Prolonged White Matter Inflammation After Cardiopulmonary Bypass and Circulatory Arrest in a Juvenile Porcine Model Ludmila Korotcova, MD, Sonali Kumar,

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Prolonged White Matter Inflammation After Cardiopulmonary Bypass and Circulatory Arrest in a Juvenile Porcine Model Ludmila Korotcova, MD, Sonali Kumar, BS, Kota Agematsu, MD, Paul D. Morton, PhD, Richard A. Jonas, MD, Nobuyuki Ishibashi, MD The Annals of Thoracic Surgery Volume 100, Issue 3, Pages 1030-1037 (September 2015) DOI: 10.1016/j.athoracsur.2015.04.017 Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 (A) Study design. (B) Subdivision of cerebral white matter (WM). (C) Iba1+ microglia cell and the counting frame for analysis. Microglia is counted if it lies entirely within the counting frame or if it touches an acceptance line without touching a rejection line. (CA = circulatory arrest; CPB = cardiopulmonary bypass; FF = full-flow perfusion; I/R = ischemia-reperfusion and reoxygenation; NT = normothermia; PVWM = peri-ventricular white matter; RW = rewarming; SIRS = systemic inflammatory response syndrome.) The Annals of Thoracic Surgery 2015 100, 1030-1037DOI: (10.1016/j.athoracsur.2015.04.017) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Cardiopulmonary bypass induced cerebral ischemia-reperfusion and reoxygenation result in acute microglia activation. A, B, Iba1+ microglia in control (A) and after severe cardiopulmonary bypass (CPB) (B). (C) Iba1+ microglia cell number in 7 white matter (WM) regions on post-CPB day 3. (D) Iba1+ microglia cell number in 3 fiber categories on post-CPB day 3. (* = p < 0.001 versus control and mild by one-way analysis of variance [ANOVA] with Bonferroni comparisons; ** = p < 0.001 versus control and mild by two-way ANOVA with Bonferroni comparisons. Scale bar = 50 μm. Data are shown as mean ± standard error of the mean; (C) and (D) n = 5–6 each). (CA = circulatory arrest; FF = full-flow perfusion; PVWM = peri-ventricular white matter; RW = rewarming.) The Annals of Thoracic Surgery 2015 100, 1030-1037DOI: (10.1016/j.athoracsur.2015.04.017) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Prolonged microglia activation after cardiopulmonary bypass is area dependent. (A) Iba1+ microglia cell number in 7 white matter (WM) regions on post-cardiopulmonary bypass (CPB) week 4. (B) and (C) Iba1+ microglia cell number in 3 fiber categories on post-CPB week 4. (D) Ki67+ proliferating cell number in 3 fiber categories on post-CPB week 4. (* p = 0.01 vs control; ** p < 0.01 vs control by one-way analysis of variance with Bonferroni comparisons. Data are shown as mean ± standard of the mean [n = 5–6 each]). (PVWM = peri-ventricular white matter.) The Annals of Thoracic Surgery 2015 100, 1030-1037DOI: (10.1016/j.athoracsur.2015.04.017) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Microglia reaction to cardiopulmonary bypass is area dependent and varies according to brain insults. (A) Mean Iba1+ microglia cell number on post-CPB day 3 and week 4. (B) Change of Iba1+ microglia cell number after CPB in 3 fiber categories. Change of Iba1+ microglia cell number in (C) commissural, (D) projection, and (E) association fibers. (∗ p = 0.05 vs control; ∗∗ p < 0.01 vs control; ∗∗∗ p < 0.001 vs control and mild by one-way analysis of variance (ANOVA) with Bonferroni comparisons. CPB = cardiopulmonary bypass; †, p = 0.05; ‡, p < 0.001 vs control and mild by two-way ANOVA with Bonferroni comparisons. Data are shown as mean ± standard error of the mean (n = 5–6 each). (F = F-statistics in the ANOVA.) The Annals of Thoracic Surgery 2015 100, 1030-1037DOI: (10.1016/j.athoracsur.2015.04.017) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions