Functional and Histological Analysis of Stem Cell and Amniotic Membrane Implantation After Acute Myocardial Infarction with Left Ventricular Dysfunction: Experimental Study

Figures

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  Figure 1. Box-plot representing the analysis of the variables Gomori Trichrome (A), Collagen type I (B) and III (C) in Control, SC, HAM and SC + HAM groups. Legend: HAM: Human Amniotic Membrane; SC: Stem Cells.

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  Figure 2. Gomori trichrome staining of myocardial tissue in Control, SC, HAM, and SC + HAM groups. (A) Control: extensive fibrosis (blue) in the infarct region. (B) SC: moderate fibrosis reduction. (C) HAM: further reduction in collagen deposition. (D) SC + HAM: minimal fibrosis and better preserved myocardium. Magnification 400×; scale bar = 25 µm. Blue: collagen/fibrosis; red: myocardium.

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  Figure 3. Visualization of type I collagen (in red—pointed by the red arrow) and type III collagen (in green—blue arrow) in optical microscopy of myocardial tissue slide using Sirius-red staining, in Control, SC, HAM and SC + HAM groups, respectively. The images were obtained from the infarction zone. Magnification 400×; white scale bar = 25 µm. Legend: HAM: Human Amniotic Membrane; SC: Stem Cells.

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  Figure 4. Box-plot representing the analysis of the variables α-actin, CD31, CD68, desmin and sarcomeric actin between the study groups, in Control, SC, HAM and SC + HAM groups. Legend: HAM: amniotic membrane; SC: Stem Cells; *: outlier.

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  Figure 5. Visualization of the α-actin marker (white arrow) on a myocardial tissue slide under light microscopy, in Control, SC, HAM and SC + HAM groups, respectively. The images were obtained from the infarction zone. Magnification: 200×. Scale bar: 50 μm. Legend: HAM: Human Amniotic Membrane; SC: Stem Cells.

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  Figure 6. Visualization of the sarcomeric actin marker (red arrow) on a myocardial tissue slide under light microscopy, in Control, SC, HAM and SC + HAM groups, respectively. The images were obtained from the infarction zone. Magnification: 400×. White scale bar: 25 μm. Legend: HAM: Human Amniotic Membrane; SC: Stem Cells.

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  Figure 7. Flowchart of animal inclusion, exclusions, and mortality throughout the experimental protocol.

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  Figure 8. Flow cytometry histograms showing the expression of surface markers CD45, CD34, and CD90 in bone marrow mononuclear cells. CD45+ cells indicate hematopoietic lineage; CD34+ cells represent hematopoietic stem and progenitor cells; and CD90+ cells suggest the presence of mesenchymal stem cells. The magenta peaks represent marker-positive populations, while the green peaks represent marker-negative populations.

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  Figure 9. (A) Lateral thoracotomy for myocardial access in an animal model; (B) myocardial ischemia resulting from ligation of the left coronary artery (white arrow).

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  Figure 10. Median Sternotomy. (A) Implantation of stem cells (SC) in the myocardium of an animal model (white arrow). (B) Human amniotic membrane implant (HAM) in the myocardium of an animal model (white arrow).

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  Figure 11. (A) Examiner performing an echocardiogram on one of the animals. (B) View of the Systolic Volume obtained by echocardiogram from one of the animals. (C) View of the Diastolic Volume obtained by echocardiogram from one of the animals.