Fig. 8

CAM depletion reduces IA formation and ameliorates severity of IA ruptures. a Schematic representation of the experimental design. b Representative flow cytometry dot-plots and gating strategy used for quantification of CD206⁺ macrophages and microglia, with vehicle or clodronate-liposomes. c CLO-liposomes significantly reduce the number of CD206⁺ macrophages even 10 days post-depletion, whereas microglia number stayed unaltered. Flow cytometry quantification of CD206⁺ macrophages and Iba1⁺ microglia (d), treated either with vehicle or CLO-liposomes (N = 4, *P < 0.05, Mann–Whitney U test). e Average number of IAs developed per animal. Immunofluorescence analysis data (on the left) show that CLO-treated mice develop significantly fewer IAs compared to the control group (*P < 0.05, N = 6 CLO-treated mice/N = 4 PBS-treated mice, Mann–Whitney U test). Longitudinal MR studies over the timeline of 15 days confirmed the existing difference (on the right). f CLO-treated mice have decreased global rupture rate (*P < 0.05, n = 14 PBS-treated mice/n = 13 CLO-treated mice; PBS versus CLO: 72% versus 43%, Fisher exact test). g Aneurysm repartition between the PBS and CLO-treated mice during the period of 15 days (assessed on MR scans). h CAM depletion significantly reduces the hemorrhage volumes after the IA rupture, assessed by the MRI scans from T2*-w sequences (*P < 0.01; Mann–Whitney U test). i, j Wall-to-lumen ratio and intima-media thickness comparison between the group revealed no significant difference in dimensions of IAs (**P < 0.01, ***P < 0.001, N = 4 CLO-treated mice, n = 7 IAs/n = 6 contra arteries; N = 4 PBS-treated mice, n = 16 IAs/n = 8 contra arteries. Two-way ANOVA). Statistical analysis was performed using two-way analysis of variance (ANOVA) with post hoc Dunnett adjustment; *P < 0.05, **P < 0.01, ***P < 0.001. Further detailed information on statistics is provided in the Methods & Materials