Journal: Cell Death Discovery

Article Title: Preventing light-induced toxicity in a new mouse model of sector retinitis pigmentosa caused by Rhodopsin M39R variant

doi: 10.1038/s41420-025-02769-2

Figure Lengend Snippet: A Transcriptomic analysis by bulk RNA-seq of Rho M39R/M39R KI retinae compared to control mice. The volcano plot shows the top 35 differentially expressed genes by adjusted p value. S1P signalling pathway genes are underlined in red. fc_only = only fold change; not_sign = not significant; sig_only = only significant; sig+fc = significant plus fold change. B Rho M39R/+ KI mice were treated with FTY720 at 4 weeks of age. Schematic of the treatment in Rho M39R/+ KI mice. The animals were dark-adapted overnight and intraperitoneally injected with 10 mg/kg of FTY720 or vehicle (saline solution) 30 min before performing the light damage assay. The light damage consisted in performing an ERG every week for 4 times in total. Each time, the mice were preinjected with FTY720 or vehicle. C The ONL thickness was measured at day 21, after 4 rounds of ERG. FTY720-treated mice were compared to vehicle-treated and untreated mice. Mean ± SEM. Two-way ANOVA. Tukey’s multiple comparisons test between groups. (**** p < 0.0001, *** p < 0.001). Untreated N = 5, vehicle treated N = 7, FTY720 treated N = 6. D At the end of the light damage experiments, mice were culled, and the eyes were enucleated to perform further histological investigations. Rodents eyes were fixed in 4% PFA, incubated in 30% sucrose for 1–2 days, embedded in OCT (embedding matrix), cryosectioned and stained with DAPI. IHC of untreated, vehicle or FTY70 treated Rho M39R/+ superior retina after light damage. The cryosections were stained with rhodospin-4D2 (in magenta) and anti-GFAP (in yellow). Scale bar = 20 μm. E The % area occupied by GFAP-positive signal relative to the ONL, OPL, and INL was measured after thresholding GFAP staining in untreated and treated animals. Analysis performed in Fiji. Mean ± SEM. Two-way ANOVA. Tukey’s multiple comparisons test between groups. (** p < 0.001, *** p < 0.001). N = 3.

Article Snippet: Mice in dim red light were injected intraperitoneally with vehicle (saline solution) or FTY720 (Bio-techne, Tocris, 1 mg/ml in saline solution).

Techniques: RNA Sequencing, Control, Injection, Saline, Incubation, Staining

Journal: Cell Death Discovery

Article Title: Preventing light-induced toxicity in a new mouse model of sector retinitis pigmentosa caused by Rhodopsin M39R variant

doi: 10.1038/s41420-025-02769-2

Figure Lengend Snippet: A Rho M39R/M39R KI mice were treated with FTY720 at 4 weeks of age. Schematic of the FTY20 treatment in Rho M39R/M39R KI mice. Mice were dark-adapted and injected with FTY720 (10 mg/kg) 30 min prior to the light damage assay. The ERG was performed once to induce a faster degeneration. The OCT was performed at both day 0 and day 2, after 48 h from the single ERG. B The ONL thickness was measured at day 2, after a single ERG. FTY720-treated Rho M39R/M39R KI mice were compared to vehicle-treated and untreated mice. Mean ± SEM. Two-way ANOVA. Tukey’s multiple comparisons test between groups. (**** p < 0.0001, *** p < 0.001). Untreated N = 6, vehicle and FTY720 treated N = 5. C IHC of untreated, vehicle or FTY70 treated Rho M39R/M39R superior retina after light damage. The cryosections were stained with rhodospin-4D2 (in magenta). Scale bar = 20 μm. D The number of photoreceptors in the ONL was measured at 200–400 μm from the optic nerve in the inferior and superior retina. The analysis was performed on images of the central retina acquired with a microscope EVOS FL auto 2. The area of 10–20 nuclei per retina was measured and divided to total area of the ONL to calculate the total number of nuclei. The number of photoreceptors in 100 μm per treated/untreated animal was plotted. Mean ± SEM. Two-way ANOVA. Tukey’s multiple comparisons test between groups. (* p < 0.05, ** p < 0.01). N = 3.

Article Snippet: Mice in dim red light were injected intraperitoneally with vehicle (saline solution) or FTY720 (Bio-techne, Tocris, 1 mg/ml in saline solution).

