Biomimetic astragaloside-IV-loaded mesoporous silica nanoparticles for treatment of dilated cardiomyopathy

Figures

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  FIGURE 1. Representative transmission electron microscopy images of (A) MSNs (scale bar = 200 nm), (B) AS@MSN (scale bar = 500 nm), and (C) AS@MSN/TA/MM (scale bar = 200 nm). (D) Sizes and (E) zeta potentials of the prepared particles. Particle sizes and polydispersity index (PDI) values of AS@MSN/TA/MM after incubation with (F) phosphate-buffered saline (PBS) for 7 d at 4 °C (n = 3) and (G) 10% fetal bovine serum (FBS) for 3 d at 37 °C (n = 3).

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  FIGURE 2. (A) Viability of DOX-induced H9C2 cells incubated with different treatment samples at various concentrations for 24 h. (B) Labeling of live and dead cells by calcein AM/propidium iodide (PI) for different treatment samples (scale bar = 100 μm). (C) Fluorescent images of cells with different treatments and staining with ROS-fluorescent probe DCFH-DA (scale bar = 100 μm). (D) Fluorescence intensity quantification by DCFH-DA vs. AS@MSN/TA/MM, # p < 0.05, ## p < 0.01, ### p < 0.001.

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  FIGURE 3. (A) Representative echocardiographic images. (B) Heart weight index values of the mice after treatment (n = 8). (C) Heart weight to tibia length (HW/TL) ratios of the mice after treatment (n = 8). HW, heart weight; BW, bodyweight; TL, tibia length. For PBS, *p < 0.05, **p < 0.01, ***p < 0.001; for AS@MSN/TA/MM, ## p < 0.01, ### p < 0.001.

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  FIGURE 4. (A) Representative images of H&E staining (scale bar = 50 μm). (B) Representative images of Masson’s trichrome staining (scale bar = 50 μm). (C) Fibrotic area quantification analysis for Masson’s staining (n = 8). (D) Representative images of WGA staining (scale bar = 50 μm). (E) Cross-sectional area quantification for WGA staining (n = 8). H&E, hematoxylin and eosin; WGA, wheat germ agglutinin. The data are presented as mean ± SD vs. PBS, **p < 0.01, ***p < 0.001; vs. AS@MSN/TA/MM, # p < 0.05, ## p < 0.01, ### p < 0.001.

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  FIGURE 5. Serum levels of (A) aldosterone (ALD), (B) angiotensin II (Ang II), and (C) suppression of tumorigenicity 2 (ST2) (n = 8). The data are presented as mean ± SD vs. PBS, *p < 0.05, **p < 0.01; vs. AS@MSN/TA/MM, # p < 0.05, ## p < 0.01.

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  FIGURE 6. (A) Live imaging of dilated cardiomyopathy (DCM) mice and in vivo distribution after treatments with ICG@MSN/TA and ICG@MSN/TA/MM (n = 3 each). (B) Indocyanine green (ICG) accumulated in the heart 12 h after treatment with ICG@MSN/TA (left) and ICG@MSN/TA/MM (right) (n = 3). (C) Quantification of ICG fluorescence intensity from (B), *p < 0.05.

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  FIGURE 7. (A) Hemolysis ratios of the different treatment samples (n = 3); (inset: hemolytic images). (B) Viability of the H9C2 cells after incubation with different treatment samples for 24 h.