Figure 3.

Shock‐wave therapy: effects on endothelial cells and angiogenesis ex vivo. A, Activation of Akt and ERK after SWT. The MAPK/ERK and Akt kinases play a crucial role in the regulation of angiogenesis. SWT induces phosphorylation of Akt after 30 min, whereas ERK is activated after 30 min, 2 h, 4 h, and 6 h. All experiments were performed at least in triplicate. B, SWT promotes endothelial cell proliferation and survival. HUVECs were treated with SWT, and DAPI‐stained nuclei were quantified 24 h after treatment. We found increased numbers of endothelial cells after SWT compared with untreated controls. To analyze effects on cell survival, HUVECs were treated with SWT after starvation, whereas controls remained untreated. Apoptotic cells were quantified using a TUNEL assay. SW‐treated cells showed improved survival compared with untreated controls. (scale bar=100 μm) (***P<0.001 vs CTR, ****P<0.0001 vs CTR). All experiments were performed at least in triplicate. C, Induction of capillary sprouting in aortic rings. Murine aortas were cultured in matrigel and treated with SWT. Capillary sprouts were quantified 7 d after treatment. SWT promoted capillary sprouting compared with untreated controls. Treatment of aortic rings with supernatant from SW‐treated cells improved capillary sprouting as well; however, not significantly. (n=6 per group, ****P<0.0001 vs CTR). CTR indicates control; DAPI, 4′,diamidino‐2‐phenylindole; ERK, extracellular‐signal regulated kinase; HUVEC, human umbilical vein endothelial cell; MAPK, mitogen‐activated protein kinase; SN, supernatant; SWT, shock wave therapy; TUNEL, terminal deoxynucleotidyl transferase dUTP nick‐end labeling.