A therapeutic relay strategy enabled by spatiotemporally programmable nanoplatforms for multilayered cardioprotection against MI/RI

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  Scheme 1. Schematic illustration of a spatiotemporally programmable nanoplatform integrating for multilayered cardioprotection against MI/RI. MP@T NPs was synthesized using thin film hydration method and phacoemulsification method. The TA-Ce network exhibits dual anti-inflammatory and endogenous ROS-scavenging capacities. US stimulation exerts a specific regulatory effect at twenty-four hours following ischemia-reperfusion injury, and ferroptosis initiates its onset phase

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  Fig. 1. Bioinformatics analysis, Synthesis, Characterization and Enzymatic activity of MP@T NPs. (A) Different expression of GPX4 between control and disease stissues in the GSE83472 database. (B) GSEA analysis. (C) Circos plot of the top 10 genes positively/negatively correlated with GPX4 expression. (D) Correlation analysis between GPX4 and DYRK1B. (E) TEM images of MP NPs, scale bar is 100 nm. (F) TEM images of MP@T NPs, scale bar is 200 nm (inset: scale bar is 100 nm). (G) The hydrodynamic sizes of MP NPs and MP@T NPs. (H) Zeta potentials of MP NPs and MP@T NPs. (I) UV-vis spectra of PFP@lip NPs, TA-Ce, MP NPs and MP@T NPs. XPS patterns of MP NPs and MP@T NPs (J) and high-resolution XPS patterns of Ce 3d (K). (L) The FTIR spectra of MP NPs, TA-Ce and MP@T NPs. (M) Schematic illustration shows enzymatic activity of MP@T NPs. (N) O2 generation after different treatments as measured by a dissolved oxygen meter. (O) H2O2 scavenging ability with different concentrations. (P) O2- inhibition rate with different concentrations. (Q) •OH scavenging ability with different concentrations. (R-S) EPR spectra analysis of O2- and •OH scavenging with MP@T NPs. (T) Total antioxidant capacity of MP@T…

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  Fig. 2. Efficient cellular targeting and ROS scavenging in vitro. Relative cell viabilities of H9c2 cells (A) and HUVEC (B) incubated with MP@T NPs at varied concentrations for 24 h. (C) ICP results of H9c2 cells treated with MP@T NPs for different time. Fluorescence image (D) and (E) flow cytometry results of H9c2 cells treated with Dil-labeled MP NPs and Dil-labeled MP@T NPs for different time, scale bar is 100 μm. Flow cytometry results of DCFH-DA (F), RDPP (G) and MitoSox (H) in H9c2 cells after different treatments. (I) Fluorescence image of DCFH-DA, RDPP and MitoSox in H9c2 cells after different treatments, scale bar is 100 μm. Flow cytometry results of fluorescence (FL) intensity (J) and MFI quantitative analysis (K) of DCFH-DA in H9c2 cells after different treatments. Flow cytometry results of fluorescence (FL) intensity (L) and MFI quantitative analysis (M) of RDPP in H9c2 cells after different treatments. Flow cytometry results of fluorescence (FL) intensity (N) and MFI quantitative analysis (O) of MitoSox in H9c2 cells after different treatments. Data were presented as mean ± SD, n = 3. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, no significance

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  Fig. 3. Protective effect of MP@T NPs on OGD/R injured H9c2 cells. (A) Schematic diagram of protective effect of MP@T NPs. (B) Color division diagram of different groups. Relative cell viabilities of H9c2 cells treated with US irradiation before (C) and after OGD/R (D). Quantitative analysis (E) and fluorescence image (F) of BODIPY 581/591 C11 in H9c2 cells after different treatments, scale bar is 100 μm. Lipid peroxidation levels (G), and GPx activity (H) of H9c2 cells subjected to different treatments. Western blot bands (I) and quantitative analysis of GPX4 (J) and ACSL4 (K) expression. (L) TEM images of H9c2 cells subjected to different treatments (scale bar: 500 nm). (M) Analysis of mitochondrial membrane potential of H9c2 cells after different treatments using JC-1 staining. (N) Quantitative ratio analysis of aggregated and monomeric JC-1. (O) ATP productions of H9c2 cells subjected to different treatments. (P) PCA analysis of mRNA of H9c2 cells after treated with OGD/R or MP@T + US. (Q) Volcano plot of differently expressed genes of H9c2 cells after treated with OGD/R or MP@T + US (n = 5). (R) Analysis of KEGG enrichment pathways for OGD/R group and MP@T NPs + US group. Data were…

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  Fig. 4. Targeting ability of MP@T NPs and the ultrasonic profile in MI/RI rats. Accumulations of DiR-labeled MP NPs and DiR-labeled MP@T NPs in heart (A) and main organs (B) in different time points of administration. (C) Fluorescence image Dil-labeled MP NPs and Dil-labeled MP@T NPs in heart in 3 h of administration, scale bar is 5 mm. Calculated results of heart (D), liver (E), spleen (F), lung (G), and kidney (H). (I) The US imaging (left: B-Mode, right: CEUS) of MP@T NPs with time-and intensity-dependent ADV. Quantitative analysis of the echo intensity of MP@T NPs after US irradiation at different intensities and time in B-mode (J) and CEUS-mode (K).In vivo US imaging (up: B-Mode, down: CEUS) of hearts in model rat pre-operation, baseline, and after US irradiation. Data were presented as mean ± SD, n = 3. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, no significance

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  Fig. 5. Efficacy evaluations of acute MI/RI injury alleviation by MP@T NPs in vivo. (A) Schematic illustration of acute MI/RI model establishment and experiment schedule. (B) Image of Perls (scale bar: 200 μm), fluorescence image of DHE (scale bar: 50 μm), TNF-α (scale bar: 50 μm), IL-10 (scale bar: 50 μm), and TEM images (scale bar: 2 μm and 500 nm) after different treatments. Quantitative analysis of Perls (C), DHE (D), TNF-α (E) and IL-10 (F). (G) Heat map of inflammatory genes in heart tissue after treatment of MI/RI or MP@T NPs (n = 3). (H) Analysis of GO enrichment pathways for MI/RI or MP@T NPs group. (I-K) Level of inflammatory cytokines (IL-10, IL-1β and IL-6) in plasma of MI/RI rats after different treatments. (L-M) Level of CK and LDH in plasma of MI/RI rats after different treatments. Data were presented as mean ± SD, n = 5. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, no significance

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  Fig. 6. Efficacy evaluations of chronic MI/RI injury alleviation by MP@T NPs in vivo. (A) Schematic illustration of chronic MI/RI model establishment and experiment schedule. (B) Representative HE (scale bar: 2 mm and 200 μm), masson (scale bar: 200 μm), TUNEL (scale bar: 50 μm) and cTnT (scale bar: 50 μm) staining of rats in different groups. (C-F) EF, FS, LVIDd and LVIDs calculated. Quantitative analysis of masson (G) and TUNEL staining (H) after different treatments. Representative images of TTC staining (J) and quantitative analysis (K) in different groups. (K-L) Level of inflammatory cytokines (IL-10 and IL-1β) in plasma of MI/RI rats after different treatments. (M) Representative images of major organs by H&E staining (scale bar: 200 μm). Hematological (N) and biochemical analyses (O) after different treatments. Data were presented as mean ± SD, n = 5. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, no significance