Techniques: Injection, Staining, Microscopy

Journal: Bioactive Materials

Article Title: Modular assembled biomimetic nanobubbles for synergistic therapy of ischemic stroke via cascade modulation thrombo-inflammatory network

doi: 10.1016/j.bioactmat.2025.06.054

Figure Lengend Snippet: Schematic diagram of the modular assembly of PFNBs and their therapeutic mechanism in AIS. (A) The vicious cycle between BBB destruction and inflammatory response after AIS. (B) Selection of thrombo-inflammatory pathology adapted platelet membrane and FTY720 components, mechanism study of gas-liquid interface-mediated modular assembly and optimization. (C) The mechanism of PFNBs nano-bio interface-mediated AIS lesion targeting, BBB penetration enhancement, and the virtuous cycle establishment between anti-inflammatory regulation and BBB protection.

Article Snippet: Fingolimod (FTY720) hydrochloride was purchased from Selleck (Houston, TX, USA).

Techniques: Selection, Membrane

Journal: Bioactive Materials

Article Title: Modular assembled biomimetic nanobubbles for synergistic therapy of ischemic stroke via cascade modulation thrombo-inflammatory network

doi: 10.1016/j.bioactmat.2025.06.054

Figure Lengend Snippet: Fabrication, modular assembly mechanism and characterization of PFNBs. (A) Schematic diagram of the fabrication process of PFNBs. (B) Lipidomics characterization of the retention and content changes of phospholipid species in PLTs, PMVs and PNBs (n = 3). (C) Coarse-grained molecular dynamics simulation of the modular distribution of platelet membrane phospholipids during the reassembly process of the gas-liquid interface. (D) Coarse-grained molecular dynamics simulation of the modular distribution of platelet membrane phospholipids and gradient FTY720 contents during the reassembly process of the gas-liquid interface. The effect of gradient contents FTY720 embedding on phospholipid membrane fluidity (E) and intermolecular non-bonded interactions (F). (G) All-atom molecular dynamics simulation characterizes the assembly mode of FTY720 on the membrane shell of PFNBs. The image inserted in the red frame is an enlarged image of the FTY720 molecule. Particle size (H) and ζ potential characterization (I) of PFNBs loaded with gradient contents of FTY720 (n = 3). (J) TEM images of the PFNBs structure without negative staining (left) and with negative staining (right). Error bars: mean ± standard deviation.

Article Snippet: Fingolimod (FTY720) hydrochloride was purchased from Selleck (Houston, TX, USA).

Techniques: Membrane, Negative Staining, Standard Deviation

Journal: Bioactive Materials

Article Title: Modular assembled biomimetic nanobubbles for synergistic therapy of ischemic stroke via cascade modulation thrombo-inflammatory network

doi: 10.1016/j.bioactmat.2025.06.054

Figure Lengend Snippet: Composition and biofunction characterization of PFNBs. (A) The Venn diagram illustrates the same protein species overlap among PLTs, PMVs, and PFNBs. Volcano plot displaying differential proteins of PFNBs vs PMVs (B) and PFNBs vs PNBs (C). The red or blue plots represent significantly up-regulated or down-regulated proteins, respectively. (n = 3; fold change >2 and adj. P val < 0.05). Classification of PFNBs proteins by biological process (D) and molecular function (E). (F) Expression abundance of proteins associated with platelet-vascular injury targeting, intercellular adhesion, and immune escape properties in PLTs, PMVs, PNBs, and PFNBs (n = 3). (G) UV–Vis spectrophotometric detection of FTY720 encapsulation efficiency. (H) In vitro release profile of PFNBs (0.1 mg/mL, FTY720 solution concentrations) at different pH (n = 3). Error bars: mean ± standard deviation. ns indicates non-significant (p > 0.05). ∗p < 0.05 (two-tailed Student's t -test).

Article Snippet: Fingolimod (FTY720) hydrochloride was purchased from Selleck (Houston, TX, USA).

Techniques: Expressing, Encapsulation, In Vitro, Standard Deviation, Two Tailed Test

