Treatment of myocardial infarction via sodium alginate microsphere-mediated co-delivery of Corydalis yanhusuo ethanolic extract and mesenchymal stem cells

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  Fig. 1. Characterization of the Morphology, Structure, and Physicochemical Properties of Sodium Alginate Microspheres, and the Preparation and Release Behavior of CYEE. A Schematic of microsphere preparation via gas-shearing. B Bright-field and fluorescence images of homogeneous sodium alginate microspheres. C Particle size distribution analysis of sodium alginate microspheres. D Bright-field images of microspheres of different sizes. E Relationship between airflow velocity and sodium alginate microspheres across different size ranges. F Scanning electron microscopy (SEM) images of sodium alginate microspheres. G-J Mechanical and physicochemical property tests of sodium alginate microspheres, including stress–strain curves (G), Young’s modulus analysis (H), swelling behavior (I), and in vitro degradation behavior (J). K Schematic of CYEE release. L In vitro release curve of CYEE from sodium alginate microspheres. Statistical data are expressed as mean ± SD (n = 3)

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  Fig. 2. Biocompatibility Evaluation of the Sodium Alginate Microsphere Co-loading System. A Schematic diagram of sodium alginate microspheres co-cultured with H9C2 cells. B Live/dead staining images of H9C2 cardiomyocytes co-cultured with sodium alginate microspheres for 1, 4, and 7 d; the control group (Con) had no microspheres. C Live/dead staining images of CYEE and MSCs co-loaded within sodium alginate microspheres. D Quantitative statistical analysis of the corresponding live/dead staining results. E Quantitative analysis of MSCs viability in the co-loading system. F CCK-8 cell viability assay results for H9C2 cells treated with different concentrations of CYEE (0, 10, 20, 40, 80, and 160 μg/mL). G Cell proliferation capacity (CCK-8) of the co-loaded MSCs. H F-actin staining images of MSCs under different treatment conditions (Con, LPS, LPS + CYEE, LPS + MSCs, LPS + MSCs + CYEE). Statistical data are expressed as mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA; significant differences are marked as ****p < 0.001

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  Fig. 3. Promotion of in Vitro Migration and Angiogenesis by the Sodium Alginate Delivery System. A Representative images of the endothelial cell scratch healing assay at 0, 12, and 24 h for different treatment groups. B Quantitative analysis of cell migration rate at 12 h. C Quantitative analysis of cell migration rate at 24 h. D Representative fluorescence images of the endothelial cell tube formation assay. E Quantitative statistical analysis of the number of nodes in the tube formation assay. F Quantitative statistical analysis of the total tube length in the tube formation assay. G Live/dead staining images for the evaluation of MSCs inflammation tolerance under LPS-induced inflammatory conditions. H-J Detection of TNF-α, IL-6, and IL-1β expression levels in cardiomyocytes under different concentrations of LPS induction in the in vitro anti-inflammatory capacity evaluation. Statistical data are expressed as mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA; significant differences are marked as *p < 0.05, **p < 0.01, and ***p < 0.001, ****p < 0.0001

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  Fig. 4. Evaluation of in Vivo Therapeutic Efficacy in a Rat Myocardial Infarction Model. A Schematic diagram showing the experimental workflow for myocardial infarction model construction, therapeutic intervention, echocardiographic evaluation, and tissue sampling. B Representative echocardiographic images of the Control group (Sham), myocardial infarction group (MI), CYEE + Gel group, MSCs + Gel group, and CYEE + MSCs + Gel group at 7 and 28 d after the surgery. C-F Quantitative statistical analysis of EF, FS, LVIDd, and LVIDs. G H&E staining images of major organs (liver, spleen, lung, and kidney). Statistical data are expressed as mean ± SD (n = 4). Statistical analysis was performed using one-way ANOVA; significant differences are marked as *p < 0.05, **p < 0.01, and ***p < 0.001, ****p < 0.0001

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  Fig. 5. Histological and Immunofluorescence Analysis of Myocardial Repair and Remodeling. A H&E and Masson’s staining images of heart tissue sections from different treatment groups. B Magnified images of the infarcted myocardial area in Masson’s staining. C Quantitative analysis of left ventricular wall thickness in the infarcted area. D Quantitative analysis of the infarct area percentage. E Representative immunofluorescence staining images of heart tissue showing the expression of cTnT, α-SMA, and Ki67, and their merged images with DAPI. F–H Quantitative statistical analysis of the fluorescence intensities of cTnT, α-SMA, and Ki67. Statistical data are expressed as mean ± SD (n = 4). Statistical analysis was performed using one-way ANOVA; significant differences are marked as *p < 0.05, **p < 0.01, and ***p < 0.001, ****p < 0.0001