murine sim a9 microglia  (ATCC)

Journal: Redox Biology

Article Title: Injectable ROS homeostasis protective hydrogel inhibiting microglial ferroptosis through the Nrf2/Slc7a11/Gpx4 to alleviate neuropathic pain and promote spinal cord injury repair

doi: 10.1016/j.redox.2025.103816

Figure Lengend Snippet: Preparation and characterization of MSQ and ROS scavenging of MSQ hydrogel (A)Nuclear magnetic resonance images of Galtin, Gel-SH, and Gel-MA. (B) FTIR spectra of quercetin, MS hydrogel, and MSQ hydrogel. (C)Swelling ratio of MS hydrogel and MSQ hydrogel. (n = 3) (D) SEM images of MS hydrogel and MSQ hydrogel, scale bar = 20μm/10 μm. (E, F) Rheological behaviors of MS hydrogel and MSQ hydrogel. (G) Photograph showing the injectability of the MSQ hydrogel. (H) Release curve of quercetin from MSQ hydrogel. (n = 3) (I) DPPH scavenging ratio of Gel-MA, MS hydrogel and MSQ hydrogel, compared with the control group, ∗P < 0.05, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. (n = 3) (J, K) DCFH-DA assay for ROS scavenging capacity of MS hydrogel and MSQ hydrogel in BV2 cells in vitro, scale bar = 25 μm, compared with the positive control group, ∗∗∗∗P < 0.0001. (n = 3).

Article Snippet: The immortalized BV2 murine microglia cells (CL-0493, Prooell Life Science & Technology Co., China) were cultured in complete culture medium that containing Dulbecco's modified Eagle's medium (DMEM; Invitrogen, Camarillo, CA), supplemented with 10 % (v/v) heat-inactivated fetal bovine serum (FBS, ExCell, FSP500), streptomycin (100 μg/mL), and penicillin (100 U/mL) at 37 °C under a humidified atmosphere with 5 % CO2.

Techniques: Nuclear Magnetic Resonance, Control, DCFH-DA Assay, In Vitro, Positive Control

Journal: Redox Biology

Article Title: Injectable ROS homeostasis protective hydrogel inhibiting microglial ferroptosis through the Nrf2/Slc7a11/Gpx4 to alleviate neuropathic pain and promote spinal cord injury repair

doi: 10.1016/j.redox.2025.103816

Figure Lengend Snippet: In vitro and in vivo biocompatibility of MSQ hydrogel (A) In vivo imaging of MSQ hydrogel implanted in mice, with images taken at 0-, 5-, 10-, and 14-days post-implantation. (B) The changes of the mean fluorescence intensity over time of MSQ hydrogel in vivo (n = 3). (C) In vitro degradation curves of MS and MSQ hydrogel. (D) Cell proliferation of BV2 cells treated with MS and MSQ hydrogel, assessed by CCK-8 assay (n = 3). (E) LDH release from BV2 cells treated with MS and MSQ hydrogel. (F, G) Live/dead staining of BV2 cells co-cultured with MS and MSQ hydrogel (scale bar = 50 μm) and corresponding statistical graphs. (n = 3) (H) Hemocompatibility of quercetin, MS and MSQ hydrogel. (n = 3).

Article Snippet: The immortalized BV2 murine microglia cells (CL-0493, Prooell Life Science & Technology Co., China) were cultured in complete culture medium that containing Dulbecco's modified Eagle's medium (DMEM; Invitrogen, Camarillo, CA), supplemented with 10 % (v/v) heat-inactivated fetal bovine serum (FBS, ExCell, FSP500), streptomycin (100 μg/mL), and penicillin (100 U/mL) at 37 °C under a humidified atmosphere with 5 % CO2.

