Additional file 1 of An injectable refrigerated hydrogel for inducing local hypothermia and neuroprotection against traumatic brain injury in mice
Creators: name: Han, Yuhan
name: Han, Zhengzhong
name: Huang, Xuyang
name: Li, Shanshan
name: Jin, Guoliang
name: Feng, Junfeng
name: Wu, Decheng
name: Liu, Hongmei
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: Neuroscience
subject: Pharmacology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Science Policy

description: Additional file 1: Figure S1. The heating coil installation and temperature measurement. Figure S2. Average levels of T1AM in cerebral hemisphere measured by HPLC‐MS/MS. Figure S3. The security of the Pol hydrogel by measuring cell viability. Figure S4. Average levels of T1AM in hypothalamus measured by HPLC‐MS/MS. Figure S5. Survival curve for the mice treated with different dose of T1AM, intraperitoneal injection. (n = 5 mice). Figure S6. Body and brain temperature of the male and female mice treated with the Pol/T hydrogel in WDI models. Figure S7. The temperature variation in the brain of mice in sham model received the Pol/T and Pol treatments along with the time. Figure S8. The hemoglobin content of brain after 12 h post-TBI in different groups. Figure S9. (A) Expression of Tunel, Bcl-2, Bax, MMP9 in brain tissue at 21 d. Scale bar = 100 μm. (B) Number of Tunel + cell death/field (a), Bcl-2 positive cells/field (b), Bax positive cells/field (c) and MMP9 positive cells/field (d). (B) Data were presented as means ± SDs (5 technical replicates averaged for each 3 mice/group). Normality and homogeneity of variance was checked using Shapiro–Wilk test. Statistical significance of the results was determined using one-way ANOVA with Tukey’s post hoc test. Figure S10. Brain section from TBI mice, taken at day 21 post-injury, with indicated treatments administered at 21 post-injury time point. Figure S11. (A) Representative fields of cells positive for TdT-mediated dUTP nick-end labeling (Tunel) stain in the different groups. Scale bar = 50 μm. (B) Quantified bar graph of Tunel + cell death. ****P &lt; 0.0001 were performed using one-way ANOVA with Tukey’s post hoc test. Figure S12. (A) The ratio of Bcl-2/Bax-positive cells/field in WDI models. (B) The ratio of Bcl-2/Bax-positive cells/field in PBI models. Data were represented as means ± SDs (n = 15 fields of 3 mice). *P &lt; 0.05, **P &lt; 0.01 and ****P &lt; 0.0001 were performed using Kruskal–Wallis test. Figure S13. (A) Representative fields of cells positive for Tunel stain in the different groups. Scale bar = 50 μm. (B) Quantified bar graph of Tunel + cell death. (C) Representative fields of Bcl-2 staining around the injury site in different groups (a) Representative fields of Bax staining around the injury site in different groups (b). Scale bar = 100 μm. (D) The ratio of Bcl-2/Bax-positive cells/field. (B) Data are represented as means ± SDs (5 technical replicates averaged for each 3 mice/group). **P &lt; 0.01 and ****P &lt; 0.0001 were performed using one-way ANOVA with Tukey’s post hoc test. (D) Data are represented as means ± SDs (5 technical replicates averaged for each 3 mice/group). **P &lt; 0.01, ***P &lt; 0.001 and ****P &lt; 0.0001 were performed using Kruskal–Wallis test. Figure S14. Local hypothermia induced by the Pol/T hydrogel protected the integrity of the BBB and educed brain edema in the TBI mouse model. Figure S15. Brain water content. Data were presented as means ± SDs (n = 5 mice). Normality was checked using Shapiro–Wilk test. *P &lt; 0.05, **P &lt; 0.01 and ****P &lt; 0.0001 were performed using one-way ANOVA with Tukey’s post hoc test. Figure S16. Levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6) and chemokine1 (CXCL1) in injured tissue by enzyme linked immunosorbent assay (ELISA) at 12 h after TBI. Data were presented as means ± SD (n = 3 mice). Figure S17. Analysis of neuroinflammation in response to different treatments. Figure S18. Functional recovery assessments by Morris water maze test, mNSS score and wire hanging test on 21 d after TBI. Figure S19. Brain temperature variation of Sham group, T1AM group and T1AM + EPPTB group. Data were presented as means ± SDs (n = 4 mice).