Journal: iScience

Article Title: Fabrication of RADA32/Ngf_EE/MSCs composite hydrogel and its protective mechanism against radiation-induced ototoxicity

doi: 10.1016/j.isci.2026.115723

Figure Lengend Snippet: RNM composite gel mitigates radiation-induced damage in vitro (A) Viability of HEI-OC1 cells after different doses of radiation treatment, detected by CCK-8 assay. (B) Apoptosis of HEI-OC1 cells analyzed by flow cytometry after radiation exposure and subsequent transwell co-culture with RN gel, MSCs, or the RNM gel system. Data are represented as the mean ± SEM. ( N = 3, t test). (C) Distribution of γ-H2AX (red) in HEI-OC1 cells observed under a confocal microscope after radiation exposure and intervention with RN, MSCs, or RNM. Scale bars, 10 μm. (D, E, G, and H) mRNA expression levels of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) in mouse cochlear tissues evaluated by real-time qPCR after intratympanic injection of RN, MSCs, or RNM hydrogel following radiation exposure. Data are represented as the mean ± SEM ( N = 3, t test). (F) Intracellular ROS levels in HEI-OC1 cells monitored using the DCFH-DA fluorescent probe and flow cytometry after radiation exposure and intervention with RN, MSCs, or RNM. Data are represented as the mean ± SEM ( N = 3, t test). (I) mRNA expression levels of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) in HEI-OC1 cells detected by real-time qPCR after radiation exposure and intervention with RN, MSCs, or RNM. Data are represented as the mean ± SEM ( N = 3, t test). Significant differences between the groups are indicated by ∗ for p < 0.05, ∗∗ for p < 0.01, ∗∗∗ for p < 0.001, and ∗∗∗∗ for p < 0.0001.

Article Snippet: Mouse bone marrow-derived mesenchymal stem cells (MSCs) and the mouse monocyte/macrophage cell line RAW264.7 were purchased from Procell Life Science & Technology Co., Ltd.

Techniques: In Vitro, CCK-8 Assay, Flow Cytometry, Co-Culture Assay, Microscopy, Expressing, Injection

Journal: iScience

Article Title: Fabrication of RADA32/Ngf_EE/MSCs composite hydrogel and its protective mechanism against radiation-induced ototoxicity

doi: 10.1016/j.isci.2026.115723

Figure Lengend Snippet: In vivo protective effects of RNM composite gel (A) Schematic of the in vivo experimental timeline and anatomical images of the mouse cochlea under a dissection microscope. (a, oval window; b, cochlear labyrinth.). (B–G) ABR tests performed 2 days before radiation exposure and 3, 7, and 14 days after drug administration to assess hearing function in mice. (H) Left: Confocal microscopy images of the cochlear basilar membrane 14 days after radiation exposure and intratympanic injection of RN, MSCs, or RNM hydrogel, stained with FITC-phalloidin to label hair cells. Scale bars, 20 μm. Right: Quantitative comparison of outer hair cell (OHC) and inner hair cell (IHC) survival rates post-intervention. Data are represented as the mean ± SEM ( N = 3, t test). (I) H&E staining images of mouse cochlear tissues extracted 14 days after radiation exposure and intratympanic injection of RN, MSCs, or RNM hydrogel. Data are represented as the mean ± SEM ( N = 3, t test). Scale bars, 20 μm. Significant differences between the groups are indicated by ∗ for p < 0.05, ∗∗ for p < 0.01, ∗∗∗ for p < 0.001, and ∗∗∗∗ for p < 0.0001.

Article Snippet: Mouse bone marrow-derived mesenchymal stem cells (MSCs) and the mouse monocyte/macrophage cell line RAW264.7 were purchased from Procell Life Science & Technology Co., Ltd.

Techniques: In Vivo, Dissection, Microscopy, Confocal Microscopy, Membrane, Injection, Staining, Comparison

Journal: iScience

Article Title: Fabrication of RADA32/Ngf_EE/MSCs composite hydrogel and its protective mechanism against radiation-induced ototoxicity

doi: 10.1016/j.isci.2026.115723

Figure Lengend Snippet: Immunomodulatory mechanism of RNM composite gel (A and B) Flow cytometric analysis of CD86 and CD206 expression in RAW264.7 macrophages after irradiation and co-culture with RN, MSCs, or RNM composite gel in a transwell system (macrophages in lower chamber). Data are represented as the mean ± SEM ( N = 3, t test). (C) Immunofluorescence staining of F4/80 (red) on cochlear sections. Scale bars, 50 μm (a, spiral ganglion; b, basilar membrane; c, stria vascularis; d, spiral ligament). (D) Apoptosis of HEI-OC1 cells analyzed by flow cytometry after radiation exposure and intervention. Data are represented as the mean ± SEM ( N = 3, t test). (E) Expression level of p-p65, a key marker of NF-κB pathway activation, in macrophages after radiation exposure and drug intervention. Data are represented as the mean ± SEM ( N = 3, t test). Significant differences between the groups are indicated by ∗ for p < 0.05, ∗∗ for p < 0.01, ∗∗∗ for p < 0.001, and ∗∗∗∗ for p < 0.0001.

Article Snippet: Mouse bone marrow-derived mesenchymal stem cells (MSCs) and the mouse monocyte/macrophage cell line RAW264.7 were purchased from Procell Life Science & Technology Co., Ltd.

Techniques: Expressing, Irradiation, Co-Culture Assay, Immunofluorescence, Staining, Membrane, Flow Cytometry, Marker, Activation Assay