Journal: Advanced Science

Article Title: Remote Activation of Spinal TRPV1 by Magnetic Nanocubes Confers Cardioprotection Against Myocardial Ischemia‐Reperfusion Injury

doi: 10.1002/advs.202520852

Figure Lengend Snippet: Preparation and characterization of FeNCs‐TRPV1. A) Schematic diagram of the synthesis of FeNCs‐TRPV1. B,C) Transmission electron microscopy images of FeNCs and FeNCs‐TRPV1. Scale bar = 50 nm. D) Zeta potential results of FeNCs and FeNCs‐TRPV1. Data represent the mean ± standard deviation (SD) from three independent experiments. Statistical analysis was performed using an unpaired t ‐test. E) Infrared thermal images of FeNCs‐TRPV1 (0.1 mg mL −1 ) upon exposure to ACMF at different magnetic densities. F) Infrared thermal images of FeNCs‐TRPV1 at different concentrations upon exposure to ACMF at 15 A. G) Optical fiber thermometer recording of FeNCs‐TRPV1 (0.1 mg mL −1 ) upon exposure to ACMF at different magnetic densities. H) Optical fiber thermometer recording of FeNCs‐TRPV1 at different concentrations upon exposure to ACMF at 15 A. Δ T : Temperature rise. Data represent the mean ± SD from four independent experiments in G and H, and statistical analysis was performed using repeat measurement two‐way analysis of variance (ANOVA) followed by Tukey's test. ** p <0.01.

Article Snippet: F11 cells successfully transfected with the rTRPV1 plasmid were cultured for 48 h, then divided into 3 groups and incubated with extracellular TRPV1 antibody (TRPV1 Ab, ACC‐029, Alomone), FeNCs‐TRPV1, or PBS for 2 h. Meanwhile, cells transfected with the empty vector were set as the control group.

Techniques: Transmission Assay, Electron Microscopy, Zeta Potential Analyzer, Standard Deviation

Journal: Advanced Science

Article Title: Remote Activation of Spinal TRPV1 by Magnetic Nanocubes Confers Cardioprotection Against Myocardial Ischemia‐Reperfusion Injury

doi: 10.1002/advs.202520852

Figure Lengend Snippet: Specific binding of FeNCs‐TRPV1 to TRPV1‐expressed F11 cells. A) Cell viability of F11 cells incubated with FeNCs or FeNCs‐TRPV1 at different iron concentrations for 24 h. B) Cell viability of F11 cells incubated with FeNCs or FeNCs‐TRPV1 at the iron concentration of 0.1 mg mL −1 for 24, 48, and 72 h. PBS served as a control. Data represent the mean ± SD of four independent experiments and were analyzed with the two‐way ANOVA followed by Tukey's test. C) Prussian blue staining images of the transfected F11 cells incubated with FeNCs or FeNCs‐TRPV1 for 2 h. Red arrows point to blue‐stained iron nanocubes. Scale bar = 20 µm. D) Representative transmission electron microscopy images of F11 cells incubated with 0.1 mg mL −1 FeNCs‐TRPV1 or FeNCs for 2 h. The white dotted outline represents the cell membrane. Right panels are enlarged images showing cell surface area (I) and intracellular area (II). Red arrows indicate the localization of nanocubes. Scale bar = 2 µm.

Article Snippet: F11 cells successfully transfected with the rTRPV1 plasmid were cultured for 48 h, then divided into 3 groups and incubated with extracellular TRPV1 antibody (TRPV1 Ab, ACC‐029, Alomone), FeNCs‐TRPV1, or PBS for 2 h. Meanwhile, cells transfected with the empty vector were set as the control group.

Techniques: Binding Assay, Incubation, Concentration Assay, Control, Staining, Transfection, Transmission Assay, Electron Microscopy, Membrane

Journal: Advanced Science

Article Title: Remote Activation of Spinal TRPV1 by Magnetic Nanocubes Confers Cardioprotection Against Myocardial Ischemia‐Reperfusion Injury

doi: 10.1002/advs.202520852

Figure Lengend Snippet: FeNCs‐TRPV1 promotes TRPV1‐mediated Ca 2+ influx through magnetothermal effect. A) Representative image of F11 cells under patch clamp recording. Scale bar = 25 µm. B) Representative traces of capsaicin‐induced inward currents in TRPV1‐transfected F11 cells incubation with PBS (gray), TRPV1 antibody (TRPV1 Ab, blue), or FeNCs‐TRPV1 (red). The vector‐transfected F11 cells served as the blank control (black). C) Quantitative analysis of the amplitude of capsaicin‐induced inward currents. Data represent the mean ± SD. Statistical analysis was performed using one‐way ANOVA followed by Tukey's test ( n = 9 cells per group from three independent experiments). D,E) Representative images for intracellular Ca 2+ signal detected by Fura‐4AM fluorescent probe in F11 cells (D) or primary DRG neurons (E). Scale bar = 50 µm. F) Fluorescent intensity (arbitrary unit, A.U.) was quantified based on four independent experiments. G) Representative immunoreactive bands of phosphorylated Camkk2 (p‐Camkk2), Camkk2, phosphorylated AMPK (p‐AMPK), AMPK, and the reference gene GAPDH. H) The relative levels of p‐Camkk2 and p‐AMPK were expressed as ratios of their phosphorylated forms to total protein, following normalization to GAPDH, and the value in the FeNCs‐treated vector group was assigned as 1. Data represent the mean ± SD from four independent experiments. The analysis was performed using the two‐way analysis of ANOVA followed by Tukey's test.

