Journal: Bioengineering & Translational Medicine

Article Title: Neuron‐targeted 2‐deoxyglucose‐dendrimer‐rosiglitazone nanotherapy mitigates neuroinflammation and cognitive deficits in pediatric traumatic brain injury

doi: 10.1002/btm2.70053

Figure Lengend Snippet: FIGURE 8 mRNA expressions of pro- and anti-inflammatory markers and cell death markers from sham (n = 12, 6 M/6 F), traumatic brain injury (TBI) + vehicle (n = 12, 6 M/6 F), TBI + rosiglitazone (Rosi) (n = 12, 6 M/6 F), and TBI + 2-deoxyglucose (2DG-D)-Rosi (n = 12, 6 M/6 F) groups. Neurons were isolated from the injured brain regions (or the matching area from the sham mice) at 24-h post-treatment for gene expression evaluations. Data were presented as mean ± SEM. Treatment groups were presented as sham (gray circles), TBI + vehicle (blue circles), TBI + Rosi (magenta circles), and TBI + 2DG-D-Rosi (green circles). (a–d) The expression of pro-inflammatory makers, tumor necrosis factor-alpha (TNF-α) (a), interleukin-6 beta (IL-1β) (b), Toll-like receptor 4 (TLR4) (c), and NLR Family Pyrin Domain Containing 3 (NLRP3) (d). (e) The expression of anti-inflammatory maker IL-13. (f) The expression of cell death marker Fas. *p < 0.05; **p < 0.01; ***p < 0.001. (g) The co- localization of neurodegenerative markers marker Fluoro-Jade C (FJC) (green) and neuronal marker (NeuN, red) was evaluated in sham (n = 8, 4 M/4 F), TBI + vehicle (n = 8, 4 M/4 F), TBI + Rosi (n = 8, 4 M/4 F), and TBI + 2DG-D-Rosi (n = 8, 4 M/4 F) groups at 24-h post-injury. Images (40, 5 images/animal) were randomly acquired from the cortex area (mainly primary motor cortex and primary somatosensory cortex) in the injured brain regions (approximately between bregma +1 mm and bregma 0.5 mm) using the Nikon Eclipse TS2R fluorescent microscope (Nikon, NY, USA). The representative images from male treatment groups (upper panels) and female treatment groups (lower panels). 40,6-Diamidino-2-phenylindole (DAPI) (blue) was used for nucleus stains. Scale bars: 100 μm. (h) The quantification of FJC+NeuN+ cells among treatment groups. Data were presented as mean ± SEM. Treatment groups were presented as sham (gray circles), TBI + vehicle (blue circles), TBI + Rosi (magenta circles), and TBI + 2DG-D-Rosi (green circles). *p < 0.05; **p < 0.01; ****p < 0.0001.

Article Snippet: Brain sections were washed 3 times in PBS for 5 min each, incubated in 0.001% Fluoro-Jade C (FJC) (Cat# TR-160-FJC, Biosensis, CA, USA) for 10 min, washed six times in PBS for 15 min each, and incubated with rabbit anti-NeuN (1:250; Cat# ab177487; Abcam, MA, USA).

Techniques: Isolation, Gene Expression, Expressing, Marker, Microscopy