Fig. 3.
Diabetes enhances integrin αIIbβ3 mediated platelet adhesion under shear. a Hirudinated whole blood from non-DM or DM mice were perfused through fibrinogen (FGN) matrices for 5 min at 600 s−1, and the number of adherent platelets determined at the indicated perfusion times. b Isolated mouse platelets were allowed to adhere to FGN matrices under static conditions for 5 min, and the number of adherent platelets quantitated. c Hirudinated whole blood from non-DM and DM mice were pretreated with amplification loop blockers (ALBs): indomethacin (10 μM), MRS2179 (100 μM), and 2-MeSAMP (10 μM), then perfused over spread platelets at 1800 s−1. The number of adherent platelets per spread platelet was quantified at 30 s intervals for 120 s. d Hirudinated mouse whole blood was perfused over mouse vWF matrices at 600, 1800, and 5000 s−1, as detailed under Methods section. The numbers of stationary (−integrilin) vs. transient/rolling (+integrilin) platelets were determined at 1 min perfusion time. e Hirudinated mouse whole blood was perfused through FGN matrices at 600 s−1 for 1 min, and adherent platelets were fixed immediately. Representative DIC images depict the discoid morphology of the adherent non-DM and DM mouse platelets. Scale bars = 5 μm. f Platelets isolated from non-DM and DM mice were reconstituted with isolated red blood cells (RBCs) from the same (autologous) or counterpart (heterologous) mice, prior to perfusion over matrices at 600 s−1. The number of adherent non-DM and DM platelets was determined at the indicated perfusion times. g Hirudinated whole blood from non-DM or DM humans was perfused through FGN matrices for 5 min at 300 s−1, and the number of adherent platelets determined at 5 min perfusion time. Results performed with human samples are expressed as mean ± s.e.m. of n = 16 for non-DM and n = 18 for DM patients. All other results represent n = 3 mice, with each perfusion performed in duplicates or triplicates. NS = not significant, p ≥ 0.05; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, assessed by unpaired, two-tailed Student’s t-test