ABSTRACT
During diabetes, the characteristic hyperglycemia can induce red blood cell glycation. Several researchers have proposed different protocols to perform an in vitro model to study this phenomenon. In this article, some of the most important in vitro glycation protocols available in the bibliography were compared to each other. The incubation parameters as the suspension medium, glucose concentration, red blood cell concentration, time, and temperature were analyzed. Also, several assays were carried out in our laboratory, and glycated hemoglobin, erythrocyte aggregation and viscoelasticity were determined for the protocol validation. Based on the bibliographic analysis and our experimental results, an optimal protocol for in vitro glycation of red blood cells is presented.
Subject(s)
Blood Chemical Analysis/methods , Erythrocytes/metabolism , Adult , Biochemical Phenomena , Blood Glucose/metabolism , Blood Viscosity , Elasticity , Erythrocyte Aggregation , Erythrocytes/drug effects , Glucose/pharmacology , Glycated Hemoglobin/chemistry , Glycated Hemoglobin/metabolism , Glycosylation , Hematocrit , Humans , In Vitro Techniques , Male , Models, BiologicalABSTRACT
Several studies report flow disturbance and microcirculation disorders upon anesthesia treatment. These alterations are often related to blood rheology changes. In this work, it was attempted to make a detailed description of the alterations in erythrocyte mechanical properties by the action of propofol, remifentanil, and vecuronium. For this, an in vitro study was performed on red blood cell samples from healthy donors incubated with solutions of propofol (4 µg/mL whole blood), remifentanil (10 ng/mL plasma), and vecuronium (0.15 µg/mL plasma). Erythrocyte viscoelastic parameters were determined by octuplicate using a Reómetro Eritrocitario. Also, a Wilcoxon signed rank-test with Yates correction for continuity was performed to analyze the overall alteration in the mechanical properties of erythrocytes. Statistical analysis showed that the three studied anesthetics changed the erythrocyte mechanical properties at different parts of the membrane. These results would imply an interaction of these anesthetics with the erythrocyte membrane. Finally, this could conduce to alterations in microcirculation.
