Figure 1.
Proliferation (A, B) and migration (C, D) of proliferative (A, C) and quiescent (B, D) SMCs in response to treatment with DS-SILY20. Proliferation and migration of proliferative SMCs significantly decreased with increased concentrations of DS-SILY20. Proliferation in quiescent SMC cultures was not altered with addition of DS-SILY20 at any concentration; however, quiescent SMC migration decreased with the addition of 1 and 10 μM DS-SILY20. * represents significance from control non-treated cells. (N>6).
Figure 2.
Protein expression of (A) proliferative and (B) quiescent SMCs in response to treatment with DS-SILY20. A significant decrease in protein expression was observed in proliferative SMC cultures treated with 10 μM DS-SILY20, while no changes in protein synthesis was exhibited in proliferative SMC cultures treated with lower concentrations of DS-SILY20 or quiescent cultures with the addition of any concentration DS-SILY20. * represents significance from control non-treated cells. (N>6).
Figure 3.
Cytokine produced (A, C, E, G) proliferative and (B, D, F, H) quiescent SMCs in response to DS-SILY20. The amount of (A, B) IFN-γ, (C, D) IL-1β, (E, F) IL-6, and (G, H) TNF-α produced by cultured SMCs was measured 24 hrs post-treatment. For proliferative SMC cultures, a general trend was observed such that as the concentration of DS-SILY20 increased, cytokine production decreased. Significant reductions in IFN-γand TNF-α were also observed in quiescent SMC cultures with the addition of DS-SILY20. * represents significance from control non-treated cells. (Nγ6).
Figure 4.
Thrombomodulin expression from (A) proliferative and (B) quiescent SMCs in response to treatment with DS-SILY20. Thrombomodulin expression decreased with the addition low concentrations of DS-SILY20; however, the addition of either 1 or 10 μM DS-SILY20 resulted in significantly increased thrombomodulin production. Thrombomodulin production was not altered with the addition of DS-SILY20 in quiescent SMC cultures. * represents significant increase from control non-treated cells; # represents significant decrease from control non-treated cells. (N>4).
Figure 5.
Platelet Deposition and Activation.
SEM images of acute platelet response on artery wall in vessels treated with (A, C) DS-SILY20 and (B, D) sham saline control. Saline-treated arteries exhibited significant platelet coverage with numerous projections extending from activated platelets; platelets were scarcely visible on denuded arteries treated with DS-SILY20. Arrowheads indicate areas of platelet coverage. Magnification = 3,500x (A, B); 35,000x (C, D). Scale bar = 20 μm (A, B); 2 μm (C, D).
Figure 6.
Representative histology sections of (A, B) arteries with stents and (C, D) arteries without stents treated with (A, C) saline or (B, D) 10 μM DS-SILY20. Arrowheads indicate location of some stent struts; internal elastic lamina (dotted line) and lumenal border (solid line) are identified, indicating the boundaries of the neointima formed following injury. (E) Neointimal hyperplasia was quantified by measuring the distance from a stent strut or the elastic lamina to the vessel lumen in arteries with or without stents, respectively. A significant reduction in neointimal hyperplasia with DS-SILY20 treatment was observed, compared with saline sham controls, for both stented and non-stented artery segments. Six measurements were taken for each artery. Analysis with stents: sham (n=4), DS-SILY20 (n=3); without stents: sham (n=8), DS-SILY20 (n=5). Scale bar = 1 mm.* represents significance from sham-treated vessels.