Article Snippet: F11 cells successfully transfected with the rTRPV1 plasmid were cultured for 48 h, then divided into 3 groups and incubated with extracellular TRPV1 antibody (TRPV1 Ab, ACC‐029, Alomone), FeNCs‐TRPV1, or PBS for 2 h. Meanwhile, cells transfected with the empty vector were set as the control group.

Techniques: Patch Clamp, Transfection, Incubation, Plasmid Preparation, Control

Journal: Advanced Science

Article Title: Remote Activation of Spinal TRPV1 by Magnetic Nanocubes Confers Cardioprotection Against Myocardial Ischemia‐Reperfusion Injury

doi: 10.1002/advs.202520852

Figure Lengend Snippet: FeNCs‐TRPV1 mitigates myocardial injury in rats subjected to repeated ACMF exposure. A) Schematic diagram for intra‐spinal injection of nanocubes. B) Prussian blue staining of the longitudinally transected spinal cord. The red arrows point to the blue‐stained nanocubes. Scale bar = 500 µm. C) MRI images of the spinal cord. The white dotted outline represents the area of the spinal cord. The red arrow points to the localization of the injected nanocubes. D) Schematic diagram of the experimental procedure. E) Representative images of heart sections stained by TTC (2,3,5‐triethylenetetrazolium chloride) and Evans blue. F) Infarct size (IS) is expressed as the ratio to the area at risk (AAR). G) The ratio of AAR to left ventricular (LV) and right ventricular (RV). H) Serum cardiac troponin I (cTnI) levels measured by ELISA. Data were expressed as the mean ± SD and were analyzed using two‐way ANOVA followed by Tukey's test ( n = 6 per group).

Article Snippet: F11 cells successfully transfected with the rTRPV1 plasmid were cultured for 48 h, then divided into 3 groups and incubated with extracellular TRPV1 antibody (TRPV1 Ab, ACC‐029, Alomone), FeNCs‐TRPV1, or PBS for 2 h. Meanwhile, cells transfected with the empty vector were set as the control group.

Techniques: Injection, Staining, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Remote Activation of Spinal TRPV1 by Magnetic Nanocubes Confers Cardioprotection Against Myocardial Ischemia‐Reperfusion Injury

doi: 10.1002/advs.202520852

Figure Lengend Snippet: FeNCs‐TRPV1 alleviates ventricular arrhythmia and promotes cardiac cells survival. A) Arrhythmia score for each group. B) Serum Norepinephrine levels measured by ELISA for each group. C) Representative immunoreactive bands of p‐Akt/Akt, p‐ERK1/2/ERK1/2, p‐GSK‐3β/GSK‐3β, and the reference GAPDH in heart tissue samples. D–F) The relative levels of p‐Akt, p‐ERK1/2, and p‐GSK‐3β were expressed as ratios of their phosphorylated forms to total protein, following normalization to GAPDH, and the value in the FeNCs‐injected Sham group was assigned as 1. G) Representative immunoreactive bands of Bax/Bcl‐2, cleaved caspase‐3/caspase‐3, and the reference GAPDH in heart tissue samples. H,I) The relative ratios of Bax/Bcl‐2 and cleaved caspase‐3/caspase‐3 were calculated following normalization to GAPDH, the value in the FeNCs‐injected Sham group was assigned as 1. J) Representative images for TUNEL staining in the left ventricular tissues. Scale bar = 20 µm. K) The number of TUNEL‐positive apoptotic cells was quantified and expressed as a percentage of the total number of cells. Data represent the mean ± SD. Statistical analysis was performed using two‐way ANOVA followed by Tukey's test ( n = 6 per group in A, B, n = 5 per group in C–I, n = 6 per group in J,K).

Article Snippet: F11 cells successfully transfected with the rTRPV1 plasmid were cultured for 48 h, then divided into 3 groups and incubated with extracellular TRPV1 antibody (TRPV1 Ab, ACC‐029, Alomone), FeNCs‐TRPV1, or PBS for 2 h. Meanwhile, cells transfected with the empty vector were set as the control group.

