Journal: Bioengineering

Article Title: Engineered Dual Antioxidant Enzyme Complexes Targeting ICAM-1 on Brain Endothelium Reduce Brain Injury-Associated Neuroinflammation

doi: 10.3390/bioengineering11030200

Figure Lengend Snippet: Representative images and quantitative evaluation of post-TBI neuropathological indices for neuronal cell death, astrocyte and microglial activation in animals treated with or without dual conjugates. Mice were placed in the following groups prior to brain harvesting and immunohistochemistry: naïve (craniotomy only but without impact), moderate CCI-TBI and moderate CC-TBI + 100 ng/kg of dual conjugate administered i.v. ( A ) NeuN staining identifies viable neurons within the cerebral cortex and lack of DAB+ staining for NeuN demonstrates the substantial neuronal loss at 48 h following CCI-TBI. The combination of mechanical strain, oxidative stress and inflammatory responses inevitably results in cell death. However, a rescue of viable neurons can be observed with the presence of dual conjugate administered soon after the TBI was induced. Astrocyte activation was determined by GFAP expression. CCI-TBI shows significant increase at 48 h following CCI-TBI, which is attenuated with the presence of the dual conjugate. Similarly, evaluation of activated microglial by phenotype and augmentation of expression of IBA-1 is clearly seen in the TBI animal. Dual conjugates also reduced the degree of microgliosis. Scale bar 50 microns ( B ) Bar graphs show the imaging quantification of the neuropathological indices from the groups outlined above. Data are presented as means ± SEM. For ( B – D ), one-way ANOVA with Dunnett’s test for multiple comparisons was used. ** p < 0.001, N.S = not significant.

Article Snippet: All sections were incubated in primary antibody prepared in Dako Antibody Diluent either for 1 h at RT (NeuN, Iba1) or overnight at 4 °C (GFAP) at the following dilutions: NeuN (1:500, Abnova Corp., Walnut, CA, USA), Iba-1 (1:400, (Fujifilm Wako Chemicals Inc., Richmond, VA, USA), GFAP (1:2000, Cell Signaling Technologies Inc., Danvers MA, USA).

Techniques: Activation Assay, Immunohistochemistry, Staining, Expressing, Imaging