Nanotechnology Approaches to Targeting Inflammation and Excitotoxicity in a Canine Model of Hypothermic Circulatory Arrest–Induced Brain Injury Joshua.

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Nanotechnology Approaches to Targeting Inflammation and Excitotoxicity in a Canine Model of Hypothermic Circulatory Arrest–Induced Brain Injury Joshua C. Grimm, MD, J. Trent Magruder, MD, Mary A. Wilson, PhD, Mary E. Blue, PhD, Todd C. Crawford, MD, Juan C. Troncoso, MD, Fan Zhang, PhD, Sujatha Kannan, PhD, Christopher M. Sciortino, MD, PhD, Michael V. Johnston, MD, Rangaramanujam M. Kannan, PhD, William A. Baumgartner, MD The Annals of Thoracic Surgery Volume 102, Issue 3, Pages 743-750 (September 2016) DOI: 10.1016/j.athoracsur.2016.02.077 Copyright © 2016 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Correlation of the total histopathology score with the neurobehavioral score after 1.5 hours of hypothermic circulatory arrest (HCA). For comparison with our previous published study [22], data include correlations for the 90-minute HCA animals and those for animals receiving cardiopulmonary bypass (CPB), 1 hour of HCA, or 2 hours of HCA. The Annals of Thoracic Surgery 2016 102, 743-750DOI: (10.1016/j.athoracsur.2016.02.077) Copyright © 2016 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Neuronal injury in the hippocampus after 1.5 hours of hypothermic circulatory arrest (HCA). (A and D) In the dentate gyrus of the dorsal hippocampus, neurons with apoptotic injury are shrunken and show clumped chromatin (arrows in A1, D1) and (B and E) are labeled with activated caspase 3 antibodies. (A–C) The animal with the highest neurobehavioral score had many more apoptotic neurons and increased activated caspase 3 expression, than (D–F) animals with less severe neurobehavioral scores. (C) Likewise, microglia show a more activated appearance in the more injured animal. (H&E = hematoxylin and eosin.) Large boxes (A-F) scale 50 μm. Small insets (A1 and D1) scale 25 μm. The Annals of Thoracic Surgery 2016 102, 743-750DOI: (10.1016/j.athoracsur.2016.02.077) Copyright © 2016 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Neuronal uptake of dendrimer (D)-cyanine 5 (Cy5) in the hippocampus after 1.5 hours of hypothermic circulatory arrest (HCA). (A) In the CA1 region of the dorsal hippocampus, neurons (neuronal class III β-tubulin [Tuj1]-labeled, green) with abnormal morphology (filled arrows) (B–C) contain D-Cy5 (red). (A–C) A neuron with more normal morphology (open arrow) contains very little D-Cy5. The ventral hippocampus typically exhibits less injury than the dorsal hippocampus after HCA [22]. (D–F) In the CA1 region of ventral hippocampus, there is some uptake of D-Cy5 in neuronal processes, but the somas have normal morphology and do not exhibit notable accumulation of D-Cy5. (Scale bar = 20 μm.) The Annals of Thoracic Surgery 2016 102, 743-750DOI: (10.1016/j.athoracsur.2016.02.077) Copyright © 2016 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Microglia and astrocyte uptake of dendrimer (D)-cyanine 5 (Cy5) in the hippocampus after 1.5 hours of hypothermic circulatory arrest (HCA). (A–C) Arrowheads indicate a microglial cell (Iba-1 labeled, green) in CA1 of the hippocampus that contains D-Cy5 (red in B, yellow where colocalized with green in C). This microglia extends processes that surround an unlabeled cell with a triangular, condensed nucleus (features typical of necrotic pyramidal cells in CA1) that also contains D-Cy5. (D–F) Microglia in the hippocampal dentate gyrus (DG) also contain D-Cy5 (arrowheads) and extend processes around adjacent unlabeled cells. (G–I) Astrocytes in CA1 were less often found to contain D-Cy5 (arrowheads). (Scale bar = 20 μm.) The Annals of Thoracic Surgery 2016 102, 743-750DOI: (10.1016/j.athoracsur.2016.02.077) Copyright © 2016 The Society of Thoracic Surgeons Terms and Conditions