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. Author manuscript; available in PMC: 2018 Jun 23.
Published in final edited form as: Circ Res. 2017 Apr 20;121(1):31–42. doi: 10.1161/CIRCRESAHA.117.310694

Figure 2. Creation of intimal lesions for experimental probing of mechanisms related to superficial erosion in mice.

Figure 2

(A) Experimental protocol showing carotid electric injury (yellow segment) performed at day 0 and followed in 4 weeks by completed endothelial reconstitution. Composition of the neointima in post-injured arteries in ECs (CD31), vascular smooth muscle cells (SMA), macrophages (Mac3), and hyaluronan (HA). The insets show higher magnification views for each representative image. Carotid lumen (star) and internal elastic laminae (arrow) are shown. ni: neointima. The arrow represents flow orientation. LCCA: left common carotid injury. QPCR analysis was performed on normal vs previously injured carotid for the expression of extracellular matrix related mRNAs (B). Data are expressed in 2−ΔCt to β-actin. Each dot represents data from one animal. Mann–Whitney U test. (C) Western blot and relative quantification showing overexpression of E-Selectin and VCAM-1 in normal vs previously-injured arteries, n=4. Quantitative PCR on lysates from human saphenous vein ECs cultured 48h on native gelatin coating (untreated) vs coating enriched with hyaluronan (HA) of various molecular weights (D) or vs 100 or 500μg/ml of 5kDa HA-enriched coating (E). Data are expressed in 2−ΔΔCt to untreated condition, mean ± s.e.m, n=6 per group, *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001. Paired t-test (F,G,H) Placement of a cone-shape polyethylene cuff around the LCCA in mice and secured by a circumferential suture. Effect of non-constrictive control cuff (g) or constrictive cuff placement on flow dynamics in carotid arteries (H). Doppler-coupled ultrasonography showing velocity patterns along the downstream flow (left, arrow) and the velocity peak downstream of the cuff (right, arrow) measured at 1.5 mm downstream of the cuff (arrowhead). Computational fluid dynamics using finite volume analysis verified the extent of flow perturbation and recirculation. Longitudinal cross-sectional simulation of the effect of a non-constrictive cuff (G) or a constrictive cuff (H) showing a normalized axial velocity contour with superimposed velocity vectors (G,H). LCCA: left common carotid artery, LCCA: left common carotid artery, ECA: external carotid artery, ICA: internal carotid artery, NC: non-constrictive cuff, CC: constrictive cuff.