Figure 4.

Cells with diabetes and dysfunctions in traction force and intercellular adhesion show disrupted asymmetric cell alignments. (a) The cell alignments and (b) histogram of the mean alignment angles of diabetic aortic ECs and ECs treated with blebbistatin (10 μM) and EDTA (50 μM) to inhibit actomyosin activity or intercellular adhesion, respectively, in micropatterned vascular sheets (D = 0.55) are shown. The angles of cell alignment within different micropatterns are illustrated as either positive (green) or negative (red) arrows in (a). Scale bar, 100 μm. Note the decreased asymmetric cell alignment in micropatterns with diabetic ECs and treatments with blebbistatin and EDTA. (c) Quantified traction force magnitude (red curves), asymmetric cell alignment (blue curves), cell-cell force magnitude (cyan curves), and mean alignment angle (purple curves) at different radial positions in micropatterns with diabetic ECs and different drug treatments are shown. Note the disturbed correlation between traction force and radial asymmetric cell alignments or cell-cell force and mean alignment angle after drug treatments. Error bars represent ± standard errors. (d) Quantified mean alignment angles in micropatterns with diabetic ECs and under different pharmacological treatments are shown. Note that inhibiting traction force or intercellular adhesion decreased cell alignment with larger mean alignment angles in the inner and outer regions of the ring patterns, whereas there was no significant difference observed in the middle regions. P-values were calculated using one-way ANOVA. *, P < 0.05. (e) A table shows the quantified mechanical force magnitude and asymmetric cell alignment in micropatterns with diabetic ECs (n = 11) and under different pharmacological treatments (n ≥ 20). Note the decreased traction force and asymmetric cell alignment in diabetic ECs and micropatterns treated with blebbistatin.