Techniques: Enzyme-linked Immunosorbent Assay, Injection, TUNEL Assay, Staining

Journal: Advanced Science

Article Title: Remote Activation of Spinal TRPV1 by Magnetic Nanocubes Confers Cardioprotection Against Myocardial Ischemia‐Reperfusion Injury

doi: 10.1002/advs.202520852

Figure Lengend Snippet: FeNCs‐TRPV1 inhibits calcium‐dependent signaling and neuropeptides release in the spinal cord. A) Representative immunoreactive bands of p‐Camkk2/Camkk2, p‐AMPK/AMPK, and the reference GAPDH in spinal cord tissue samples. B,C) The relative levels of p‐Camkk2 and p‐AMPK were expressed as ratios of their phosphorylated forms to total protein, following normalization to GAPDH, and the value in the FeNCs‐injected Sham group was assigned as 1. D) Representative images showing CGRP and SP immunostaining in the spinal cord issues from rats. Scale bar = 50 µm. E,F) Quantification of CGRP (E) and SP (F) fluorescence intensity (A.U.). Data represent the mean ± SD. Statistical analysis was performed using two‐way ANOVA followed by Tukey's test ( n = 4 per group).

Article Snippet: F11 cells successfully transfected with the rTRPV1 plasmid were cultured for 48 h, then divided into 3 groups and incubated with extracellular TRPV1 antibody (TRPV1 Ab, ACC‐029, Alomone), FeNCs‐TRPV1, or PBS for 2 h. Meanwhile, cells transfected with the empty vector were set as the control group.

Techniques: Injection, Immunostaining, Fluorescence

Journal: Advanced Science

Article Title: Remote Activation of Spinal TRPV1 by Magnetic Nanocubes Confers Cardioprotection Against Myocardial Ischemia‐Reperfusion Injury

doi: 10.1002/advs.202520852

Figure Lengend Snippet: FeNCs‐TRPV1 mediates spinal TRPV1 channel desensitization upon repeated ACMF exposure. A) Schematic diagram of the experimental procedure for 1, 2, or 3 cycles of ACMF exposure without subsequent myocardial IR. B) Representative immunoreactive bands of p‐AMPK/AMPK, p‐Camkk2/Camkk2, and the reference GAPDH in spinal cord tissue samples. C,D) The relative levels of p‐Camkk2 and p‐AMPK were expressed as ratios of their phosphorylated forms to total protein, following normalization to GAPDH, and the value in the FeNCs‐injected Control group was assigned as 1 ( n = 5 per group). E) Schematic diagram of the experimental procedure for 1, 2, or 3 cycles of ACMF exposure with subsequent myocardial IR. F) Representative immunoreactive bands of p‐Camkk2/Camkk2, p‐AMPK/AMPK, and the reference GAPDH in spinal cord tissue samples. G,H) The relative levels of p‐Camkk2 and p‐AMPK were expressed as ratios of their phosphorylated forms to total protein, following normalization to GAPDH, and the value in FeNCs‐injected rats subjected to 1 cycle of ACMF was assigned as 1 ( n = 5 per group). I) Representative images showing CGRP and SP immunostaining in the spinal cord issues from rats. Scale bar = 50 µm. J,K) Quantification of CGRP (J) and SP (K) fluorescence intensity (A.U.). Data represent the mean ± SD. Statistical analysis was performed using two‐way ANOVA followed by Tukey's test ( n = 5 per group in B–D and F–H, n = 4 per group in I–K).

Article Snippet: F11 cells successfully transfected with the rTRPV1 plasmid were cultured for 48 h, then divided into 3 groups and incubated with extracellular TRPV1 antibody (TRPV1 Ab, ACC‐029, Alomone), FeNCs‐TRPV1, or PBS for 2 h. Meanwhile, cells transfected with the empty vector were set as the control group.

Techniques: Injection, Control, Immunostaining, Fluorescence

Journal: Advanced Science

Article Title: Remote Activation of Spinal TRPV1 by Magnetic Nanocubes Confers Cardioprotection Against Myocardial Ischemia‐Reperfusion Injury

doi: 10.1002/advs.202520852

Figure Lengend Snippet: A summary figure illustrating that FeNCs‐TRPV1 exerts cardioprotective effects upon exposure to ACMF through magnetothermal effects. When the FeNCs‐TRPV1 nanocubes were injected into the rat spinal cord, repetitive and transient ACMF exposure induced the intermittent activation of the TRPV1 channel, which induced TRPV1 desensitization and ultimately prevented the hyperactivation of TRPV1 during the subsequent ischemia and reperfusion phases. This resulted in a significant reduction of myocardial ischemic injury in rats, and the cardioprotection is potentially associated with the reduced release of neuropeptides, decreased cardiac sympathetic efferent activity, enhanced intramyocardial prosurvival signaling, and the inhibition of apoptotic signaling. The summary figure was originally created by the authors and professionally refined by a scientific illustration service. The copyright is retained by the authors.

Article Snippet: F11 cells successfully transfected with the rTRPV1 plasmid were cultured for 48 h, then divided into 3 groups and incubated with extracellular TRPV1 antibody (TRPV1 Ab, ACC‐029, Alomone), FeNCs‐TRPV1, or PBS for 2 h. Meanwhile, cells transfected with the empty vector were set as the control group.

Techniques: Injection, Activation Assay, Activity Assay, Inhibition