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. 2024 Jul 11;44(9):2118–2135. doi: 10.1161/ATVBAHA.124.321000

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© 2024 The Authors.

Arteriosclerosis, Thrombosis, and Vascular Biology is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited.

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Figure 2. — Fast-progressive aortic stenosis (FP-AS) and slow-progressive aortic stenosis (SP-AS) show typical valvular expression patterns of valve infiltrating inflammatory cells, platelets, and macrophage MIF (macrophage migration inhibitory factor). A, Representative immunohistological stainings of SP-AS (n=110) and FP-AS (n=108) to calculate the total aortic valve cell count (n), CD68⁺ macrophages, CD16⁺ monocytes, CD14⁺ monocytes, CD42b⁺ platelets, and MIF⁺ staining in matching areas of consecutive aortic valve (AV) tissue sections. Plotted: median±interquartile range (IQR); statistics: Mann-Whitney U test. B, Representative immunohistological stainings comparing aortal and ventricular aortic valve side in SP-AS (n=11) and FP-AS (n=14) to analyze the cell count (n) and CD42b⁺ positive areas in the aortal and the ventricular side of the AV. Plotted: median±IQR; statistics: Kruskal-Wallis test. C, In vitro analysis of cell culture of valvular interstitial cells (VICs) in control, proinflammatory, and pro-osteogenic medium. Vimentin and αSMA expression on VICs in control, proinflammatory, and pro-osteogenic medium without stimulation, under stimulation with MIF, coincubation with MIF and its antagonist ibudilast, and under inhibition by ibudilast. Plotted: mean±SD, n=4; statistics: 1-way ANOVA. Tukey multiple comparison test (P<0.05) was used to correct for multiple comparisons. D, ALP (alkaline phosphatase) expression on VICs in control, proinflammatory, and pro-osteogenic medium without stimulation, understimulation with MIF, coincubation with MIF and its antagonist ibudilast, and under inhibition by ibudilast. Proinflammatory and pro-osteogenic medium mimic an inflammatory (FP-AS) or calcifying (SP-AS) valvular and systemic phenotype as described before. Plotted: mean±SD, n=4; statistics: 1-way ANOVA. Tukey multiple comparison test (P<0.05) was used to correct for multiple comparisons. HE indicates hematoxylin and eosin.