Techniques: In Vitro, In Vivo, In Vivo Imaging, Fluorescence, CCK-8 Assay, Staining, Cell Culture

Journal: Redox Biology

Article Title: Injectable ROS homeostasis protective hydrogel inhibiting microglial ferroptosis through the Nrf2/Slc7a11/Gpx4 to alleviate neuropathic pain and promote spinal cord injury repair

doi: 10.1016/j.redox.2025.103816

Figure Lengend Snippet: Systems pharmacology analysis quercetin's microglial Nrf2 activation in post-SCI NP (A) Molecular docking results of Nrf2 with quercetin. (B) Root Mean Square Deviation (RMSD) results of Nrf2 with quercetin. (C) Radius of Gyration (Rg) results of Nrf2 with quercetin. (D–F) Immunofluorescence staining images and statistical graphs of Nrf2 in BV2 cells, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001, scale bar = 400 μm (n = 3) (E, G-I) Western blot analysis statistical graphs of nuclear Nrf2 and total Slc7a11/Gpx4 expressions in BV2 cells ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. (n = 3) (J–L) Immunofluorescence staining images and statistical graphs of JC-1 and FerroOrange in BV2 cells, ∗P < 0.05, ∗∗∗∗P < 0.0001, scale bar = 50 μm (n = 3) (M) 2D interaction diagram of quercetin-Nrf2. (N, P) Western blot analysis statistical graphs of nuclear Nrf2 expression in BV2 cells, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. (n = 4) (O, Q-R) Western blot analysis statistical graphs of total Slc7a11/Gpx4 expressions in BV2 cells, ∗P < 0.05, ∗∗P < 0.01. (n = 4).

Article Snippet: The immortalized BV2 murine microglia cells (CL-0493, Prooell Life Science & Technology Co., China) were cultured in complete culture medium that containing Dulbecco's modified Eagle's medium (DMEM; Invitrogen, Camarillo, CA), supplemented with 10 % (v/v) heat-inactivated fetal bovine serum (FBS, ExCell, FSP500), streptomycin (100 μg/mL), and penicillin (100 U/mL) at 37 °C under a humidified atmosphere with 5 % CO2.

Techniques: Activation Assay, Immunofluorescence, Staining, Western Blot, Expressing

Journal: Redox Biology

Article Title: Injectable ROS homeostasis protective hydrogel inhibiting microglial ferroptosis through the Nrf2/Slc7a11/Gpx4 to alleviate neuropathic pain and promote spinal cord injury repair

doi: 10.1016/j.redox.2025.103816

Figure Lengend Snippet: The impact of MSQ hydrogel on ferroptosis and inflammatory response in BV2 cells (A–B) Immunofluorescence staining images and statistical graphs of Nrf2 and Iba1 in BV2 cells, compared with Control group, ##P < 0.01, compared with LPS group, ∗P < 0.05, scale bar = 400 μm. (n = 3) (C-F) Western blot analysis and statistical graphs of nuclear Nrf2 and total Slc7a11 and Gpx4 expressions in BV2 cells, compared with Control group, #P < 0.05, compared with LPS group, ∗P < 0.05, ∗∗P < 0.01, compared with LPS + MSQ group, &P < 0.05. (n = 3) (G–J) Immunofluorescence staining images and statistical graphs of JC-1, FerroOrange and MitoSOX in BV2 cells, compared with Control group, ####P < 0.0001, compared with LPS group, ∗∗∗∗P < 0.0001, compared with LPS + MSQ group, &<0.05, &&P < 0.01, scale bar = 100 μm. (n = 3) (K–M) Determination of MDA, GSH, and Fe 2+ levels in microglial cells, compared with Control group, #P < 0.05, ###P < 0.001, ####P < 0.0001, compared with LPS group, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001, compared with LPS + MSQ group, &&P < 0.01, &&&&P < 0.0001. (n = 3) (N–Q) qPCR analysis of inflammatory factors expressions (TNFα and IL-1β) and M2 factors expressions (Arg1 and IL-4) in BV2 cells, compared with Control group, ##P < 0.01, ###P < 0.001, ####P < 0.0001, compared with LPS group, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001, compared with LPS + MSQ group, &P < 0.05, &&P < 0.01. (n = 3).

Article Snippet: The immortalized BV2 murine microglia cells (CL-0493, Prooell Life Science & Technology Co., China) were cultured in complete culture medium that containing Dulbecco's modified Eagle's medium (DMEM; Invitrogen, Camarillo, CA), supplemented with 10 % (v/v) heat-inactivated fetal bovine serum (FBS, ExCell, FSP500), streptomycin (100 μg/mL), and penicillin (100 U/mL) at 37 °C under a humidified atmosphere with 5 % CO2.

Techniques: Immunofluorescence, Staining, Control, Western Blot