Journal: Bioactive Materials

Article Title: Modular assembled biomimetic nanobubbles for synergistic therapy of ischemic stroke via cascade modulation thrombo-inflammatory network

doi: 10.1016/j.bioactmat.2025.06.054

Figure Lengend Snippet: Evaluation of the immunomodulatory effects of PFNBs in vitro and in vivo . (A) Confocal fluorescence microscopy images of iNOS and CD206 immunofluorescence staining in inflammatory activated microglia after being incubated with Saline, free FTY720, PNBs, and PFNBs. Scale bar, 50 μm. (B) Quantitative analysis of iNOS (M1) and CD206 (M2) fluorescence intensity based on confocal images (n = 3). (C) WB bands of iNOS, CD206, p-STAT3 and t-STAT3 proteins. (D) Quantification of iNOS/GAPDH, CD206/GAPDH and p-STAT3/t-STAT3 levels,/Saline represents the change of experimental group relative to Saline group (n = 3). (E) Immunofluorescence staining and quantitative analysis of M1 (F) and M2 (G) phenotypes of microglia in the ischemic lesion (n = 5). Scale bar: 50 μm. (H) WB bands of iNOS, CD206, p-STAT3, and t-STAT3 proteins in lesion brain tissues of AIS mice. (I) Quantification of iNOS/GAPDH, CD206/GAPDH and p-STAT3/t-STAT3 levels in lesion brain tissues of AIS mice. Saline represents the change of experimental group relative to Saline group (n = 3). Error bars: mean ± standard deviation. ns indicates non-significant (p > 0.05). ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 (two-tailed Student's t -test).

Article Snippet: Fingolimod (FTY720) hydrochloride was purchased from Selleck (Houston, TX, USA).

Techniques: In Vitro, In Vivo, Fluorescence, Microscopy, Immunofluorescence, Staining, Incubation, Saline, Standard Deviation, Two Tailed Test

Journal: Bioactive Materials

Article Title: Modular assembled biomimetic nanobubbles for synergistic therapy of ischemic stroke via cascade modulation thrombo-inflammatory network

doi: 10.1016/j.bioactmat.2025.06.054

Figure Lengend Snippet: Evaluation of the protective effects on BBB permeability and vascular integrity of PFNBs in vitro and in vivo . (A) Schematic time line protocol of in vitro BBB model establishment and subsequent operations. (B) TEER value monitoring of endothelial cell monolayers after OGD/R treatment with addition of Saline, free FTY720, PNBs, and PFNBs (n = 3). (C) Proteomic analysis of the species and expression abundance of growth factors carried by platelet membranes in PLTs, PMVs, PNBs, and PFNBs (n = 3). (D) TEER value monitoring of endothelial cell monolayers seeded with inflammatory activated microglia in the lower chamber after OGD/R treatment with addition of Saline, free FTY720, PNBs, and PFNBs (n = 3). Confocal fluorescence microscopy images of CD34 and ZO-1 (E) or occluding (F) double-labeled immunofluorescence staining in the lesion area 24 h after AIS modeling and administration. Scale bar, 20 μm. Quantitative analysis of the ratio of ZO-1/CD34 positive area (G) or occludin/CD34 positive area (H) (n = 5). Photos of AIS mice brains in each group after Evans blue injection (I) and quantitative analysis of Evans blue leakage (J) (n = 3). (K) Quantitative analysis of brain edema in each group of AIS mice (n = 3). Error bars: mean ± standard deviation. ns indicates non-significant (p > 0.05). ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 (two-tailed Student's t -test).

Article Snippet: Fingolimod (FTY720) hydrochloride was purchased from Selleck (Houston, TX, USA).

Techniques: Permeability, In Vitro, In Vivo, Saline, Expressing, Fluorescence, Microscopy, Labeling, Immunofluorescence, Staining, Injection, Standard Deviation, Two Tailed Test

Journal: Bioactive Materials

Article Title: Modular assembled biomimetic nanobubbles for synergistic therapy of ischemic stroke via cascade modulation thrombo-inflammatory network

doi: 10.1016/j.bioactmat.2025.06.054

Figure Lengend Snippet: Neurobehavioral evaluation of mice in different groups (Sham, Saline, FTY720, PNBs, and PFNBs). (A) Schematic time line protocol of neurobehavioral evaluation. (B) The Modified Garcia Scores (including body proprioception, vibrissae touch, limb symmetry, lateral turning, forelimb walking and total neurological assessment score) in the different intervention groups at 24 h after stroke modeling (n = 6). Higher scores indicate better sensorimotor function performance. (C) Open field test and quantitative analysis average speed (D) and resting time (E). (F–I) Quantitative analysis of the rotarod test, wire hanging test, pole climbing test, and adhesion removal test of each group of mice during training, before modeling and 24 h after modeling and administration (n = 6). Error bars: mean ± standard deviation. ns indicates non-significant (p > 0.05). ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 (two-tailed Student's t -test).

Article Snippet: Fingolimod (FTY720) hydrochloride was purchased from Selleck (Houston, TX, USA).

Techniques: Saline, Modification, Standard Deviation, Two Tailed Test