Beneficial Roles of Cellulose Patch-Mediated Cell Therapy in Myocardial Infarction: A Preclinical Study

Abstract

Biological scaffolds have become an attractive approach for repairing the infarcted myocardium and have been shown to facilitate constructive remodeling in injured tissues. This study aimed to investigate the possible utilization of bacterial cellulose (BC) membrane patches containing cocultured cells to limit myocardial postinfarction pathology. Myocardial infarction (MI) was induced by ligating the left anterior descending coronary artery in 45 Wistar rats, and patches with or without cells were attached to the hearts. After one week, the animals underwent echocardiography to assess for ejection fraction and left ventricular end-diastolic and end-systolic volumes. Following patch formation, the cocultured cells retained viability of >90% over 14 days in culture. The patch was applied to the myocardial surface of the infarcted area after staying 14 days in culture. Interestingly, the BC membrane without cellular treatment showed higher preservation of cardiac dimensions; however, we did not observe improvement in the left ventricular ejection fraction of this group compared to coculture-treated membranes. Our results demonstrated an important role for BC in supporting cells known to produce cardioprotective soluble factors and may thus provide effective future therapeutic outcomes for patients suffering from ischemic heart disease.

Keywords: bacterial cellulose patch; cell therapy; delivery; implant; myocardial infarction.

Figure 1

Experimental design: acute myocardial infarction (MI), distinct time point (D), echocardiography (Echo), and transplantation (TX).

Figure 2

Placement of the cellulose patch on the left ventricle.

Figure 3

(a). Cell proliferation analysis during 10 days of cell culture on the cellulose membrane. (b). MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) analysis showing the standard curve for cell proliferation. (c). Cell adhesion rate, represented by mean ± SD. The control group mean was 95.17% ± 2.44, whereas the membrane group mean was 87.07% ± 7.77. Control group: polystyrene flask culture. Membrane: cellulose membrane.

Figure 4

(a). SEM analysis showing bacterial cellulose (BC) membrane without cells. Images 500×, scale bar = 50 μm. (b). SEM analysis of the BC membrane containing cocultured skeletal muscle cells and bone marrow mesenchymal stem cells (for 14 days). Arrows indicate cell adhesion. Images 350×, scale bar = 50 μm.

Figure 5

Graphs show echocardiographic measurements of left ventricular end-diastolic (LVED). Mean values were calculated for each group seven days (D7) after and 30 (D30) days after MI. Group I: MI without treatment as a control group; group II: implantation of the BC matrix on the left ventricular surface; and group III: Implantation of the BC matrix seeded with cocultured cells on the left ventricular surface. Standard Deviation: SD, Ns: not significant, asterisk: significant.

Figure 6

Graphs show echocardiographic measurements of left ventricular end-systolic (LVES). Mean values were calculated for each group seven days (D7) after and 30 (D30) days after MI. Group I: MI without treatment as a control group; group II: implantation of the BC matrix on the left ventricular surface; and group III: Implantation of the BC matrix seeded with cocultured cells on left ventricular surface. ns: not significant. Asterisk: significant.

Figure 7

(a). Immunostaining of the post-transplantation cardiac scar tissue. Sections were stained with anti-BrdU antibodies after 30 days of infarction without cells (group II). The arrows point to the cellulose membrane implanted as a patch; 400× (optical light microscopy). (b). Cocultured cells grafted in the post-transplantation cardiac scar tissue. Sections were stained with anti-BrdU antibodies (dark) after 30 days of infarction (group III). The arrows indicate engrafted cocultured cells and vessels; 400× (optical light microscopy).