be studied by measuring the turbidity of the sample as the fibrin clot forms. The effect of glycation on fibrin clot formation will be determined from differences in the kinetics of this process. In addition to this, the structure of the fibrin clot will be examined using microscopy in order to observe potential differences in fiber´s structure. Enzymatic degradation of fibrin clots prepared with glycated fibrinogen will also be determined in order to fully understand the effects of glycation on fibrinogen. Using absorption spectroscopy and fluorescence emission is possible to confirm the formation of Amadori complex, and therefore, prove that the glycation reaction of fibrinogen occurred. Results were not as the expected results due to fibrinogen precipitation and/or experimental errors that are uncontrollable. Even so, the conclusion was that fibrin clots formed with plasma from diabetic patients or non-diabetic patients differ. Thus, fibrin clots from diabetic patients are denser and less porous compared with fibrin clots formed from non-diabetic plasma. So, an increase of glucose concentration in plasma implies a significant increase in fiber density and branch points. Also, the concentration of glucose implies a resistance in its fibrinolysis. However, despite of all the obtained results by many researchers, further investigation is necessary in this field.
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