Injectable selenium-containing polymeric hydrogel formulation for effective treatment of myocardial infarction

Figures

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  FIGURE 1. Synthesis and characterization of Se-PEG-PPG. (A) Synthesis pathway of Se-PEG-PPG; (B) 1HNMR spectrum of Se-PEG-PPG with detailed peak distribution; (C) FT-IR spectrum of Se-PEG-PPG.

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  FIGURE 2. In vitro characterization of Se-PEG-PPG. (A) H9C2 cell viability after Se-PEG-PPG treatment; (B) Antioxidant activity of Se-PEG-PPG hydrogel assessed using DPPH; (C) The free radical scavenging rate of different concentrations Se-PEG-PPG measured at λ max of 520 nm using DPPH; (D) Rheological properties of 2% Se-PEG-PPG; (E) Rheological properties of 3% Se-PEG-PPG. Black and red represent storage modulus (G′) and loss modulus (G″), respectively; (F) In vitro gelation of Se-PEG-PPG hydrogel at 37°C and SEM images of Se-PEG-PPG hydrogel before and after gelation. Scale bar, 10 μm.

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  FIGURE 3. Representative sectioning and imaging of myocardium after hydrogel injection. Blue (DAPI), cell nucleus; green (FITC), Se-PEG-PPG hydrogel; Scale bar, 100 μm.

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  FIGURE 4. The performance of cardiac function post MI. (A) Myocardial function has improved significantly at 28th day post MI; (B,C) Quantitative analysis of left ventricular ejection fraction (EF) and shortened fraction (FS) on the third day after MI (n = 5, ***p < 0.01); (D,E) Quantitative analysis of left ventricular ejection fraction (EF) and shortened fraction (FS) at 28th day post MI (n = 5, ***p < 0.01); (F,G) Quantitative analysis of left ventricular diastolic inner diameter (LVIDd) and left ventricular systolic inner diameter (LVIDS) at 28th day post MI (n = 5, ***p < 0.01); (H) Representative echocardiographic images on the third day after MI; (I) Representative echocardiographic images at 28th day post MI.

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  FIGURE 5. Expression of inflammatory cytokines in serum on the third day after MI. (A) Diagrammatic sketch of downregulation of IL-6 expression after intramyocardial injection with Se-PEG-PPG hydrogel; (B) Expression level of IL-6 (n = 4, ***p < 0.01); (C) Expression level of IL-1β (n = 5, ***p < 0.01); (D) Expression level of TNF-α (n = 5, ***p < 0.01).

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  FIGURE 6. Poly (DH-Se/PEG/PPG urethane) attenuates cardiac fibrosis MI induced. (A) Schematic illustration of decreased fibrosis and fibrosis-related makers after injection of Se-PEG-PPG hydrogel; (B) Masson’s trichrome staining of heart was shown. Collagen fibers were stained blue and myocardium was stained red. Scale bar: 10 mm; (C) Relative mRNA expression levels of α-SMA, Collagen Ⅰ and Collagen Ⅲ were estimated by RT-PCR and normalized to those of GAPDH (n = 5, ***p < 0.01); (D,E) The protein expression level of α-SMA, Collagen Ⅰ and Collagen Ⅲ in mouse cardiac tissues (n = 6, ***p < 0.01).